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Accurate c-myc-olfactory receptor quantification oncharacterized natural nanovesicles for biosensor

applicationsMarta Sanmarti-Espinal, Patrizia Iavicoli, R Galve, Marie-Annick Persuy,

Edith Pajot-Augy, M.P. Marco, J. Samitier

To cite this version:Marta Sanmarti-Espinal, Patrizia Iavicoli, R Galve, Marie-Annick Persuy, Edith Pajot-Augy, et al..Accurate c-myc-olfactory receptor quantification on characterized natural nanovesicles for biosensorapplications. International Conference on Nanotechnology in Medicine - Nanomed, Nov 2012, Londres,United Kingdom. pp.2. �hal-02747921�

University College London (UCL), UK

Abstract Book 7-9 November 2012

NANO-1 Multi-layer Application of NanoMed close to the market

M. Graf zu Eulenburg, InovisCoat GmbH, Rheinparkallee 3, D-40789 Monheim.

The demand on the functional properties and surface characteristics of medical devices has increased dramatically. Therefore the market is driven by the development of new multifunctional medical products with contents of nano-scaled particles. In the near future new products with new surface properties will control the market, whether we look at the diagnostic or the pharmaceutical markets. To produce such multifunctional and “surface-enhanced” products, one has to focus on the complex application of very thin and ultra-precise layers onto a substrate. This publication will show new trends in research and development as well as mass production of coating technologies useful for the application of one and especially multi-layer systems between 100 nm and 10 µm. The production of coated products is a very versatile process. A large number of influences are determining the final product quality, like preparation of substrates, coating and drying process, atmosphere conditions etc. This oral speech will focus on the continuous application of wet coating layers in a roll to roll process. The advantages of this production process are the relatively easy application and low level of investment needed in the machinery equipment. Generally speaking the coating method is influenced by the substrate, the viscosity and the solid content of the coating chemistry and the requested coating layer thickness. Depending on the evaluation of all named criteria it is possible to recommend a method. Nevertheless it is necessary to do test runs for an optimized selection of the right coating method. The coating systems described in this talk are: slot die technology, ultrasonic spray technology, micro roller application, dipping system and multilayer cascade coating.

NANO-2 Lysine amino acid and gold nanoparticles functionalized mesoporous silica materials synthesis and their application in gene delivery H. K. Daima, PR. Selvakannan, S. Shankar, S. Bhargava, V. Bansal Centre for Advanced Materials and Industrial Chemistry (CAMIC), School of Applied Sciences, College of Science, Engineering and Health, RMIT University, Melbourne (VIC) 3000 Australia

Mesoporous silica materials have been extensively used as catalysts supports, adsorbents and drug/gene delivery. In the present work, we have synthesized different kinds of mesoporous silica materials functionalized by lysine amino acid and gold nanoparticles via the modified triblock copolymer P123 template method and using them as non-viral DNA delivery vehicles. All these materials have very high surface area, tuneable pore size, high pre volume and different surface functionality in order to enhance the adsorption of plasmid DNA and their transformation. Subsequently, these materials were screened for their application in gene delivery by transferring ampicillin resistance gene and green fluorescence protein (GFP) gene containing plasmid DNA (pDNA) in to model microorganism E. coli DH 5α. At ambient temperature, different weight ratios of pDNA and silica nanomaterials were physically mixed for DNA adsorption and we have observed that at 1:5 ratios, the transformation efficiency reached its maximum. Adsorption of DNA within the nanochannels of mesoporous materials was proved by the lack of mobility of DNA observed in the agarose gel electrophoresis in mobility gel assay. Since DNA adsorbed within the pores, the DNA can be protected form enzymatic digestion. Gene expression of green fluorescence protein (GFP) gene was shown in the cellular environment using E. coli. Among the different materials, gold nanoparticles functionalized mesoporous silica materials were found to be most efficient to deliver gene due to the high affinity of gold nanoparticles to the DNA and their larger pore size. In all

these cases, DNA was adsorbed within these pores in a super coiled form, which retains their structural integrity and functionality. NANO-4 In-situ Re-endothelialization via Multifunctional Nano-scaffolds L. C. Su1, H. Xu2, Y. T. Tsai1, L. Tang1, J. Yang1, K. T. Nguyen1

1University of Texas, Arlington, Texas, USA. 2University of Texas Southwestern Medical Center, Dallas, Texas, USA.

Endothelium monolayer lining blood vessels consists of anti-thrombotic activities; however, damages to this layer will expose highly thrombogenic subendothelium and stimulate the formation of thrombosis [1]. In order to regenerate endothelium monolayer on the damaged vessel wall in situ, novel urethane-doped polyester (UPE) nanoparticle scaffold system with dual ligands, injured artery-targeting ligand (e.g. GPIb ligands) and endothelial progenitor cell (EPC)-binding molecule (e.g. anti-CD34 antibodies), has been developed. These nanoparticle scaffolds can target the sites of damaged endothelium/injured arterial wall and further prevent platelet deposition via GPIb ligands as well as recruit endothelial progenitor cells (EPC) from the blood stream through anti-CD34 antibodies. These nanoparticle scaffolds provide an optimized substrate to maturate EPCs on the top of exposed sub-endothelial lumen by controlled release of growth factors including vascular endothelium growth factor (VEGF). The spherical UPE scaffolds ranged 400nm in average diameters were fabricated using a precipitation method. These nanoparticle scaffolds were stable, biocompatible, and hemocompatible. Both in vitro and ex vivo targeting performance of conjugated nanoscaffolds not only showed improved binding efficiency comparing to controls, but also possessed competitive bindings to platelets. In vivo animal studies also revealed the cell capturing and platelet mimicking functionalities of our nano-scaffolds [1] A.B. Baker, W. J. Gibson, V. B. Kolachalama, M. Golomb, L. Indolfi, C. Spruell, E. Zcharia, I. Vlodavsky, E. R. Edelman, J. Am Coll

Cardiol., 59, 1551 (2012). [2] Y. A. Naimushin, A. V. Mazurov, Platelets, 15, 419 (2005).

NANO-5 Combined Targeted Drug Delivery and Mild Hyperthermia Using Magnetic Nanocomplexes for the Targeted Treatment of Cancer T. Mitrelias1,2,*, M. Tselepi1,2, A. Romanov3, V. Orel3

1Cavendish NanoTherapeutics Ltd, CPC1 Capital Park, Fulbourn, Cambridge, CB21 5XE, UK. 2Cavendish Laboratory, University of Cambridge, Cambridge, CB3 0HE, UK. 3 National Cancer Institute, Kiev, Ukraine.

Cancer remains the second leading cause of deaths worldwide. Technologies that would allow targeted destruction of tumours by localising the delivery of therapeutic agents to the tumour region are very promising to assist in the battle against cancer. We are developing systems which utilise magnetic nanoparticles loaded with therapeutic agents. The cancer cells are killed by the combined action of the drugs and a local temperature increase in the area around the tumour site using externally applied electromagnetic irradiation (hyperthermia). Our preliminary clinical trials and pilot studies have demonstrated significant effects in the reduction of the size of a tumour and a 50% to 300% increase in the survival rates (on average) of patients with numerous cancers. This paper examines the effects of nano-complexes formed by iron oxide magnetic nanoparticles and doxorubicine (DOXO) on nonlinear growth dynamics of Lewis lung carcinomas. One of the ways this can be

achieved is by making magnetic nanoparticles which are able to resonantly respond to a time-varying electromagnetic field; thereby energy can be transferred from the excitation field to the nanoparticles and subsequently to the target area. Our study has shown that the growth factor for tumour volumes on the 12th and 17th day after tumour transplantation to mice was maximum after treatment by the nano-complexes and electromagnetic excitation. A most significant anti-metastatic tendency was also demonstrated following treatment with our approach. Following our patented synthesis procedure, the nano-complexes show higher magnetisation, but less agglomeration, as well as a higher concentration of paramagnetic centers which is directly related to higher concentration of free radicals which could be responsible for the observed effects. NANO-6 Tadalafil Nanocrystals: A Novel Nanofabrication Approach for Solubility and Dissolution Rate Enhancement M.M. Mehanna, A.N. Allam Faculty of Pharmacy, Alexandria University, Alexandria, Egypt

Tadalafil is used to treat erectile dysfunction. Tadalafil is characterized by low solubility and high permeability (a class II drug), which has a negative influence on its bioavailability. Utilization of nanocrystals formation approach represents an effective and facile methodology to enhance the poorly soluble drugs dissolution rates. Our aim was to select and optimize various parameters that affect the preparation of tadalafil nanocrystals, which were prepared by the antisolvent precipitation method. Preliminary studies were carried using Pluronic F-127, Pluronic F-68, Hydroxypropyl methylcellulose (50 and 4000 c.p.), polyvinyl chloride, and polyvinyl alcohol as stabilizers. In order to shed more lights on the optimum conditions leading to the preparation of tadalafil nanocrystals, the effects of the studied parameters on crystal size, solubility and dissolution rate were investigated. The results obtained pointed out that the use of methanolic solution of tadalafil to water as the antisolvent produced crystals in the submicron range in presence of a stabilizer. The particle size decreased with the increase of the stabilizer concentration but further increase of which didn't affect the crystals size significantly. IR spectroscopical studies revealed that there was no chemical changes in tadalafil structure occurred during nanofabrication. The differential scanning calorimetry (DSC) analysis illustrated that there was no substantial crystalline change in the nanocrystals compared to the raw drug. Both the solubility and the in-vitro dissolution rate of tadalafil were significantly augmented through introduction of the nanocrystals. NANO-7 Nanocapsules Based Pediatric Liquid Lisinopril Formulation: Design and Physicochemical Characterization M. M. Mehanna Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.

Lisinopril is an orally active angiotensin-converting enzyme (ACE) inhibitor used for the treatment of hypertension, heart failure, and acute myocardial infarction in infants. It is currently supplied as tablets. Solid dosage forms represent a problem in children with swallow difficulty and in neonates. There is no commercially available oral liquid preparation of lisinopril. Our strategy is to prepare oral liquid formulation of lisinopril based on lipid-core nanocapsules. Nanocapsules were prepared according to the nanoprecipitation protocol utilizing poly-ε-caprolactone as a shell-forming polymer. The following characteristics of nanocapsules were determined: encapsulation efficiency, nanocapsules size, in-vitro drug release and stability on storage. The results revealed that the optimized formulations of lisinopril-loaded nanocapsules were in the nanometric range with a high encapsulation efficiency (more than 90%), both stable for 3 months. The release of lisinopril from the polymer-

based nanocapsules was rapid in the simulated gastric fluid. In the top of that, the concentration of lisinopril in the liquid preparation was appropriate for oral pediatric administration. NANO-9 An integrated solution for rapid end-user ready biosensing Yongbai Yin, David.R. McKenzie and Marcela .M.M. Bilek

Biomedical Materials Group, School of Physics, University of Sydney, NSW 2006, Australia

A novel integrated biosensor methodology is proposed and demonstrated. The methodology utilizes a nitrogen-containing plasma polymer to achieve linker-free binding of biorecognition molecules that allows the sensor surface to be freeze dried for long shelf life under ambient conditions. The sensor is configured for single wavelength ellipsometric detection providing a low cost, versatile, and rapid sensing and diagnosis platform suitable for a wide range of applications and end-users. The merits of the methodology were demonstrated using three antigen-antibody pairs, which showed the desirable features of long shelf life, simple storage procedure, reliable performance and minimal involvement of end-users. The covalent binding mechanism of the NPP surface was investigated. NANO-10 Potentiality of ORMOSIL nanoparticles as negative Contrast Agents for MRI M.C. Gonçalves1, G. Feio,2, A.Carvalho2

1Dep. Chemical Eng., IST, UTL, Av. Rovisco Pais, 1049-001 Lisboa, Portugal, and ICEMS, Av. Rovisco Pais, Portugal 2CENIMAT-I3N/DCM/FCT/UNL, Campus da Caparica, 2829-516 Caparica, Portugal

We present a new application for ORMOSIL as contrast enhancement agents for magnetic resonance imaging [1]. For long that these materials are use in nanomedice mainly for gene or drug deliver, some applications in other imaging modalities, as fluorescence have been reported, but as far as we know as negative contrast agent for IRM this is a new approach. We measured the relaxivities r1, r2 and r2

* for different ORMOSIL and silica [2] with a superparamagnetic core of iron oxide nanoparticles of 6nm diameter. The measurement of the longitudinal and transversal relaxation times was obtained in a Bruker AVANCE III spectrometer in a magnetic field of 7 T (300 MHz for proton). The preliminary results allows to say that ORMOSIL effect on T1 is minor but on T2 and mainly on T2

* is is a huge effect, this behaviour is the good one, for a potential negative contrast agent. [1] A.E. Merbach and E. Tóth in The Chemistry of Contrast Agents in Medical Magnetic Resonance Imaging (Wiley, 2001) [2] C. Huanga, K. G. Neoha , L.Wanga, E. Kanga and B. Shuter, Contrast Med.Mol. Imaging, 6, 298 (2011)

NANO-11 New long circulating magnetoliposomes as negative contrast agents for MRI M.B. Martins1, L. Corvo1, P. Marcelino2, F. Fontes3,4, H.S. Marinho4, G. Feio,3, A.Carvalho3 1iMed/FF/UL, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal 2CEDOC/ FCM/ UNL, Campo dos Mártires da Pátria, 1169-056 Lisboa, Portugal 3CENIMAT-I3N/DCM/FCT/UNL, Campus da Caparica, 2829-516 Caparica, Portugal 4DQB & CQB, FC/UL, Campo Grande, 1749-016 Lisboa, Portugal

For a long time liposomes and in particular magnetoliposomes have been used in medicine due to their remarkable properties, namely: a) high biocompatibility, b) possibility of several surface manipulations, c) possibility to enclose both hydrophilic as well as hydrophobic compounds, d) ability to simultaneously contain imaging and therapeutic agents and c) controllable size, from 20 nm to 1 µm with a relatively high degree of accuracy. We present in this work the potentiality of a new long circulating liposomal formulation coated with polyethylene glycol (PEG) loaded with PEG-coated 10 nm iron oxide nanoparticles, as a negative contrast agent for MRI. The encapsulation parameters obtained and the size of the loaded liposomes make this system a good tool to be used as a negative contrast to visualize inflammatory sites after ischemia-reperfusion injuries. The described magnetoliposomes were tested in a rat model of liver ischemia, performed after hepatic artery and porta vein occlusion. The relaxivities r1, r2 and r2

* were obtained in a Bruker AVANCE III spectrometer in a magnetic field of 7 T (300 MHz for proton), a very interesting value for the clinical applications. MRI images, at 7 T, were obtained for rat liver after 30 min of partial ischemia and 24 h of post-treatment with and without the developed magnetoliposomes, and the contrast enhancement measured. The long circulating nanosystem developed by us had a minor effect on T1 but on T2 and mainly on T2

* it has a huge effect. Therefore the results obtained by this preliminary study indicate that this nanosystem may be a potential negative contrast agent. Financial support: PEst-OE/QUI/UI0612/2011, Pest-OE/SAU/UI4013/2011, Sociedade Portuguesa de Transplantação - Bolsa de Investigação SPT/ Astellas Strategic Project Pest-C/CTM/LA0025/2011 Portuguese Nuclear Magnetic Resonance Network (PTNMR) NANO-12 Production of electrospun cellulose acetate fiber mats as carriers of a natural oil from Copaifera sp. A.L.G. Millas1, J. V. W. Silveira1, E.F. Tessaroli1, E. Bittencourt1

1State University of Campinas, Cid. Universitária, Campinas/SP, 13083-856, BR.

The science of biomaterials and nanotechnology combine in the search of new alternatives to improve the physical, chemical, and mechanical properties of materials associated with the high surface area to volume ratio, and nanometer dimensions, that provide optimum performance with small amounts of the nano-material. Copaiba (Copaifera langsdorffii) natural oil is known for its wound healing, analgesics, antifungal and antimicrobial functions. Biodegradable and biocompatible ultrafine fibrous webs of cellulose acetate (CA) containing copaiba oil were prepared by electrospinning and tested for their main properties. The effect of CA and oil concentrations on fiber morphology was elucidated. Cellulose acetate (CA) at 10 (%w/w) was dissolved in acetone and water system (4:1) with differents concentrations of oil, between 1% and 15%. GC was used to identify the components while SEM and TGA data revealed the influence of the oil phase on the morphological and thermal characteristics of the obtained electrospun fibers. The changes in viscosity and evaporation rate of precursor solutions formulated with copaiba allowed the formulation of defect-free fibers while the thermal properties were not significantly affected. Tests of cell viability in vitro were performed. The obtained nanofibers are expected to be useful in applications in the regenerative medicine field, as bandages and scaffolds.

NANO-13 Nanoscale plasma treatment for improvement behavior of F-799 alloys for MoM prosthesis application. C. Diaz1, J.A. García1, S. Mändl2, R. Pereiro3, B. Fernández3

1AIN - Centro de Ingeniería Avanzada de Superficies, 31191 Cordovilla-Pamplona, Spain 2Leibniz-Institut für Oberflächenmodifizierung, 04318 Leipzig, Germany 3Universidad de Oviedo, Facultad de Química, Avda. Julián Clavería, 8,33006 Oviedo, Spain

Metal-on-metal hip (MoM) replacement systems still have risks of revision as a consequence of possible high wear rates, low corrosion resistance or release of toxic ions [1]. Because the metal head and the metal cup slide against each other during walking or running, some tiny metal particles may wear off of the device and enter into the space around the implant, leading to wear-particle induced osteolysis [2]. Therefore, some of the metal ions from the metal implant or from the metal particles may even get into the bloodstream [3]. For these replacement systems the most used metal materials are Ti and CoCr alloys: CoCr alloys are preferred in total joint replacements, in both supportive and articulating locations due to their higher wear resistance [4]. However, these alloys do exhibit lower corrosion resistance than Ti alloys [5]. The wear resistance of Ti and its alloys is relatively low and these materials may not be used where contact wear can occur. As an important consequence, metallosis can be induced by the release of wear metal particles, is often a consequence of wear due to metal-to-metal contact [6]. Hence, a novel nanoscale surface modification of CoCrMo alloy (F-799) is presented for obtaining improved surface conditions in the CoCr alloys for these applications. The aim is to improve corrosion resistant of these alloys, in order to reduce the released ions as much as possible from surface of this material as a consequence of its lower wear rate and reduction in corrosion effects. For that, three different treatments at temperatures of 300, 350 and 400ºC were carried out by plasma immersion ion implantation technique (PIII). Results show an improvement in corrosion resistance of CoCrMo alloys by introducing oxygen in relation to untreated reference material. Wear rate studies showed very similar behaviour after the treatments and XPS analysis were carry out in order to know what modifications in the structure, oxygen was introducing after each treatment. As consequence of these results, ion release rate from the treated surface of CoCrMo alloys was reduced in comparison with untreated CoCrMo alloy. [1] U. Holzwarth, G. Cotogno, JRC Scientific and policy reports, European Comission, (2012). [2] C: Wedemeyer, J. Xu, C. Neuerburg, S. Landgraeber, N.M. Malayar, F. von Knoch, G. Gosheger, M. von Knoch, F. Loer, G. Saxler,Calcif. Tissue Int. 81, 394-402 (2007). [3] I. Catelas, J. D. Bobyn, J. B. Medley, J. J. Krygier, D. J. Zukor, O. L. Huk, Wiley Periodicals, Inc, (2003). [4] J. Cawley, J.E.P. Metcalf, A.H. Jones, T.J. Bandb, D.S. Skupien, Wear 255, 999–1006, (2003). [5] J.R. Davis, Handbook of Materials for Medical Devices, ASM International, (2003). [6] P. Korovessis, G. Petsinis, M. Repanti, T. Repantis, J Bone Joint Surg Am, 01;88(6):1183-1191, (2006). [7] S. Mändl, B. Rauschenbach, Surface and Coatings Technology, 156, 276–283, (2002). [8] C. Díaz, J. Lutz, S. Mändl, J. A. García, R. Martínez, R. J. Rodríguez, J. J. Damborenea, M. A. Arenas, A. Conde, Phys. Stat. Sol. (c), nº 4, 947-951, (2008). [9] C. Díaz, J.A. García, S. Mändl, R.J. Rodríguez, IEEE Issue - Applications and numerical Simulation of Plasma Surface Modification, (2011). [10] K.G. Kostov, M. Ueda, M. Lepienskyc, P.C. Soares, G.F. Gomes, M.M. Silva, H. Reuther, Surface & Coatings Technology, 186, 204–208, (2004). [11] J.A. García, C. Díaz, S. Mändl, J. Lutz, R. Martínez, R.J. Rodríguez, Surface & Coatings Technology 204, 2928–2932, (2010).

NANO-14 The Phosphatidylinositol Microcapsules for Successful Therapy of Dangerous Diseases A.N. Begzat, Sh.M. Nurmoldin, Kaster, A.A. Yesmambetov, M.K. Gilmanov Scientific Research Institute of the Problems of Biology and Biotechnology of Al-Faraby’s Kazakh National University, Almaty, Al-Faraby avenue 71, Kazakhstan

The delivering of the drug by digestive system and blood circulation system cause the toxic effects on all cells of the organism and led to appearance different pathologies. There is the big necessity to create the new systems for direct delivering of the drugs to the ill organ without participation of the digestive and blood circulation systems. Therefore we developed the new micro carriers from new material – phosphatidylinositol (PI) for direct delivering drugs to ill organ. In contrast from other phospholipids PI related to charged phospholipids. In this reason the microcapsules which made from PI have a negative charge and they are very small – less than 1mkm in diameter. Due to this charge every microcapsule push up each other and they never aggregate. Our PI microcapsules very stable, so they can be stored without any changes in sterile condition several years. Also we developed the very effective method of the loading of the PI microcapsules by drugs. The principle of this method is next. In hydrophobic solution our PI microcapsules is opened like bivalved shell. When we transfer PI microcapsule into hydrophilic solution the PI microcapsules begin to close and its scoops the solution with dissolved drug. Such drugs are located inside of the microcapsule. The PI microcapsules loaded by drugs were tested in several scientific research institutes in Almaty. The loaded PI microcapsules were mixed with ointment and then for therapy ointment with microcapsules is anointed on the skin of the field of the ill organ. It was established that they loaded microcapsules easily path through skin and muscles due to thermal movement. They could not penetrate into the cells of the skin and muscles, because they have the same charge too. Thus, loaded PI microcapsules without hindrance pass through intercellular spaces and reach ill organ during 15 minutes, as it was established by cardiograph. The PI microcapsules loaded by different drugs show success in therapy of the next diseases: glaucoma, macular dystrophy, diabetic food, spinal tuberculosis, sialadenitis and arthritis. The tests were carried out on hundred patients and it was received six patents for therapy of different diseases. NANO-15 Magnetite/Chitosan Biocompatible Nanocomposite as an Effective Anti-Bacterial Agent

A. Sharifian-Esfahani, M.T. Salehi, M. Nasr-Esfahani, and E. Ekramian

Department of Material Science and Eng., Najafabad Branch, Islamic Azad University, Najafabad, Iran

The iron oxide species that has gained the most attention for medical applications is magnetite (Fe3O4)1. Nowadays, they are promising materials for cancer diagnosis and therapy 2,3 ,because the Food and Drug Administration approved that superparamagnetic Fe3O4 nanoparticles are biocompatible with human body4 . On the other hand superparamagnetic iron oxide (SPIO) is particularly attractive as antibacterial agent. Natural polymers such as dextran, chitosan or starch can be attached to the surface of iron oxide nanoparticles in order to minimize the adsorption of proteins and to protect them from macrophages2. In the present work, Fe3O4 magnetic ferrite nanoparticles were fabricated by chemical co-precipitation method5, and were coated with chitosan as suitable biocompatible outer shells during the co-precipitation processing with a uniformly stable monodispersed colloidal ferrofluid. The morphological and magnetic properties of the magnetic nanoparticles were characterized by different techniques (TEM, XRD, VSM, FTIR, etc.). At room temperature, the samples exhibit superparamagnetic behavior. Both naked and chitosan coated SPIO nanoparticles were effective against bacteria. [1] K.B. Hartman, L. J. Wilson and M. G. Rosenblum, Mol. Diag. Ther., 12, 5 (2008).

[2] A. K. Gupta, M.Gupta, Biomaterials, 26, 3995 – 4021 (2005). [3] A. Jordan, R. Scholz, P. Wust, H. Fahling, R. Felix, Journal of Magnetism and Magnetic Materials, 201, 413 – 419 (1999). [4] M. Lattuada and T. A. Hatton, Langmuir 23, 2158 (2007). [5] R. Massart, IEEE Trans. Magn. MAG-17, 1247 (1981).

NANO-16 Tissue Engineering & Biomaterials science Shwan A. Hamad1, Simeon D. Stoyanov2 and Vesselin N. Paunov1 1 Department of Chemistry, The University of Hull, Hull, HU6 7RX, UK. 2 Unilever R&D Vlaardingen, Vlaardingen, 3133 AT, The Netherlands.

We report the fabrication of novel shellac-cells composite microcapsules with programmed release of cells upon change of pH in a narrow range.[1] The microcapsules were prepared from yeast cells as a model for probiotics combined with aqueous solution of ammonium shellac doped with a pH sensitive polyelectrolyte, like carboxymethyl cellulose or polyacrylic acid. The cells dispersions in aqueous ammonium shellac were spray-dried or spray co-precipitated to yield composite shellac-cell microcapsules in which the cells retained their viability even when treated with aqueous solutions of very low pH and subjected to mechanical stress. We demonstrate two types of triggered release of yeast cells from these microcapsules with pH trigger and cell growth trigger and evaluate the microcapsules disintegration rates. Depending on the type of the polyelectrolyte integrated in the shellac microcapsules they can be programmed to give very versatile responses ranging from slow cell release to explosive swelling and disintegration at higher pH or exposure to growth media. We show that the cells retain their viability following their release from the microcapsules into the aqueous solution. We modelled the kinetics of living cell release from the composite shellac microcapsules triggered by: (i) pH change, which dissolves the shell and (ii) the growth of the encapsulated cells, when places in a culture media. For pH triggered release of cells from the composite microcapsules, the rate constant of cell release depends on the swelling/dissolution rate of the shellac matrix and varies with the pH of the aqueous media. Such composite microcapsules could find applications in formulations for protection and delivery of probiotic and other cell cultures with programmed and triggered release of the encapsulated cells in cell implants, including stem cells and live vaccines. [1] S. A. Hamad, S. D. Stoyanov and V.N. Paunov, Soft Matter, 8, 5069 (2012).

NANO-17 Encapsulation of Living Cells into Sporopollenin Microcapsules Shwan A. Hamad1, Simeon D. Stoyanov2 and Vesselin N. Paunov1

1 Department of Chemistry, The University of Hull, Hull, HU6 7RX, UK. 2 Unilever R&D Vlaardingen, Vlaardingen, 3133 AT, The Netherlands.

We demonstrate how living cells can be encapsulated inside sporopollenin microcapsules derived from Lycopodium clavatum.[1] Sporopollenin is a major part of the exine of spores and pollens. It can be found in soils and rock sediments and has been recognised as one of the most extraordinary resistant materials of bioorganic origin.[1] Sporopollenin can be isolated from the pollens by consecutive acidic and basic solvent treatment, which removes the inner cellulose wall but morphologically preserves the exine. Recently, we reported a simple and robust method for loading sporopollenin exine of Lycopodium clavatum with functional inorganic and organic nanoparticles synthesised in situ. This technique uses sporopollenin microcapsules as micro-reactors where a chemical reaction generates a low soluble product inside, which precipitates in the form of

nanoparticles usually much larger than the pores of the sporopollenin membrane.[2] To encapsulate large objects like cells, the sporopollenin particles are compressed into a pellet which forces their trilite scars to open up. Our new method involves exposing a sporopollenin pellet to an aqueous suspension of cells in the presence of a surface active agent which facilitates the capillary suction of the cells suspension inside the compressed sporopollenin and its ‘‘re-inflating’’ and closure of trilite scars. We demonstrate that the viability of the cells is preserved after the encapsulation in the sporopollenin capsules which contain a significant amount of entrapped cells and show biological activity when placed into a culture medium. Since the sporopollenin nanopores allow nutrient transport across the capsule wall, it could be used for controlling the rate of in situ fermentation reactions or as bio-reactors. The encapsulation of living cells inside sporopollenin can be used for many different purposes in the food and pharmaceutical industries, including protection of probiotics in foods and delivery of live vaccines for pharmaceutical applications. [1]S. A. Hamad, S. D. Stoyanov, V.N. Paunov, Soft Matter, 8, 5069 (2012). [2]V. N. Paunov, G. Mackenzie, S.D. Stoyanov, J. Mater. Chem., 17, 609 (2007).

NANO-18 The study of phase transfer of InP/ZnS quantum dots via chemisorption, silanization and polymer coating S. Massadeh1, T.Nann2

1King Abdulla International Medical Research Center, Riyadh, 11426, Kingdom of Saudi Arabia. 2Ian Wark Research Institute, University of South AustraliaMawson Lakes Campus Mawson Lakes, AUSTRALIA.

Colloidal semiconductor Quantum Dots (QDs), are a specific type of nanoparticles that have a size range of 1-10 nm. QDs could be really advantageous when used as alternatives to organic dyes; actually QDs are attractive in the field of biosensors owed to their long-term photostability[1]. However, around 80% of the currently investigated QDs are Cadmium containing QDs; the Cd-based QDs are unsuitable for biological applications, since cadmium is a highly toxic heavy metal and may leak out of QDs. Only recently, scientists started to synthesize cadmium free QDs. Herein, we report a study using non-toxic InP/ZnS. Three different approaches have been applied to transfer the InP/ZnS QDs into aqueous buffers. Herein, we report a complete analytical and characterization profile of each method used for the phase transfer of the InP/ZnS QDs. Size Exclusion Chromatography (SEC), Gel Electrophoresis (GE), and Dynamic Light Scattering (DLS) are the main techniques that have been applied for the analysis of the InP/ZnS QDs. Even though the ligand exchange method has been widely discussed in the literature, the main focus of previous work was the use of CdSe, CdTe and CdS QDs, whereas very little work has been reported on InP/ZnS QDs. Different bifunctional ligands have been used to transfer the InP/ZnS QDs into biological buffers. The use of bifunctional ligands for the phase transfer of QDs is a cheap and practical method that has been used frequently in the literature [2]. The most commonly used bifunctional ligands in the ligand exchange method are the mercaptoalkyl acid ligands, the -SH moiety of the ligands adsorbs on the surface of the QDs and replaces the NH2 bound HDA ligands to form a monolayer of hydrophilic ligands by chemisorption, the advantage of this technique is that the mercaptoalkyl ligands maintain an overall small size of the QDs which is desirable for further biological applications. [1] Y.Wang, J. Zheng, Z. Zhang, C. Yuan, D. Fu, Colloids and Surfaces A: Physicochem.Eng.Aspects,Vol.342,p.102,(2009). [2] W.C. Chan, S.Nie, Science, Vol.281, p.2016 (1998).

NANO-19 Culture Free Bacteria Detection P. Drake1, P.S. Jiang1, H.W. Chang1, S.C. Su1, J. Tanha2, L.L. Tay2, P.L. Chen,3 Y.J Lin1 1 Industrial Technology Reserach Institute, Hsinchu, Taiwan 2 National Research Council, Ottawa, Canada 3 Research Centre for Applied Sciences, Academia Sinica, Taipei, Taiwan

The detection of pathogenic bacteria is of paramount importance in areas such as public health. Current detection methods are typically based on culturing and colony counting with results obtained after several days. More advanced techniques such as those based on PCR can deliver results in 24 hours but still rely on a culturing step for samples with low pathogen concentrations below 104 colony forming units (CFU) per mL. There is a great demand for a system that can deliver rapid pathogen detection without the need for enrichment or culturing steps and a detection limit in the 1 – 100 CFU mL-1 range. Nanoparticle labels were used to detect the bacteria Staphylococcus aureus. Two different labels were employed. These consisted of nanoparticles based on iron-oxide for magnetic trapping and isolation of the bacteria and surface enhanced Raman spectroscopy (SERS) active gold nanoparticles (AuNPs) with a 4-mercaptobenzoic acid surface for the detection and quantification of the bacteria. Both types of nanoparticles were coated with single domain antibodies that selectively bind to protein A on the surface of S. aureus. Transmission electron microscopy (TEM) image analysis showed the iron-oxide nanoparticles to have magnetic core diameters of 20 nm ± 3 nm and the gold nanoparticles to be 51 nm ± 2 nm in diameter. A response curve was obtained displaying a logarithmic dependency between the 1073 cm -1 peak intensity in the SERS spectra of the AuNPs and the concentration of S. aureus cells in various samples. The overall detection limit was estimated to be 1 S. aureus cell mL-1 in less than 10 min without culturing. NANO-20 Nanoparticle Labels for the Detection and Identification of Pathogenic Bacteria through DNA Analysis P. Drake, Y.C. Chen, I. Lehmann, P.S. Jiang Industrial Technology Reserach Institute, Hsinchu, Taiwan

A nanoparticle (NP) system was developed for DNA analysis. The nanoparticles were coated with short chain DNA tags. These were then used to identify a target bacterial DNA sequence. The tags function as primers in a standard PCR reaction with the reverse primers and forward primers on different NPs. On binding to the target DNA the primers can be extended through a PCR reaction. Once extended the NPs are able to hybridize together through the extended DNA chain resulting in the formation of a composite nanostructure. The process of extension and annealing can be repeated as in standard PCR reactions building up larger nanostructures. This structure can be trapped and a signal recorded to quantify the initial concentration of the target DNA sequence. The system offers a new approach for ‘Real-Time PCR’ and the potential to multiplex the readout, detecting several different DNA sequences in a single reaction. The nanoparticles were coated with the DNA tags through a standard EDC reaction and cleaned through precipitation or centrifugation followed by decanting. A test assay was performed by replacing the primers in a standard PCR reaction with the nanoparticle system. A signal was obtained from 10 zeptomoles of target DNA, from the signal to noise ratio the detection limit was estimated to be less than 1 zeptomole. For comparison a PCR assay based on the standard SYB Green method was performed. This used the same primers and target DNA and had a detection limit of 10 attomoles, a 1000 times less sensitive.

NANO-21 Studyof some affecting factors on CNTs characteristicssynthesized using chemicalvapor depositionmethodon MgO substrate A.A. Hosseini1,F. Tabatabaee1, M. Shadfar1

1Department of physics, faculty of sciences, state University of Mazandaran, Babolsar, 47416-95447, Iran.

Carbonnanotubes(CNTs)areallotropes of Carbon whichare formed by rollingone or more graphen sheets uni-axially. Sincethesematerialshave shown someextraordinaryproperties(a largeYoung's modulus, high electrical conductivity, etc.),researchers have widely usedthem invariousfields such asNanoelectronics, metalindustries, medicine and pharmacology, etc. Sincenanotubeswithdifferentdiameters, have shown different properties, controlling this factorduring thesynthesizing processisofa great importance. Inthiswork wehaveusedcatalytic chemical vapor deposition (CCVD) method [1] to synthesis CNTs on magnesiumoxide as a poroussubstrate,and investigate the effect ofsynthesizing time and temperature,onCNTs diminutions.As a result we can say withincreasingsynthesistemperature, CNTsdiameterswill increase [2]. [1]M.Kumar, Y.Ando, J. Nanosci.Nanotechnol.10, 3739 (2010). [2]L. Chen,H. Liu,K. Yang, J. Wang, X.Wang, Chem. Phys.112, 407 (2008).

NANO-22 Activation of intrinsic apoptotic signaling pathway to target breast cancer using Oxaliplatin loaded Chitosan nanoparticle S. Kannan and R.Vivek Proteomics and Molecular Cell Physiology Laboratory, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore 641 046, Tamil Nadu, INDIA

In the present study Oxaliplatin-loaded degradable chitosan nanoparticles were synthesized and characterized by TEM, DLS, Zeta Potential and the size distribution were also evaluated. Sustained release of Oxaliplatin and higher cell toxicity than free Oxaliplatin were obtained from Oxaliplatin-loaded nanoparticles. Drug release from the DDS was significantly accelerated by decreasing pH from 7.0 to 4.5, which is of particular interest to cancer therapy due to the acidic extracellular tumor environment and intracellular endosomal and/or lysosomal compartments. Drug release analysis showed a strong dependence on pH value, which was higher at lower pH. Compared with the untreated cells, Oxaliplatin loaded chitosan nanoparticles significantly upregulate the expression of Bax, Bik, Cytochrome c, caspase-9 and 3 and found to inhibit the expression of Bcl-2 and survivin in the MCF-7 cells by activating intrinsic signaling pathway induced by Oxaliplatin. Although bcl-2 and survivin are both apoptosis inhibitors, they work through different pathways in the regulation of cell apoptosis. The anti-apoptotic protein bcl-2 mainly inhibits the mitochondrial pathways, whereas the survivin directly blocks the processing and activation of caspase-3 and caspase-9, which commonly acts downstream of both apoptosis signaling pathways. Thus the present investigation suggests that Oxaliplatin loaded with chitosan nanoparticles induced apoptosis through intrinsic apoptotic pathway. Our findings clearly indicate that the chitosan nanoparticles have a strong enough potentiality to carry the loaded anticancer drug and enhanced the anticancer effect.

[1] M.K. Manion, D.M. Hockenbery, Cancer Biol Ther 2 :105 (2003) [2] A. Suzuki, T. Ito, M. Hayashida, Oncogene 19 ,1346 (2000)

NANO-23 Novel luminescent silica nanoparticles (LSN): Effect of p53 gene delivery in breast cancer in vitro and in vivo Soundarapandian Kannan Proteomics and Molecular Cell Physiology Laboratory, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore – 641 046, TN, INDIA

Mutations in the p53 tumor suppressor gene are one among the most common genetic abnormalities to be described in breast cancer. However, there are a few recant reports on non-viral vector-mediated p53 gene delivery in breast cancer. A new formulation of luminescent silica nanoparticles (LSNs) for gene delivery was produced by the two-step method with slight modification. The pp53 plasmid constructs (p53-EGFP)/LSNs complexes were transfected into human breast cancer cell (MCF-7) and transfection efficiency was determined by FACS analysis. The gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis respectively. Further the growth inhibition through induced apoptosis with pp53-EGFP/LSNs complex were assessed by trypan blue exclusion assay and annexin V staining, respectively. Interestingly the biodistribution of plasmid DNA study revealed the occurrence was investigated by PCR and RT-PCR. The transfection efficiency of LSNs showed the highest transfection efficiency among the LSN formulation was higher than that of commercially available Lipofectin®. The LSNs-mediated transfection of the p53 gene resulted in efficient high level of wild-type p53 mRNA and protein expression levels in MCF-7 cells. The efficient reestablishment of wild-type p53 function in breast cancer cells restored the p53 dependent apoptotic pathway. Taken together, our results reveal that cationic LSN-mediated p53 gene delivery may have potential application as a non-viral vector-mediated breast cancer gene therapy due to its effective induction of apoptosis and tumor growth inhibition. [1] G.Yao, L.Wang, Y.Wu, J.Smith, E.Lee, W.Tan Anal. Chem., 385, 518 (2006) [2] F. Kamangar, GM. Dores, WF.Anderson, J Clin Oncol 24, 2137 (2006).

NANO-24

Carbon Nanomaterials-based Single-Molecule Electrical Biosensors Xuefeng Guo College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China.

A universal lithographic methodology for creating functional single-molecule devices based on carbon nanomaterials as point contacts has been developed.[1] In this talk, I will detail our rational bioassay techniques by using bridge molecules with functional side groups capable of subsequent biocompatible assembly. We have been able to form complex multicomponent nanostructures from single-molecule devices by combining programmed chemical reactivity and directed self-assembly. We bridge nanogapped electrodes with a molecule that can react with a biochemical probe molecule. The probe then binds to a complementary molecule to form a noncovalent assembly. We electrically monitor each step of the process at the single event level. We have tested this approach in biological systems, including DNA hybridization, DNA-protein interaction, and biotin/streptavidin binding.[2] One key advantage of this approach to biosensing is the ability to form a well-defined chemical linkage between a molecular wire and a probe molecule. Furthermore, because it is constructed from a single molecule, each device can monitor individual binding events. This methodology demonstrates a connection

between electrical conduction and biology that offers a glimpse into the future of integrated multifunctional sensors and devices. [1] X. Guo, J. P. Small, J. E. Klare, Y. Wang, I. Tam, M. S. Purewal, B. H. Hong, R. Caldwell, L. Huang, S. O’Brien, J. Yan, R.Breslow, S. J. Wind, J. Hone, P. Kim, C. Nuckolls, Science, 311, 356 (2006); A. Feldmen, M. L. Steigerwald, X. Guo, C. Nuckolls, Acc. Chem. Res., 41, 1731 (2008). [2] X. Guo, A. Whalley, J. E. Klare, L. Huang, S. O’Brien, C. Nuckolls, Nano Lett., 7, 1119 (2007); X. Guo, A. Gorodetsky, J. K. Barton, J. Hone, C. Nuckolls, Nature Nanotech., 3, 163 (2008); S. Liu, G. H. Clever, Y. Takezawa, M. Kaneko, K. Tanaka, X. Guo, M. Shionoya, Angew. Chem. Int. Ed. 50, 8886 (2011); S. Liu, X. Zhang, W. Luo, Z. Wang, X. Guo, M. L. Steigerwald, X. Fang, Angew. Chem. Int. Ed. 50, 2496 (2011).

NANO-25 Zinc Layered Hydroxide-Salicylate Nanohybrid and Its Effect on Vero3 Cells Viability Munirah Ramli1,2, Mohd Zobir Hussein1,3,* Khatijah Mohd Yusoff4

1Advanced Materials and Nanotechnology Laboratory, Institute of Advanced Technology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia 2Department of Industrial Biotechnology, Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, Malaysia 3Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, Malaysia 4Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Malaysia

A new layered organic-inorganic nanohybrid based on zinc layered hydroxide (ZLH) which contains an anti-inflammatory agent, salicylate (SA) was synthesized through direct reaction of salicylic acid at various concentrations with commercially available zinc oxide (ZnO). Power x-ray diffraction (PXRD) patterns revealed that the basal spacing of the pure phase nanohybrid was 15.73 Å. The SA anions were arranged in a vertical bilayer form, with the carboxylate functional groups pointing towards the inorganic layers of ZLH. Fourier transform infrared (FTIR) spectrum for the synthesized nanohybrid illustrate bands characteristic of salicylate and ZLH, thereby further confirming its intercalation into the interlayers of ZLH. The loading of SA in the nanohybrid was estimated to be around 29.7%, and the nanohybrid exhibited the properties of a mesoporous-type material. Thermogravimetry and differential thermogravimetry analyses showed that the thermal stability of the SA in the nanohybrid was greatly enhanced compared to its counterpart. In vitro cytotoxicity assay revealed that free SA, pure ZnO and the nanohybrid have mild effect on viability of African green monkey kidney (Vero3) cells. NANO-26 NOSE TO BRAIN TARGETING OF OLANZAPINE CONTAINING MICROEMULSION Rashmin B. Patel*1, Mrunali R. Patel2, Kashyap K. Bhatt2, Bharat G. Patel3, Anil K Mishra3, Krishan Chutani3, Rajiv V. Gaikwad4, Abdul Samad4

1. A. R. College of Pharmacy and G. H. Patel Institute of Pharmacy, Sardar Patel University, Vallabh Vidyanagar, India. 2. Indukaka Ipcowala College of Pharmacy, Sardar Patel University, New Vallabh Vidyanagar, 388121, India. 3. CHARUSAT, Changa, Gujarat, India. 4. Ministry of Defence, Institute of Nuclear Medicine and Allied Sciences, Government of India, Timarpur, Delhi, India. 5. Veterinary Nuclear Medicine Center, Bombay Veterinary College, Parel, Mumbai, 400 012, India.

To formulate and evaluate microemulsion based drug delivery system designed for intranasal administration of Olanzapine. Drug loaded microemulsions were successfully prepared by water titration method [1, 2] and characterized for physicochemical parameters. Olanzapine microemulsions (OME) were radiolabeled with 99mTc (technetium) and biodistribution [3] of drug in the brain was studied in Swiss albino rats. Brain scintigraphy

imaging was also performed in rabbits. OME were transparent and stable with mean globule size of 20-25 nm

and zeta potential of 30mV to 40mV. 99mTc-labeled Olanzapine solution (99mTc OS)/OME/Olanzapine mucoadhesive microemulsion (OMME) were found to be stable and suitable for in vivo studies. Brain/blood uptake ratios at 0.50 hour (h) following intranasal OMME, OME, OS, and intravenous OME administrations were found to be 1.42, 1.23, 0.59, and 0.23, respectively indicating more effective targeting with intranasal administration and best targeting of the brain with intranasal OMME. Brain/blood ratio at all sampling points up to 8 h following intranasal administration of OMME compared to intravenous was found to be four fold higher indicating larger extent of distribution of the drug in brain. Drug targeting efficiency, and direct drug transport were found to be highest for OMME post-intranasal administration compared to intravenous OME. Rabbit brain scintigraphy also showed higher intranasal uptake of the drug into the brain. This investigation demonstrates a more rapid and larger extent of transport of Olanzapine into the rat brain with intranasal OMME, which may prove useful in acute and maintenance treatment of schizophrenia. [1] M.R. Patel, R.B. Patel, J.R. Parikh, A.B. Solanki and B.G. Patel, AAPS PharmSciTech 10, 917-923, (2009) [2] L. Li, I. Nandi and K. H. Kim Int. J. Pharm. 237, 77-85, (2002) [3] V.V. Jogani, P.J. Shah, P. Mishra, A.K. Mishra and A. Misra, Alzh. Dis. Assoc. Disord. 22, 116-124, (2008)

NANO-27 TRANASAL DELIVERY OF CARBAMAZAPINE USING MUCOADHESIVE MICROEMULSIONS: PRELIMINARY BRAIN TARGETING STUDIES Rashmin B. Patel1, Mrunali R. Patel2, Kashyap K. Bhatt2, Bharat G. Patel3, Anil K Mishra4, Krishan Chutani4, Rajiv V. Gaikwad5, Abdul Samad5

6. A. R. College of Pharmacy and G. H. Patel Institute of Pharmacy, Sardar Patel University, Vallabh Vidyanagar, India. 7. Indukaka Ipcowala College of Pharmacy, Sardar Patel University, New Vallabh Vidyanagar, 388121, India. 8. CHARUSAT, Changa, Gujarat, India. 9. Ministry of Defence, Institute of Nuclear Medicine and Allied Sciences, Government of India, Timarpur, Delhi, India. 10. Veterinary Nuclear Medicine Center, Bombay Veterinary College, Parel, Mumbai, 400 012, India.

To prepare and evaluate carbamazepine microemulsions (CME) for rapid drug delivery to the brain for acute and maintenance treatment of epileptic patients. The CME were prepared by the titration method [1, 2]. CME was radiolabeled with 99mTc (technetium) [3] and biodistribution of drug in the brain was studied in Swiss albino rats after intranasal and intravenous administrations. Brain scintigraphy imaging was performed in rabbits. CME were transparent and stable with mean globule size of 35-40 nm. 99mTc-labeled carbamazepine solution (99mTc CS)/CME/carbamazepine mucoadhesive microemulsion (CMME) were found to be stable and suitable for in vivo studies. Brain/blood uptake ratios at 0.50 hour (h) following intranasal CMME, CME, CS, and intravenous CME administrations were found to be 1.09, 0.66, 0.38, and 0.12, respectively indicating more effective targeting with intranasal administration and best targeting of the brain with intranasal CMME. Brain/blood ratio at all sampling points up to 8 h following intranasal administration of CMME compared to intravenous was found to be twofold higher indicating larger extent of distribution of the drug in brain. Drug targeting efficiency, and direct drug transport were found to be highest for CMME post-intranasal administration compared to intravenous CME. Rabbit brain scintigraphy also showed higher intranasal uptake of the drug into the brain. This investigation demonstrates a more rapid and larger extent of transport of carbamazepine into the rat brain with intranasal CMME, which may prove useful for acute and maintenance treatment of epileptic patients. [1] M.R. Patel, R.B. Patel, J.R. Parikh, A.B. Solanki and B.G. Patel, AAPS PharmSciTech 10, 917-923, (2009) [2] L. Li, I. Nandi and K. H. Kim Int. J. Pharm. 237, 77-85, (2002) [3] V.V. Jogani, P.J. Shah, P. Mishra, A.K. Mishra and A. Misra, Alzh. Dis. Assoc. Disord. 22, 116-124, (2008)

NANO-28 SERS-based DNA detection using inverted multi-angular Au nano-cavity array-embedded microfluidic divice C. K. Yao1, J. D. Liao1, Y. S. Yang2, S. H. Yu1, J. W. Yang1

1National Cheng Kung University, No. 1, University Road, Tainan, Taiwan 2 National Chiao Tung University, No. 1001, University Road, Hsinchu, Taiwan

Periodic nano-structured cavities arrayed on Au can induce localized surface plasmons (LSPs), which is increasingly important for the application of surface-enhanced Raman scattering (SERS) [1]. In this study, nano-indentation technique is applied to fabricate particularly made nano-structures embedded in microfluidic channel. The as-integrated device provides a convenient and reproducible platform for DNA detection. In this work, parameters for the fabrication of nano-structured surface by controlling infinitesimal indentation force and tip-to-tip displacements of each indentation step were optimized. These nano-cavities array competent to induce the LSPs effect was chosen by properly controlling the size of cavity structure that led to significantly enhance Raman signals. A specific target DNA sequence of Avian Influenza Virus (AIV) was thereafter experimented using the optimized cavity structure. At first, the complementary DNA sequence (H7-P) of target sequence (H7-T) was modified on cavity structure with thiol-modification. Qualitative analysis was examined by measuring SERS peak areas in the respective range of 700~750 cm-1 and 1100~1350 cm-1. The significant difference between H7-P hybridized with H7-T and its control could be found, with a distinguishable concentration for DNA sequence of 10-7~10-11 M. Consequently, Raman signals for target DNA sequence could be quickly analyzed using a nano-cavity array embedded in microfluidic channel for probing DNA-modified sequence. This SERS-based opto-fluidic device is thus promising for fast-screening virus. [1] C. W. Chang, J. D. Liao, A. L. Shiau, and C. K. Yao, Sensors and Actuators: B., 156/1, 471 (2011).

NANO-29 Co-encapsulation of biodegradable nanoparticles with silicon quantum dots and quercetin for monitored delivery Q. Wang1, Y. Bao2, J.H. Ahire1, Y. Chao1

1School of Chemistry, 2Norwich Medical School, University of East Anglia, Norwich, NR4 7TJ, UK. Polymer nanoparticles have emerged as a promising new strategy for the efficient delivery of drugs. They have several advantages when used as drug carriers, such as high stability, high capacity, improvement of drug bioavailability, as well as allowing for sustained drug release [1, 2]. Quercetin has therapeutic potential as an anticancer drug, but has poor solubility and low bioavailability. In this study we show that co-encapsulation of quercetin and fluorescent Silicon quantum dots (SiQDs) in Poly (ethylene glycol)-block-polylactide (PEG-PLA) nanoparticles can be used for simultaneous in vitro imaging and to improve the biocompatibility of quercetin. Fluorescent imaging with SiQDs can provide a new concept to monitor the delivery of anti-cancer drugs. The nanoparticles were synthesized based on double emulsion method and were extensively characterized and assayed for cytotoxicity in vitro. HepG2 cells were incubated with quercetin and SiQDs dual-loaded PEG-PLA nanoparticles, resulting in a red fluorescent staining which can be detected with a confocal microscope. PEG-PLA nanoparticle encapsulated quercetin suppressed human hepatoma HepG2 cell proliferation more effectively than the free standing form. In addition, nanoparticle-encapsulated quercetin significantly inhibited hydrogen peroxide-induced DNA damage in HepG2 cells. These data show that nanocapsulated quercetin possesses the potential bioactivity to reduce the drug dosage frequency, as well as increase patient compliance. The combination of polymeric nanoparticles and semiconductor quantum dots can allow monitoring of delivery,

improve aqueous solubility, and enhance biocompatibility. Such nanoparticulated systems could shape the future of drug delivery. [1] S. Sengupta, D. Eavarone, I. Capila, G. L. Zhao, N. Watson, T. Kiziltepe, R. Sasisekharan, Nature, 436, 568 (2005). [2] H. Xu, L. Cheng, C. Wang, X. Ma, Y. Li, Z. Liu, Biomater., 32, 9364 (2011).

NANO-30

Silver and DLC coated surgical mesh by magnetron sputtering technique and their bactericidal potential G.E.Testoni 1, A.M.A.Liberatore 2, P.L.Longo 3, M.Massi 1,4, A.S.da Silva Sobrinho 1, J.E.Matieli 1, C. Otani 1, I.H.J.Koh 2, L.G.Freitas Filho 2, H.S.Maciel 1,5

1 Instituto Tecnológico de Aeronáutica - ITA, 12228-900, São José dos Campos, Brasil 2Universidade Federal de São Paulo, Campus São Paulo, 04021-041, São Paulo, Brasil 3 Universidade de São Paulo, ICB II, 05508-900, São Paulo, Brasil 4Universidade Federal de São Paulo-ICT, Campus SJC , 12.231-280, São José dos Campos, Brasil 5Universidade do Vale do Paraiba, 12.244.000 - São José dos Campos

The use of meshes for treatment of the large abdominal damage has increased significantly since Usher et al first used the polypropylene mesh in 1958. Despite the continued spread of its use, the available meshes cannot be considered ideal in face of possible organic reactions of the host to material antigenic, which can cause infection up to 8% of cases and other complications such as intestinal fistulae and obstruction, resulting in serious physical and psychosocial consequences. In this study, the surgical polypropylene meshes (PM) were coated with diamond like carbon (DLC) or with silver (Ag) thin films by DC magnetron sputtering aiming to confer biocompatibility and bactericidal properties, respectively. Initially we evaluated the DLC and Ag bactericidal effect. The DLC and Ag coated PM were sterilized by gamma radiation (15 kGy) and tested for bacterial sensitivity using the adapted disk diffusion method in Mueller-Hinton agar plate inoculated with E. coli, and incubated for 24 h at 37°C. The silver coated mesh promoted a clear bacterial inhibition zone and suggestive bactericidal effect between the braids of the mesh, while in carbon coated mesh the bacterial growth occurred uniformly in the entire mesh area, showing the biocompatibility of the carbon to bacteria. New assays with silver doped DLC thin films are being processed to find the minimum bacterial inhibition layer. Based on these preliminary findings the magnetron sputtering deposition technique has the potential to bring new contributions in the development/improvement of the medical devices. Other experiments are being processed to evaluate the biocompatible and bactericidal combined effect of the surgical mesh coated with carbon and silver. NANO-31 Cadmium Free Quantum Dots M. Booth1 and K. Critchley1

1School of Physics and Astronomy, University of Leeds, Leeds, LS2 9JT, United Kingdom

Quantum dots show great promise for bio-imaging and a great deal of progress has been made. However, a vast majority of studies are performed using CdX or PbX based nanoparticles and this raises fears of their potential toxicological effects, both in manufacture (environmental), handling and administering the nanoparticles in-vivo. We have been developing alternative quantum dots that do not contain Cd or Pb, have an ideal emission range, have a similar photoluminescent quantum yield to Cd-based quantum dots, whilst also being able to adopt similar surface chemistry protocols as used for CdSe quantum dots. The nanoparticles we have choosen to study

(a) (b)

are copper indium disulphide1. We show that we can synthesize CuInS2/ZnS quantum dots, modify their surface chemistry by synthesizing amphiphilic polymers2 to enable them to be disperse in the aqueous environment, and finally we have started to in vitro study their uptake and toxicity. [1] M. Booth, AP Brown, SD Evans, K Critchley Chem. Mater. 2012, 24, 2064−2070 [2] C-AJ Lin et al Small,4,3,334-341.

NANO-32 Multifunctional Nanoplatform for Biomedical Applications R. Piñol1, L. Mohamed1, R. Bustamante1, C. Brites2, L. Gabilondo1, J.L. Murillo1, N.J.O. Silva2, V. Sorribas3, M. Gutierrez3, R. Cornudella3, L. Carlos2, F. Palacio1, A. Lascialfiari4, J. Carrey5, M. Respaud5, G. C. Sanmartí6, J.-P. Salvador6, P. Marco6, J. Criado7, M. Fuentes7, A. Millan1

1 ICMA, Zaragoza, C/ Pedro Cerbuna 10, 50009 Zaragoza, Spain; 2 University of Aveiro, 3810–193Aveiro, Portugal; 3 University of Zaragoza, C/ Pedro Cerbuna 10, 50009 Zaragoza, Spain 4 University of Pavia, Pavia, Italy; 5 INSA, 135, av. de Rangueil, F-31077 Toulouse, France; 6 IQAC-CSIC, Jordi Girona, 18-26, 08034, Barcelona, Spain; 7 CIC, Campus Miguel de Unamuno, 37007 Salamanca. España,

Working machinery in life is nanometric, thus, it is no wonder that the development of adequate nanotools would be very helpful in biomedical science. In this direction, the idea behind this work is to build a nanoplatform that can incorporate, in an easy way, multiple physical and biological functionalities. The core of the platform is an hydrophobic polymer that may be used as a matrix for the encapsulation of inorganic nanoparticles (magnetic, luminescent, radioactive, …). This matrix contains a Michael-donor (or an acceptor) on its surface for functionalization. Organic bioactive molecules are attached to one end of a hydrophilic polymer (i.e. PEG) terminated on a Michael acceptor (or a donor), and then they are anchored to the hydrophobic core by Michael addition. This system has the advantages of a clean synthesis (no by-products), mild conditions, and an easy and controlled multifunctionalization. So far, we have incorporated to this platform: magnetic nanoparticles, optical dyes (fluorescein and rhodamine), a therapeutical drug, an antibody, and an optical thermometer made of lanthanide complexes. Health safety of the system has been tested in cellular and in vivo assays. The nanoplatform is highly stable in biological fluids, shows low cell toxicity, high capacity of cell internalization, excellent hematocompatibility, and anticoagulation properties. It is shown that magnetic properties can be tuned up in the whole superparamagnetic range. Moreover, the system has shown excellent performance in magnetic resonance imaging and hyperthermia. NANO-33 Ferromagnetic Carbon Nanotubes for Cancer Treatment G. Raniszewski, L. Szymanski, Z. Kolacinski Technical University of Lodz, Stefanowskiego street 18/22, 90-924 Lodz, Poland

In some cases surgery of a cancer covering a large area is impossible or too risky. One of the least invasive method for cancer cells treatment employ a radio frequency (RF) wave. RF ablation (RFA) is also used e.g. in the treatment of liver cancer and cancer metastasis from other organs such as breast, colon and lung. The method uses an electrical current of high frequency to heat and destroy cancer cells. Nanoparticles with ferromagnetic

element are attached to the abnormal cells. A metal particles placed on surface or inside the nanotubes begin to heat the cancer cell under radio waves with a suitable frequencies nanotubes. After reaching a certain temperature, a cancer cell is destroyed without affecting normal cells [1-2]. RF therapy is characterized by a lower risk than other techniques for the removal of tumor cells. This method may also become the main method of treatment of small tumors and can provide a faster and more targeted cancer treatment with fewer side effects compared with conventional methods [3]. Furthermore, if carbon nanotubes will have magnetic properties, it would facilitate the manipulation of them within the body. Combination of experiences from multidisciplinary areas – material science, biotechnology, medicine, chemistry – will enable development of new treatment methods. Acknowledgements Scientific work has been financed from the budget for science in the years 2010-12 as a research project. [1] C.J. Gannon, P. Cherukuri, B.I.Yakobson, L.Cognet, J.S.Kanzius, C.Kittrell, R.B.Weisman, M.Pasquali, H.K.Schmidt, R.E.Smalley,

S.A.Curley, Cancer. 2007 Dec 15;110(12):2654-65. [2] United States Patent No.: US 7,510,555 B2 Enhanced systems and methods for RF-induced hyperthermia [3] H.C.Fernando, A.D.Hoyos, R.J.Landreneau, S.Gilbert, W.E.Gooding, P.O.Buenaventura, N.A.Christie, C.Belani, J.D.Luketich, The Journal of Thoracic and Cardiovascular Surgery, Vol. 129, Issue 3 , 639-644

NANO-34 Carbon nanotubes and electrical devices for cancer treatment and therapy G. Raniszewski Technical University of Lodz, Stefanowskiego 18/22 str., 90-924 Lodz, Poland

One of the main areas for nanotechnology achievements is medicine. One of the greatest challenges of the twenty-first century medicine is the development of treatment options for cancer. The aim of this work is to present the synthesis of carbon nanotubes that could be used as elements supporting the fight against cancer. Carbon nanotubes (CNTs) have unique properties such as mechanical strength, high thermal conductivity, high electrical conductivity. The small size of the nanotubes allows for application in drug delivery systems. They can penetrate cell membranes and penetrate into the cytoplasm or the nucleus. The literature describes several examples of delivery of therapeutic agents [1]. For example, one of the chemotherapeutic drugs are amphotericin (AMB). SWNTs can also be used in boron-neutron capture therapy (BNCT) for treatment of some types of cancer, especially brain. Another application of CNTs in cancer therapy is its ability to absorbtion of radiation. Cancer cells attached to carbon nanotubes with ferromagnetic particle - such as iron - treated with a suitable radio frequency heat up the nanotubes, and in final effect overheat cancer cells leading to their destruction. So far, decomposition, toxicity and influence on tissues are still under investigation. This paper reviews the possibility of using carbon nanotubes in the cancer diagnosis and treatment. Electrical devices for treatment by hyperthermia are described. The requirements for nanotubes for medical applications are presented. Methods of carbon nanotubes synthesis are compared. Acknowledgements Scientific work has been financed from the budget for science in the years 2010-12 as a research project. [1] C. Kumar, Ed, Nanomaterials for the Life Sciences, Vol. 6, (Wiley-VCH, Weinheim, 2006)

NANO-35 Supramolecular assemblies of cationic lipid and gramicidin as microbicidal agents D. A. M. T. Ragioto1, A. M. Carmona-Ribeiro1

1Universidade de São Paulo, São Paulo, Caixa Postal 26077, 05513-970, São Paulo, Brazil,

Dioctadecyldimethylammonium bromide (DODAB) is an antimicrobial lipid that forms large bilayer vesicles (LV) or bilayer fragments (BF) depending on the dispersion method1,2. Gramicidin (Gr) is a model antimicrobial peptide which increases the bilayer permeability towards cations. Here combinations of DODAB bilayers and Gr are shown to be very effective against Staphylococcus aureus and Escherichia coli. The assemblies were characterized by filtration for determining Gr incorporation in the bilayers, spectroscopic methods, dynamic light-scattering for sizing and zeta-potential analysis. DODAB/Gr antimicrobial activity was evaluated by plating and CFU counting. There is extensive incorporation of Gr in the cationic bilayers and occurrence of functional channels in LV/Gr assemblies. Gr incorporated in BF senses a more polar environment than in LV. Gr increases the bilayer charge density in LV but does not affect the BF charge density, showing Gr localization at the BF borders. The dissipation of osmotic gradients across the LV/Gr bilayer shows that the peptide is in the channel conformation. The resistance of S. aureus towards quaternary ammonium amphiphiles was reported3 and also observed for DODAB against S. aureus. However, for the DODAB/Gr combination, there is a tenfold reduction in the DODAB doses required to kill S. aureus (at 0.23 -0.32 μM of Gr). On the contrary, E. coli remains very sensitive to DODAB but is barely killed by Gr. The combination reduced the effective Gr doses for 99% killing by about 20 times (at 6-8 μM of DODAB). DODAB dose in BF/Gr for 99% killing is always smaller than the one in LV/Gr suggesting the superior ability of BF to deliver Gr. [1] A.M. Carmona-Ribeiro, Chem. Soc. Rev., 21, 209 (1992). [2] A.M. Carmona-Ribeiro, Int. J. Nanomedicine, 5, 249 (2010). [3] M. Otto, Nat. Rev. Microbiol., 7(8), 555 (2009).

NANO-47 Tadalafil Nanocrystals: A Novel Nanofabrication Approach for Solubility and Dissolution Rate Enhancement M.M. Mehanna, A.N. Allam Faculty of Pharmacy, Alexandria University, Alexandria, Egypt

Tadalafil is used to treat erectile dysfunction. Tadalafil is characterized by low solubility and high permeability (a class II drug), which has a negative influence on its bioavailability. Utilization of nanocrystals formation approach represents an effective and facile methodology to enhance the poorly soluble drugs dissolution rates. Our aim was to select and optimize various parameters that affect the preparation of tadalafil nanocrystals, which were prepared by the antisolvent precipitation method. Preliminary studies were carried using Pluronic F-127, Pluronic F-68, Hydroxypropyl methylcellulose (50 and 4000 c.p.), polyvinyl chloride, and polyvinyl alcohol as stabilizers. In order to shed more lights on the optimum conditions leading to the preparation of tadalafil nanocrystals, the effects of the studied parameters on crystal size, solubility and dissolution rate were investigated. The results obtained pointed out that the use of methanolic solution of tadalafil to water as the antisolvent produced crystals in the submicron range in presence of a stabilizer. The particle size decreased with the increase of the stabilizer concentration but further increase of which didn't affect the crystals size significantly. IR spectroscopical studies revealed that there was no chemical changes in tadalafil structure occurred during nanofabrication. The differential scanning calorimetry (DSC) analysis illustrated that there was no substantial

crystalline change in the nanocrystals compared to the raw drug. Both the solubility and the in-vitro dissolution rate of tadalafil were significantly augmented through introduction of the nanocrystals. NANO-48 Nanocapsules Based Pediatric Liquid Lisinopril Formulation: Design and Physicochemical Characterization M. M. Mehanna Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.

Lisinopril is an orally active angiotensin-converting enzyme (ACE) inhibitor used for the treatment of hypertension, heart failure, and acute myocardial infarction in infants. It is currently supplied as tablets. Solid dosage forms represent a problem in children with swallow difficulty and in neonates. There is no commercially available oral liquid preparation of lisinopril. Our strategy is to prepare oral liquid formulation of lisinopril based on lipid-core nanocapsules. Nanocapsules were prepared according to the nanoprecipitation protocol utilizing poly-ε-caprolactone as a shell-forming polymer. The following characteristics of nanocapsules were determined: encapsulation efficiency, nanocapsules size, in-vitro drug release and stability on storage. The results revealed that the optimized formulations of lisinopril-loaded nanocapsules were in the nanometric range with a high encapsulation efficiency (more than 90%), both stable for 3 months. The release of lisinopril from the polymer-based nanocapsules was rapid in the simulated gastric fluid. In the top of that, the concentration of lisinopril in the liquid preparation was appropriate for oral pediatric administration. NANO-49 Iron-filled carbon nanotubes for magnetic hyperthermia cancer therapy M. Baxendale School of Physics and Astronomy, Queen Mary, University of London, Mile End Road, London E1 4NS, UK.

Carbon nanotubes have many potential applications in drug delivery systems and regenerative medicine. Magnetic nanoparticles for magnetic hyperthermia cancer therapy must operate under structural, chemical, thermal, and operational constraints. Iron-filled carbon nanotubes can satisfy the requirements with some careful control of the phase of the iron content and development of functionality. This report outlines the development of an iron-filled carbon nanotube-based magnetic hyperthermia system optimized for magnetic response and chemical integrity with routes to dimensional control, integrated thermometry, and MRI contrast agent. NANO-50 A new material for orthopaedics application with surface nanotopography Farzad Shabani, Alexander M. seifalian University College London, Centre for Nanotechnology & Regenerative Medicine, UCL Division of Surgery & Interventional Science, Royal Free Hampstead NHS Trust Hospital, United Kingdom We have developed and patented a family of nanocomposite materials for surgical implants. The materials based on polyhedral oligomeric silsesquioxane (POSS) nanoparticle and poly(carbonate urea)urethane (PCU).

This polymer has exhibited enhanced biocompatibility, biostability, and anti-thrombogenic tested in vitro. Here we present the in vivo biocompatibility of POSs-PCU. These polymers were implanted in six healthy sheep (n = 6) for 36 months and a siloxane served as the positive control. After explantation, these polymers were extracted, as was the surrounding capsule, if any. Attenuated total reflectance Fourier transform infrared spectroscopy analysis was performed to look for signs of surface degradation on the polymers and histopathologic and electron microscopic examinations were performed to study the interaction between the biomaterial and the host environment in greater detail. After implantation, the authors observed minimal inflammation of the nanocomposite within the sheep model as compared with the siloxane control. Contact angle measurements and fibrinogen enzyme-linked immunosorbent assay tests were then conducted on the POSS-PCU nanocomposite to determine the reason for this behaviour. The increased fibrinogen adsorption on POSS-PCU, its amphilicity, and large contact-angle hysteresis indicated that POSS-PCU inhibits inflammation by adsorbing and inactivating fibrinogen on its surface. In complete contrast, the control siloxane in the same setting demonstrated very significant inflammation and degradation, resulting in capsular formation. Naturally, there was no evidence of degradation of the nanocomposite compared with the siloxane control. POSS-PCU nanocomposites have enhanced interfacial biocompatibility and better biological stability as compared with conventional silicone biomaterials, thus making them safer as tissue implants. NANO-51 Inception to Actualization: Next Generation Coronary Stent Coatings incorporating Nanotechnology A. Tan1*, Y. Rafiei1, A. De Mel1, J. Rajadas2, M.S. Alavijeh, A.M. Seifalian1,3 1Centre for Nanotechnology & Regenerative Medicine, UCL Division of Surgery & Interventional Science, University College London, London, UK 2Biomaterials & Advanced Drug Delivery Laboratory, Department of Chemical Engineering, Stanford University, USA 3Pharmidex Pharmaceutical Services Ltd., London, UK 4Royal Free London NHS Foundation Trust, London, UK

Coronary artery disease is a leading cause of death globally. Percutaneous coronary intervention (PCI) utilizing balloon angioplasty and coronary stents are used to treat blocked coronary arteries. Early versions of stents were classified as bare-metal stents (BMS) and faced issues of in-stent restenosis, which resulted in a re-narrowing of the stented vessel, necessitating repair procedures. Drug-eluting stents (DES) were developed in response to the issues seen in BMS, as anti-proliferative drugs inhibited vascular smooth muscle cell growth, thereby preventing restenosis. However the use of DES engendered another problem: late stent thrombosis (ST), which is a potentially fatal complication. Studies have shown that late ST is partly attributed to inflammation due to drug/polymer matrix hypersensitivity. We have previously developed and patented a novel nanocomposite polymer, polyhedral oligomeric silsesquioxane poly (carbonate-urea) urethane (POSS-PCU), which is highly biocompatible and currently used in patients as artificial trachea, lacrimal duct and bypass graft. We therefore postulate that coating stents with POSS-PCU via an optimized ultrasonic atomization spray technique could potentially circumvent current problems seen in BMS and DES. Incorporating nanotechnology, we seek to explore the attachment of endothelial progenitor cell (EPC)-specific antibodies onto POSS-PCU for in situ endothelializataion; polymer biofunctionalization using nitric oxide (NO) donors; and drug/polymer multilayers for controlled drug release. The convergence of EPC capture technology, polymer biofunctionalization and controlled drug release would form the cornerstone of what we advocate is the next generation nanocomposite polymer coatings for coronary stents.

NANO-52 Potential of electrospun polyurethane blended with elastin and collagen as vascular graft C.S. Wong, X. Liu, Z. Xu, T. Lin and X. Wang Institute for Frontier Materials, Deakin University, Geelong, Victoria, 3217, Australia.

The mismatch in mechanical properties between synthetic vascular graft and native arteries is one of the factors that contribute to graft failure. Polyurethane is a suitable choice of material for vascular grafts as it has good haemocompatibility and elasticity. However, its mechanical properties are still not comparable to native arteries. The viscoelastic properties of native arteries are conferred by elastin and collagen. In this present study, aligned nanofibrous scaffolds consisting of polyurethane blended with elastin or a mixture of elastin and collagen type I were created via electrospinning. The cellular interactions and mechanical properties of these protein blended polyurethane scaffolds were examined. The unmodified polyurethane scaffold exhibited a peak stress of 33.44 MPa and a strain of 246.68% indicating a material of high strength and flexibility. The addition of 5% elastin to polyurethane reduced the peak stress and strain to 7.86 MPa and112.28% respectively; characteristics that are similar to those observed in blood vessels. Blending of both elastin and collagen to polyurethane strengthened the scaffold and increased its peak stress to 28.14 MPa when compared to elastin blended scaffolds. Human coronary artery smooth muscle cells were cultured on various types of scaffold for 7 days and cell growth increased on the elastin and collagen blended scaffold (8.70 x 103 ± 2.86 x 103 cells). Immunostaining of the cell-seeded scaffolds with antibody against smooth muscle myosin demonstrated that the smooth muscle cells displayed a contractile phenotype which is preferred for tissue engineering applications. The addition of elastin and collagen to polyurethane is beneficial towards the creation of compliant synthetic vascular grafts as elastin provided the necessary viscoelastic properties while collagen imparted the biological cues required for enhanced cellular interaction. NANO-53 Application of nano composite material IN orthopaedics Farzad Shabani, Alexander M. seifalian University College London, Centre for Nanotechnology & Regenerative Medicine, UCL Division of Surgery & Interventional Science, Royal Free Hampstead NHS Trust Hospital, United Kingdom

We have developed and patented a family of nanocomposite materials for surgical implants. The materials based on polyhedral oligomeric silsesquioxane (POSS) nanoparticle and poly(carbonate urea)urethane (PCU). This polymer has exhibited enhanced biocompatibility, biostability, and anti-thrombogenic tested in vitro. Here we present the in vivo biocompatibility of POSs-PCU. These polymers were implanted in six healthy sheep (n = 6) for 36 months and a siloxane served as the positive control. After explantation, these polymers were extracted, as was the surrounding capsule, if any. Attenuated total reflectance Fourier transform infrared spectroscopy analysis was performed to look for signs of surface degradation on the polymers and histopathologic and electron microscopic examinations were performed to study the interaction between the biomaterial and the host environment in greater detail. After implantation, the authors observed minimal inflammation of the nanocomposite within the sheep model as compared with the siloxane control. Contact angle measurements and fibrinogen enzyme-linked immunosorbent assay tests were then conducted on the POSS-PCU nanocomposite to determine the reason for this behaviour. The increased fibrinogen adsorption on POSS-PCU, its amphilicity, and large contact-angle hysteresis indicated that POSS-PCU inhibits inflammation by adsorbing and inactivating fibrinogen on its surface. In complete contrast, the control siloxane in the same setting demonstrated very significant inflammation and degradation, resulting in capsular formation. Naturally,

there was no evidence of degradation of the nanocomposite compared with the siloxane control. POSS-PCU nanocomposites have enhanced interfacial biocompatibility and better biological stability as compared with conventional silicone biomaterials, thus making them safer as tissue implants. NANO-54 Hybrid Ceramo-Polymeric Modified Dental Implants with Improved Biomimetic Characteristics A. Apicella, R. Aversa Second University of Naples, Aversa, Abbazia di San Lorenzo 81031, ITALY.

The research develops and tests new hybrid biomimetic materials that work as mechanically stimulating "scaffolds" to promote early regeneration in implanted bone healing phases. A biomometic nanostructured osteoconductive material coated apparatus is presented. Bioinspired approaches to materials and templated growth of hybrid networks using self-assembled hybrid organic-inorganic interfaces is finalized to extend the use of hybrids in the medical field. Combined in vivo, in vitro and computer-aided simulations have been carried out. A new experimental methodology for the identification of design criteria for new innovative prosthetic implant systems is presented. The new implant design minimizes the invasiveness of treatments while improving implant functional integration [1]. A new bioactive ceramo-polymeric hybrid material was used to modify odontostomatological Titanium implants in order to promote early fixation, biomechanical stimulation for improved scaffold mineralization and ossification. It is a hybrid ceramo-polymeric nanocomposites based on Hydroxyl-Ethyl-Methacrylate polymer (pHEMA) filled with nanosilica particles that have shown biomimetic characteristics [2,3]. This material swells in presence of aqueous physiological solution leading to the achievement of two biomechanical functions: prosthesis early fixation after and bone growth stimulation. Such multidisciplinary approach explores novel ideas in modelling, design and fabrication of new nanostructured biomaterials with enhanced functionality and improved interaction with OB cells. [1] R Aversa, D Apicella, L Perillo, R Sorrentino, F Zarone, M Ferrari, A Apicella, Dental Materials, 25(5), 678-690 (2009). [2] C Schiraldi, A D'Agostino, A Oliva, … R Aversa, M De Rosa, Biomaterials, 25 (17), 3645-3653 (2004). [3] G R. Beck, Shin-Woo Ha, C E Camalier, M Yamaguchi, Y Li, J K Lee, M. N Weitzmann, Nanomedicine, 8(6), 793-803 (2012).

NANO-55 Measuring the viscous and elastic properties of single cells using particle tracking microrheology R.L. Warren, M. Tassieri, X. Li, A. Glidle, D.J. Paterson, A. Carlsson, and J.M. Cooper Division of Biomedical Engineering, University of Glasgow, Glasgow, U.K.

The mechanical properties of a cell's cytoskeleton can influence factors such as growth, apoptosis, motility, signal transduction and gene expression. Related to this, there is a desire to be able to provide a rheological interpretation of the cell's viscoelastic properties that has the potential to yield quantitative information on the cell's cytoskeletal structure and dynamics. The interpretation of rheology measurements performed on living systems such as cells is not trivial and represents a lively point of discussion in literature. The debate is centred on the violation of the fluctuation-dissipation theorem (FDT) because of the existence of non-thermal forces, generated by protein-protein interactions (e.g. motor proteins), governing the cells’ response at relatively low

frequencies (i.e. 10 Hz). However, at high frequencies (i.e. 10 Hz), all the competing theories together with the related experimental evidence agree on the fact that in the cells the thermal fluctuations dominate on those induced by protein-protein interactions; i.e. the cytosckeleton can be considered as if it were at thermal

equilibrium, thus the applicability of the FDT. In this work we present a simple and non-invasive experimental procedure to measure the in vivo linear viscoelastic properties of cells by passive particle tracking microrheology. In order to do this, a generalised Langevin equation is adopted to relate the time-dependent thermal fluctuations of a probe sensor, immobilised to the cell's exterior, to the frequency-dependent viscoelastic moduli of the cell. The method has been validated by measuring the linear viscoelastic response of a soft solid and then applied to cell physiology studies. It is shown that the viscoelastic moduli are related to the cell's cytoskeletal structure, which in this work is modulated either by inhibiting the actin/myosin-II interactions by means of blebbistatin or by varying the solution osmolarity from iso- to hypo-osmotic conditions. The insights gained from this form of rheological analysis promises to be a valuable addition to physiological studies; e.g. cell physiology during pathology and pharmacological response. NANO-56 New Microcapsules As Carriers For Effective Therapy Of Different Kinds Of Tuberculosis M.K. Gilmanov1, A.N. Begzat1, S.A.Ibragimova1, A.B.Karabalin2, L.K. Amanzholova2, S.O. Tutkishbaev2

1Scientific Research Institute of the Problems of Biology and Biotechnology of Al-Faraby’s Kazakh National University, Almaty, Al-Faraby avenue 71, Kazakhstan 2National Centre For Problems of Tuberculosis, Almaty, Bekhozhin street 5, Kazakhstan

According to the World Health Organization, more than two billion people are infected with mycobacterium tuberculosis. Success in fighting with this dangerous disease could be achieved by using methods of the nanomedicine. The most promising method of nanomedicine for therapy of diseases are nanocarriers. However nowadays existing nanocarriers have serious disadvantages, for this reason nanocarriers haven't found wide application in practical medicine. Thus, our task is the development of new nanocarrier from natural material for successful therapy of the different kinds of tuberculosis. We were developed the method of the preparation new microcapsules for therapy of tuberculosis. The obtained microcapsules were loaded by antitubercular antibiotics. Then the loaded microcapsules were mixed with ointment in equal proportion for preparation of the nanoointment. At first the experiments were carried out on guinea pigs. The suspension of the culture of Mycobacterium tuberculosis was injected into knee joint of the guinea pigs. After one month the guinea pigs were taken oss tuberculosis. The control animal were cured by tablets of antibiotics in quantity which corresponded to the weight of the experimental animal. For therapy of the experimental animals the nanoointment were rubbed on the shaved skin of the sick knee two times per day during three months. Two guinea pigs from control group were died before the end of the experiments whereas all animals of experimental group are stayed alive. It was established that therapy by nanoointment was much better than therapy by tablets. After three months all animals from experimental group were recovered from oss tuberculosis whereas the animal from control group are stayed ill. Also, we carried out the therapy of oss and lung tuberculosis by our nanoointment on volunteers patients. For example: volunteer patient with spinal tuberculosis who receives therapy by nanoointment is recovered during three months whereas traditional therapy demands 8-9 months.

NANO-57 Transcytosis of glucose-coated gold nanoparticles across human brain endothelium R. Gromnicova, H.A. Davies, P. Sreekanthreddy, F. Colyer, J.B. Phillips, D.K. Male Biomedical Research Network, The Open University, Milton Keynes, MK7 6AA.

Treatment of CNS disease is severely restricted because the blood-brain barrier prevents >95% of potentially useful drugs from entering the brain. The barrier is formed by the brain microvascular endothelium, which has continuous tight junctions and an array of ABC-transporters that actively efflux many xenobiotics. Brain endothelium also has a set of metabolite transporters, including the glucose-transporter Glut-1, which have been exploited in different ways to circumvent the barrier. The aim of this study was to investigate whether 5nm glucose-coated gold nanoparticles would cross human brain endothelium more efficiently than non-brain endothelium. The rate of transcytosis was compared in vitro, in primary human brain endothelium, the brain endothelial line hCMEC/D3, and two non-brain endothelial cell lines. Transcytosis was measured by transmission electron microscopy (TEM), counting nanoparticles localised between the basal plasma membrane of the endothelium and the basal lamina over 0-8hrs. The nanoparticles crossed the endothelium, by moving through the cytosol; they were not seen in inter-cellular junctions or in vesicles or in the nuclei of the endothelium. Nor did antibiotics that block vesicular transport pathways inhibit movement. The rate of movement of the nanoparticles across brain endothelium was 20-50x faster than across non-brain endothelium. The results indicate that that these nanoparticles selectively cross brain endothelium, by a biophysical process that involves movement across the apical and basal plasma membranes of the endothelium. To demonstrate that these nanoparticles could deliver a cargo to glial cells, we have used a newly-developed 3-dimensional cell culture system, which includes human astrocytes in a 3D gel matrix overlaid with a brain endothelial cell monolayer (hCMEC/D3). Transmission EM, showed that the nanoparticles cross the brain endothelium and migrate through the gel matrix, to become localised in the nuclei of the astrocytes. NANO-58 A Versatile and Widely Applicable Strategy for the Conjugation of Quantum Dots to Biomolecules through Click Chemistry A Bestetti1, C Schieber1, J Lim1, A. Ryan1, T Nguyen1, P A Gleeson1, P S Donnelly1, S J Williams1 and P Mulvaney1

1The University of Melbourne, Parkville 3010, Victoria, Australia

The use of Quantum Dots (QDs) as an alternative to fluorescent organic dyes for the labelling of biomolecules has attracted a significant amount of interest in recent years. Due to their unique spectral and physical properties, QDs promise to overcome some of the limitations of organic dyes, such as broad emission profiles and low photobleaching thresholds. However, in order for QDs to realise their full potential in biological labelling, a reliable conjugation strategy that addresses simultaneously the issues of water-solubility and stable linkage with the target needs to be developed. Until now, a wide number of methods have been proposed but only a few exhibit all the desirable characteristics, such as a wide applicability, a fast and stable linking mechanism, bioorthogonality and the ability to control the number of attached biomolecules. Here we present a protocol that addresses many of these issues. CdSe/ZnS core-shell QDs are rendered water-soluble and biocompatible through a polymer encapsulation technique that ensures colloidal stability over a wide range of buffer conditions, without altering the spectral properties of the nanocrystals. In addition, it allows the nanocrystals surface to be equipped with a tunable amount of various functional groups (e.g. azides) and assures low levels of non-specific binding. The conjugation to biological targets is achieved through the use of a bi-functional linker molecule that is able to react with amine groups present on biomolecules and, via click-chemistry, with the azide groups

present on the QDs. This linking strategy is quick, stable and of wide applicability. Furthermore, due to the possibility of controlling the amount of linkers present on the target and the amount of functional groups present on the QDs, it provides a high degree of flexibility adaptable to different experimental settings. Transferrin, a glycoprotein involved in cellular iron uptake, was chosen as a model to test the strategy. During the talk, each step of the protocol will be characterized, showing a high degree of control over the entire conjugation process. Even if bound to QDs, Transferrin appeared to retain its activity; conjugates were able to bind to Transferrin receptors placed on the surface of the cells and to become internalized, without apparent disruption of the usual internalization pathway. We will conclude with a discussion about possible improvements of the strategy with regard to the orientation of biomolecules on the QDs surface. [1] W R Algar, D E Prasuhn, M H Stewart, T L Jennings, J B Blanco-Canosa, P E Dawson, I L Medintz, Bioconj. Chem., 22, 825-858 (2011) [2] J van Embden and P Mulvaney, Langmuir, 21, 10226-10233 (2005) [3] E E Lees, T Nguyen, A H A Clayton, B W Muir and P Mulvaney, ACS Nano, 3, 5, 1121-1128 (2009)

NANO-59 Enzyme-cleavable hyaluronic acid nanocapsules containing polyhexanide and their exposure to bacteria G. Baier, A. Musyanovych and K. Landfester Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.

The aim of the BacterioSafe (“BacterioSafe” grant#245500) consortium is to develop a unique active wound dressing, which incorporates novel colorimetric sensor and active therapeutic processes for detecting and counteracting pathogenic bacteria in wounds. For this purpose, the development of biocompatible hyaluronic acid nanocapsules which show the required sensitivity and efficacy and at the same time considerably stability is reported. The formation of different types of polymeric nanocapsules (NCs) was achieved through a polyaddition reaction at the miniemulsion droplets interface. This technique allows the synthesis of stable nanocapsules whose properties can be tailored individually according to specific requirements [1,2,3]. The nanocapsule size (between 150 and 500 nm) and the stability of the system can be adjusted by varying the amount of the biomolecule, surfactant and cross-linker. After the synthesis the nanocapsules were transferred in an aqueous medium and investigated in terms of size, size distribution, functionality, and morphology using dynamic light scattering (DLS), zeta potential measurements and scanning electron microscopy (SEM). To investigate the enzyme triggered deletion of the different polymeric shells a fluorescent dye (sulforhodamine 101) was encapsulated and monitored using fluorescence spectroscopy. The release of polyhexanide and the cleavage of nanocapsules was detected using UV spectroscopy. Additionally, the stability of the nanocapsules in several biological media and the interaction of nanocapsules with human serum protein by isothermal titration calorimetry (ITC) were studied. Finally, the antibacterial activities of the nanocapsules were studied in the presence of Staphylococcus aureus strains ATCC 29213 and ATCC 43300, and Escherichia coli strain ATCC 25922. [1] Landfester, K., Angewandte Chemie 2009, 121, 4556-4576. [2] Baier, G.; Musyanovych, A.; Landfester, K., Biomacromolecules 2010, 11, (4), 960-968. [3] G. Baier, D. Baumann, J.-M. Siebert, A. Musyanovych, . Mail nder, K. Landfester Biomacromolecules 2012, DOI: 10.1021/bm300653v

NANO-60 Functionalized biodegradable nanocapsules and their cellular uptake in different cells G. Baier, A. Musyanovych and K. Landfester Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.

Synthesizing nanocarriers with stealth properties and delivering a “payload” to the particular organ remains a big challenge but is the prime prerequisite for any in vivo application. As a non-toxic alternative to the modification by poly(ethylene glycol) PEG, we describe the synthesis of crosslinked hydroxyethyl starch (HES, Mw 200,000 g/mol) nanocapsules with a size range of 170-300 nm which do not show non-specific uptake into cells. The specific uptake was shown by coupling a folic acid conjugate as a model targeting agent onto the surface of the nanocapsules, since folic acid has a high affinity to a variety of human carcinoma cell lines which overexpress the folate receptor on their surface. The covalent binding of the folic acid conjugate onto HES capsules was confirmed by FTIR and NMR spectroscopy. The coupling efficiency was determined using fluorescence spectroscopy [1]. In addition, a one-step synthesis of poly(butyl cyanoacrylate-co-propargyl cyanoacrylate) (P(BCA/PCA)) and polyurethane (PUR)-azide nanocapsules for direct covalent and variable functionalization was studied. Both types of nanocapsules were synthesized in inverse (water-in-oil) miniemulsion through either interfacial anionic polymerization (P(BCA/PCA)) or interfacial polyaddition reaction (PUR-azide). The presence of propargyl and azide groups on the surface of P(BCA/PCA) and PUR nanocapsules, respectively, enables to achieve high yields of functionalization using copper-free click-chemistry in order to avoid any cytotoxic and side effects related to the copper. [1] G. Baier, D. Baumann, J.-M. Siebert, A. Musyanovych, V. Mail nder, K. Landfester Biomacromolecules 2012, DOI: 10.1021/bm300653v [2] Baier, G., Siebert J. M., Landfester, K., Musyanovych, A., Macromolecules 2012, 45(8): 3419-3427.

NANO-61 Characterising the interaction between silver/gold nanoparticles and recombinant form of arginine kinase of Trypanosomabrucei (TbAK) O.S Adeyemi1,2, C.G Whiteley1

1Depeartment of Biochemistry, Microbiology & Biotechnology, Rhodes University, Grahamstown – 6140, South Africa. 2Department of Chemical Sciences, Redeemer’s University, Redemption Camp, Mowe – 121001, Nigeria. Presenting author: O.S. Adeyemi, Depeartment of Biochemistry, Microbiology & Biotechnology, Rhodes University, Grahamstown – 6140, South Africa.

Study investigated the interaction between a recombinant form of the arginine kinase and metal (Ag/Au) nanoparticles or tannin. A recombinant form of the arginine kinase enzyme was obtained from the genomic DNA of Trypanosoma brucei brucei by PCR amplification, cloning, expression and His- tag purification. The recombinant arginine kinase of T. brucei (TbAK) was characterised and interacted with Ag/Au nanoparticles or tannin for biochemical characterisation. The interaction between the Ag/Au nanoparticles or tannin and TbAK caused significant decrease in enzyme activity. Further kinetic analysis revealed the interactionswere non-competitive inhibtion, non-spontaneous and involved hydrophobic forces. Fluorimetric analysis suggest that quenching of fluorescence intensity by the nanoparticles or tannin may be static. Available data warrant further kinetic analysis to elucidate other binding properties between the TbAK and the Ag/Au nanoparticles or tannin. The knowledge of such binding properties would aid faster development of selective inhibtors for the TbAK in a drive towards effective chemotherapy for trypanosomiasis.

NANO-62 Nanoparticle / Microparticle - Sensitiser Conjugates as Deeply Penetrating Agents in Photo/Sono-Dynamic Therapy J.F.Callan1, A.P. McHale1, B. McCaughan1, C.Fowley1 and N. Nomikou2

1Department of Pharmacy and Pharmaceutical Sciences, University of Ulster, Coleraine, Northern Ireland. BT52 1SA 2Sonidel Ltd., Dublin 4, Republic of Ireland.

Photodynamic therapy (PDT) is currently approved for use in the UK as a treatment for certain cancerous and pre-cancerous conditions. One obstacle, however, that prevents PDT receiving more widespread clinical appeal is that the light used to activate currently approved photosensitising drugs can only penetrate a few millimetres through human tissue. Therefore, there has been significant activity in this area to improve the tissue penetration capability of PDT. One approach that we have pursued is to utilise the high two photon absorption capability of Quantum Dots (QDs) to indirectly excite a nearby photosensitising drug through an energy transfer

mechanism.1 The advantage of using two-photon irradiation means that longer wavelength light (800nm) can be used resulting in significantly greater tissue penetration (4 times greater at 800nm than 600nm). Another method we are investigating is Sonodynamic therapy (SDT) which uses ultrasound instead of light to activate the sensitiser.2 The benefit of using ultrasound is that it has significantly greater tissue attenuation than ultrasound (several cm’s) thereby enabling access to deeper seated tumours in a minimally invasive manner. In this talk, the use of both QD-sensitiser conjugates and microbubble-sensitiser conjugates will be presented as possible strategies to enable greater tissue penetration in PDT/SDT. 1. C. Fowley, N. Nomikou, A.P. McHale, P. A. McCarron, B. McCaughan and J. F. Callan: J. Mater. Chem. 22, 6456 (2012). 2. N. Nomikou, B. McCaughan, C. Fowley, G. Hamill, A. P. McHale and J.F. Callan: Chem. Commun. 48, 8332 (2012)

NANO-63 Could anisotropic iron oxide nanoparticles lead to further enhancing in magnetic resonance imaging? E. Castro1,2*, J.F. Mano1,2

13B’s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Portugal. 2ICVS/3B’s - PT Government Associate Laboratory, Braga/Guimarães, Portugal.

A magnetic assisted coprecipitation method for the production of anisotropic and isotropic superparamagnetic nanoparticles is presented [1,2]. The preparation of anisotropic iron oxide nanostructures such as needles, rods, whiskers, bars, worms or wires, have attracted much attention and research due to their enhanced optical, electronic, magnetic, and mechanical properties over spherically shaped nanoparticles [3]. The results by XRD,

-potential, DLS, PPMS and MRI showed that no obvious difference on the nanoparticle crystalline structure, while the morphology varied from sphere-like to rod-like and the magnetic properties were significantly changed. It was found that anisotropic iron oxide nanoparticles have a higher degree of crystallinity, relaxivity and saturation magnetization than their spherical counterparts. These novel acicular iron oxide nanoparticles show great promise for use in biomedical imaging applications. [1] Y. Liu, S. Jia, Q. Wu, J. Ran, W. Zhang and S. Wu. Catal. Commun. 12, 717 (2011). [2] W. Zhang, S. Jia, Q. Wu, J. Ran, S. Wu and Y. Liu. Mater Lett. 65, 1973 (2011). [3] C. Yang, J. Wu and Y. Hou. Chem. Commun. 47, 5130 (2011).

NANO-64 Peptide-based Hydrogels for Cell Culture L. Jeannin1, W. Moussa1, M. Bousmanne1, A. Miller2, A. Saiani2, J-B Guilbaud2, L. Szkolar2, C. Pickford2, Y-J. Schneider3, G. Nolleveaux3, A. Joly3, V. Marechal3.

1Peptisyntha, Peptide Biomaterials, Rue de Ransbeek, 310 1120 Brussels, Belgium, 2University of Manchester, Manchester Interdisciplinary Center, Polymers & Peptides research Group, 131 Princess Street, M1 7 DN, Manchester, UK, 3Institute of Life Sciences (group of Cellular, Nutritional & Toxicological Biochemistry) & UCLouvain, 1348 Louvain-La-Neuve, Belgium,

Molecular self-assembly has attracted considerable attention as a means of creating designed materials and devices for nanotechnology. One of the most ubiquitous self-assembly processes in Nature are the hierarchical organization of proteins monomers into long filaments, bundles, and networks. Based on a class of self-assembling oligopeptides originally discovered by Zhang et al. [1], we designed short peptides using an alternating pattern of hydrophobic-hydrophilic amino acids able to form 3D Hydrogels under appropriate conditions. This basic sequence completed by a peptide bioactive sequence and different kind of polymeric parts will allow us to develop at industrial level a technology platform of Soft Materials for Cell Culture and Tissue Engineering. Thanks to their 3D nature and high water content, Hydrogels most resemble to the native extracellular matrix (ECM) that surrounds many types of cells. Moreover, peptide-based Hydrogels ensure by their chemical nature a good cytocompatibility. For these reasons, Hydrogels are regarded as the most promising Soft Materials as new synthetic matrices for Cell Culture. The most recent chemical, analytical and biological results of these Hydrogels will be presented. This Project is partially funded by the Brussels Region (InnovIris – Convention 11-R-154). NANO-65 Plasma Polymerized Poly (ε-Caprolactone)-Poly (ethylene glycole) Copolymer Coatings - As a Biodegradable Carrier for Controlled Drug Delivery Applications S. Bhatt1, J. Pulpytel1, M. Mirshahi2, F.Arefi-Khonsari1

1LGPPTS, UPMC, ENSCP, 11 rue Pierre et Marie Curie, 75231, Paris cedex 05, France 2UMRS 872, CRC, Faculté de Médecine Paris VI, 75006 Paris, France

Catalyst and solvent free low pressure RF plasma polymerization strategy was used to develop multilayered biodegradable PCL-co-PEG (Poly (ε-Caprolactone)-Poly (ethylene glycole) copolymer) coatings [1,2] for the controlled delivery of antineoplastic drug to reduce the nephrotoxicity and myelosuppression. The resulting PCL-co-PEG coatings were characterized by FTIR-ATR and XPS. Multilayered drug delivery devices can be tailored in such a way to have controlled cell-surface interactions and barrier layer dependent release. Cisplatin loaded (118µgm/cm2) PCL-co-PEG coatings with optimized ratios of ε-CL/DEGME monomer feed in the plasma reactor were prepared for controlled cell death applications. MB was used as a model for cisplatin to simulate the drug release kinetics and the nature of which was examined by using the Korsmeyer-Peppas model for polymer coatings. Barrier layer dependent release was investigated by varying the deposition time from 5 to 50 min. Ellipsometric measurements were performed to understand the influence of surface hydration over the different chemically functionalized coatings with the incubation time. Human ovarian carcinoma cells (NIH:OVCAR-3) were cultured in physiological conditions and were seeded in a microplate which was loaded with autoclave coated glass cover slips for different time durations. Cell viability assay was performed to analyze the cell death. The results show that by gradually increasing the barrier layer thickness with deposition time, the amount of MB

released was decreased which was modelled by analysing the MB release kinetics. Investigations of each layer for multilayered PCL-co-PEG coatings were demonstrated over the drug delivery applications in vitro. [1] S.Bhatt, J. Pulpytel, M. Mirshahi, F.A.Khonsari, ACS Macro Lett.,1 (6), 764 (2012) [2] S.Bhatt, J. Pulpytel, M. Mirshahi, F.A.Khonsari, RSC Adv., DOI: 10.1039/C2RA212 11K (2012)

NANO-66 Bactericide studies of Cu-Ag Core-Shell Nanostructures S. Levi1, C. Rousse1, V. Mancier1, S. gangloff2, P. Fricoteaux1

1LISM, EA 4695, UFR Sciences Exactes et Naturelles, BP 1039, 51687 Reims cedex 2, France. 2IPCM, EA 3796, UFR Pharmacie, 51095 Reims cedex, France

Copper and silver are often considered for their bactericides properties. The bactericide applications of these materials under micro and nano structures can be very varied: surface treatment, cleaning solution, packing, smart textile, etc. The elaboration of Cu-Ag core-shell nanostructure offers the opportunity to combine the bactericide properties of these two metals. The proposed synthesis method is cheap and very simple. Nanostructures (spherical particles or nanowires) were obtained by electrolysis of a copper solution (reaction 1) followed by a chemical displacement reaction inside silver solution (reaction 2). The Cu spherical particles were obtained by pulsed sonoelectrochemistry. This method alternates electrodeposition and ultrasonic pulses [1]. The nanoparticles were formed during the electrodeposition phase and were removed by the ultrasounds. The Cu nanowires were prepared using the "template" method [2]. It consists to electrodeposit nanowires inside the nanopores of a polycarbonate membrane. The composition and the reaction kinetics were achieved by X ray diffraction and by energy dispersive analysis. The morphology and the particles size were defined by scanning electronic microscopy, dispersion light scattering and electron energy-loss spectroscopy. At least, the bactericide properties were presented. [1] V. Mancier, C. Rousse, J. Dille, J. Michel and P. Fricoteaux, Ultrasonics Sonochem., 17, 690 (2010). [2] C. Rousse, P. Fricoteaux, J. of materials science, 46, 6046 (2011).

NANO-67 Elaboration of bactericide Cu-Ag Core-Shell Nanostructures S. Levi1, C. Rousse1, V. Mancier1, S. gangloff2, P. Fricoteaux1

1LISM, EA 4695, UFR Sciences Exactes et Naturelles, BP 1039, 51687 Reims cedex 2, France. 2IPCM, EA 3796, UFR Pharmacie, 51095 Reims cedex, France

Some metals (silver, copper, zinc...) exhibit bactericide properties and according to the metal (or oxide) nature, the targeted bacteria differ. The aim of this work is to combine two different bactericide properties in a same structure. Jing et al [1] have shown that this aim could be attempted with a Cu-Ag bi-layer planar structure. Our work extrapolates this idea to Cu-Ag core-shell nanostructures (spherical particles and nanowires). Nanostructures were obtained by electrolysis of a copper solution (reaction 1) followed by a chemical displacement reaction inside silver solution (reaction 2). The Cu spherical particles were obtained by pulsed sonoelectrochemistry. This method alternates electrodeposition and ultrasonic pulses [2]. The nanoparticles were formed during the electrodeposition phase and were removed by the ultrasounds. The Cu nanowires were prepared using the "template" method [3]. It consists to electrodeposit nanowires inside the nanopores of a polycarbonate membrane. The kinetics of the conversion reaction (2) exhibits a great difference according to the

morphology of the nanoparticles. Bactericide properties of these two structures are in course to see if they are also dependent of the morphology. [1] H. Jing, Z. Yu, L. Li, J. of Biomedical Materials Research Part A, 87, 33 (2008). [2] V. Mancier, C. Rousse, J. Dille, J. Michel and P. Fricoteaux, Ultrasonics Sonochem., 17, 690 (2010). [3] C. Rousse, P. Fricoteaux, J. of materials science, 46, 6046 (2011).

NANO-68 Preparation and characterization of poly(D,L-lactic-co-glycolic) acid controlled release nanoparticles containing monoclonal antibody R. Varshochian1, M. Jeddi-Tehrani2, A.R. Mahmoudi2, A. Sabzvari1, F. Atyabi1, R. Dinarvand1

1Nanotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran 2Avicenna Research Institute, ACECR, Tehran, Iran

Rapidly growing application area of antibody-based therapeutics requires novel approach to develop efficient drug delivery systems in which biodegradable polymeric nanoparticles are amongst the best candidates [1]. Bevacizumab is an anti-VEGF monoclonal antibody (mAb) which has been used by ophthalmologists as an off-label drug for treatment of different ocular disease such as choroidal neovascularisation (CNV). Due to its relatively short half-life in the vitreous, repetitive intravitreal injections are necessary to maintain the drug efficiency [2]. To address this disadvantage, mAb loaded PLGA nanoparticles (NPs) have been studied in present research. Inactivation and aggregation of proteins is the major issue in double emulsion encapsulation technique [3]. To overcome this problem 8-10% albumin was added to the internal aqueous phase. VEGF165 binding affinity of bevacizumab was measured by ELISA test which portrays the antibody's bioefficiency. Secondary and tertiary structural stability of antibody was analyzed by circular dichroism (CD) spectroscopy. Ex vivo release of nanoparticles were investigated in diluted rabbit vitreous medium. ELISA and CD spectroscopy results revealed that albumin could preserve bevacizumab activity and structural stability during emulsification procedure and NPs could release active bevacizumab for more than one month. By changing polymer/protein ratio we could improve release manner. Finally it can be concluded that this systematic process could produce efficient sustained release delivery system for antibodies as well as other therapeutic proteins. [1] S. Son, W.R. Lee, Y.K. Joung, M.H. Kwon, Y.S. Kim and K.D. Park, Int. J. Pharm., 368, 178 (2009). [2] P. van Wijngaarden and S.H. Qureshi, Clin. Exp. Optom., 91, 427 (2008). [3] M. van de Weert, W.E. Hennik and W. Jiskoot, Pharm. Res., 17, 1159 (2000).

NANO-69 Glow discharge assisted oxynitriding process of titanium alloy for medical applications T. Wierzchon1, M. Tarnowski1, T. Borowski1, J. Morgiel2, A. Sowińska3, E. Czarnowska3

1Faculty of Materials Science and Engineering, Warsaw University of Technology, PL, 2Institute of Metallurgy and Materials Science, Polish Academy of Science, Cracow PL 3Children’s Memorial Health Institute, Pathology Dept., Warsaw, Poland

Titanium and its alloys are widely used in medicine mainly for the manufacture of implants, plates, bone-fastening screws and prosthesis of the hip joint and medical instruments. In order to improve their resistance to frictional wear, hardness, biocompatibility, as well as to eliminate occurrence of metalosis various methods of surface engineering are successfully applied [1,2].The presented paper shows an oxynitrided diffusive layers

produced on Ti6Al4V titanium alloy under low-temperature glow discharge oxynitriding process. The produced surface layers is TiO2-TiN+Ti2N+αTi(N) type with porous or nonporous titanium oxide zone of thickness in range of 100-300 nm dependly on process parameters. Its microstructure (analyzed by TEM and SEM), chemical and phase composition (analyzed by EDS and XRD) and topography (analyzed by optical profilometer) are presented. This oxynitrided surface layer exhibits hardness about 1020 HV0.05. Its wear resistance tested by the “three rollers and taper” method (modified “four ball method”) increases significantly in comparison to reference titanium alloy. Biological properties were investigated in in-vitro tests with human platelets and osteoblast-like cells. Nanocrystalline TiO2surface layer without porous shows better antithrombogenicproperties than titanium alloy, while TiO2 with nanoporous structure characterizes better osteoblasts adhesion and proliferation than Ti6Al4V alloy. Therefore oxynitrided surface layers can widen significantly application range of titanium and its alloys for medical application as e.g. cardiovascular and bone implants. [1] X. Lim, P. K. Ch. Ding, Materials Science and Engineering R47 (2004) 49 [2] E. Czarnowska, et al., Journal of Nanoscience and Nanotechnology 11 (2011) 8917

NANO-70 Modelling Polymersome Delivery of Anticancer Therapeutics S. Webb1, I. Sorrell1, R.J. Shipley2, G. Battaglia3, V. Hearnden4, H. Colley4, C. Murdoch4

1Department of Molecular and Clinical Pharmacology, University of Liverpool. 2Department of Mechanical Engineering, University College London. 3Department of Biomedical Science, University of Sheffield. 4Department of Clinical Dentistry, University of Sheffield.

Polymersomes are synthetic nano-spheres that have the potential to specifically deliver anticancer therapeutics to tumor cells. They are an ideal delivery vector because of their nanometer size, structural architecture, stealth characteristics and the ability to attach tumor-specific ligands to their outer surface. The possible benefits of targeted polymersome drug delivery include reduced off-target toxic effects in healthy tissue and increased drug uptake by diseased tissue. In this study, in vitro experimentation is used to validate a mathematical description of polymersome-mediated therapy. Mathematical predictions are given for how the properties of polymersomes (size, membrane properties) affect rates of polymersome internalisation by tumor cells and therefore treatment efficacy. This allows for an optimisation of treatment design prior to clinical development. A key focus is polymersome-drug delivery via the clathrin-mediated endocytotic pathway. We include in our theoretical description specific details of this process (including specific binding ligands, receptor clustering, ligand tether length) and show how the characteristics of receptor targeting, number of ligands per polymersome, and polymersome concentration alters its uptake by tumor cells. The theoretical model is validated against in vitro studies using polymersome drug delivery to cell monolayers derived from different tumors. Next we describe how to scale-up the model to a tissue representation of the tumor mass. In this description the concentration of solutes (including oxygen and polymersomes) are tracked throughout the tumor using computational models based on realistic tumor vascular network structures with associated blood flow that have been constructed from confocal microscopic observations of a xenograft sub-cutaneously implanted tumor. Our results quantify the dependence of polymersome uptake rate and tumor distribution on polymersome properties and dosage. This multiscale approach identifies the key polymersome properties that influence treatment outcome and enables the treatment regime to be designed specifically in response to these features.

NANO-71 Dual decorated liposomes with MAb OX-26 and ApoE3 peptide derivative for brain targeting. K. Papadia1, E. Markoutsa1, A. Cagnotto2, M. Salmona2, S G. Antimisiaris 1,3

1Lab Pharm Technology Department of Pharmacy, University of Patras, Rio-Greece 2Mario Negri Institute, Milan, IT. 3FORTH/ICES, Rio-Greece

Dually decorated liposomes (dd-LIPs) were prepared, with Ox-26 monoclonal antibody against transferrin receptor (TfR) and peptide analogue of Apolipoprotein Ε3 (ApoE3) on their surface, in order to investigate potential additive/synergistic effect on BBB transport efficiency. hCMEC/D3 cells over-express TfR and LDL receptors, and were used as an in vitro (cell culture) model of BBB. LIP surface was decorated with biotin-streptavidin ligation for Ox-26 and maleimide-cysteine ligation for ApoE3 peptide. Liposomes were characterized for ligand attachment, size, charge and stability (in serum). Mean diameter of LIP was always between 150-200 nm. LIP with both ligands (ApoE3+Ox26) had substantially higher uptake by hCMEC/D3 cells, compared to controls (pegylated LIPs or LIPs with one ligand). In monolayer experiments, ApoE3-LIPs or Ox26-LIPs equally permeate the BBB, while dd-LIPs demonstrate the highest permeation percent (additive effect for BBB transport). In order to confirm transport of intact LIPs, FITC-dextran (hydrophilic) and RHO-lipid (lipophilic) were used as labels and the transport of both labels was followed; and found to be practically equal. Lucifer yellow, was used a hydrophilic paracellular transport label. MTT (cytotoxicity) studies were also performed. It was proven that these dd-LIPs are not toxic and that their transport across the cellular model of BBB is not influenced by paracellular effects. The research has received funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 212043. NANO-72 Controllable Synthesis, Characterization and Antibacterial Activity of Sandwich-Like Graphene Oxide-Rare Earth Complex Nanocomposites Dr. X.F. Yang1, Dr. H.Y. Cui2, J.L. Qin1 1School of Materials Science and Engineering, Jiangsu University, Zhenjiang, 212013, P. R. China. 2School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, P. R. China.

With the rapid development of synthesis and functionalization approaches, graphene and its related derivatives have shown outstanding potentials in many fields. Among various applications, the biological & biomedical application of graphene-based materials is a relatively new area with significant potential and has attracted ever-increasing interests over the past few years. Many efforts have been made to explore the use of graphene-based materials for widespread applications including drug/gene delivery, cancer therapy, antibacterial materials, biosensing and imaging. Rare earth (RE) complexes have been proved to be one of promising candidates for the preparation of antibacterial materials, however, for practical applications, there are several problems that need to be resolved including poor solubility/dispersibility in aqueous solution, relatively higher cell cytotoxicity to mammalian cells and the high cost of raw materials. Chemical exfoliated graphene oxide (GO) sheets possess many reactive hydrophilic oxygen-containing groups and large specific surface area, superior dispersibility of GO sheets in aqueous solution makes them excellent building blocks for the generation of novel antibacterial hybrid materials through the incorporation of various kinds of antibacterial materials. Herein, we present a facile and effective method to prepare GO-RE complex nanocomposites, it is believed that the electrostatically-driven assembly of positively charged RE ions on the negatively charged GO sheets, as well as the nucleation and in-situ growth of RE complexes on GO sheets favors the generation of well-defined sandwich-like GO-RE complex

nanocomposites. The structure, composition, size and morphology of as-synthesized nanocomposites are fully characterized. Three methods including inhibition zone, minimum inhibitory concentration (MIC)/minimum bactericidal concentration (MBC), time-dependent bactericidal activity calculated by residual bacteria count were employed to investigate antibacterial performance of nanocomposites against seven types of bacteria. Our findings show broad-spectrum antibacterial activity of nanocomposites that possibly caused by synergistic antibacterial effects between GO sheets and formed RE complexes. Due to characteristics of as-synthesized GO-RE nanocomposites, with the advance in synthetic procedures of chemical exfoliated GO sheets, our findings may have implications in the the low-cost, large-scale production of antibacterial materials for potential photocatalytic applications in different fields. [1] X.D. Wang, N.L. Zhou, J. Yuan, W.Y. Wang, Y.D. Tang, C.Y. Lu, et al., J. Mater. Chem., 22, 1673 (2012). [2] S. B. Liu, T.H. Zeng, M. Hofmann, E. Burcombe, J. Wei, R.R. Jiang, et al., ACS Nano, 5, 6971 (2011). [3] O. Akhavan, E. Ghaderi, ACS Nano, 4, 5731 (2010). [4] W. B. Hu, C. Peng, W.J. Luo, M. Lv, X.M. Li, D. Li, et al., ACS Nano, 4, 4317 (2010).

NANO-73 Effect of substrate temperature on properties of Bi-2212 thin films using rf magnetron sputtering technique

Nguyen Thi Mua1,2, Sudaresan2, C.N.R.Rao2

1 Police Department of Fire prevention Fire Fighting and Rescue, Ministry of Public Security,No 2 Dinh Le, Hoan Kiem, Hanoi 2 .Jawaharlal Nehru Centre for Advanced Scientific Research. Bangalore 560 064, India

We were prepared superconducting Bi-2212 thin films on MgO (100) substrate single crystal using rf-magnetron sputtering. A series of thin made annealed in chamber with environment oxygen .Target Bi-2212 have been use in this technique. The films were annealed for different period of time in oxygen at 1atm and the films were deposited at different temperature substrate. Effect of annealing time and the temperature of the substrate on the quality and the phase formation of Bi-2212 thin films have studied. The influence of substrate temperature also has been investigated by X-ray diffraction, scanning electron microscopy (SEM), resistivity and ac -susceptibility. It was found have get the good Bi-2212 thin films with condition was annealed in environment oxygen at 1atm C for 4 hours and deposited at 6000 C substrate temperature. It is well that, there are several phases in the BSCCO system. The composition of each phase is expressed by general formula of Bi2Sr2Can-1CunOx ( n=1,2,3) and Tc=10,85,110K respectively. Here after, we used the abbreviation 2201, 2212, 2223 phases. In spite of this potential of BSCCO materials, most application of high- Tc superconducting thin films are based on YBCO materials, since the in situ growth of BSCCO thin films is comparatively difficult. The difficulties are due to the fact that, superconducting properties are strong dependent on variations of stoichiometry. For the deposition of BSCCO thin films additional complexity arises from possible phase mixture and oxygen over doping. Different technique such as molecular beam epitaxy ( MBE) [1,3]], metal organic chemical vapor deposition (MOCVD) [4,5], sputtering, pulsed laser deposition ( PLD) etc…have been employed. Nonetheless, the few application of this BSCCO thin films system may be due to the difficulty of obtaining high- quality films composed of the respective Bi22(n-1)n pure phases. The phase diagram of the three stable phases occasionally occurs during thin films fabrication or single crystal growth. Accordingly, it is indispensable to establish thin films. The application of high-Tc superconductors is likely to be in the form of thick and thin films in microwave devices, bolometer, flux transformers, magnetic shielding devices, etc. In the application of superconducting films, the substrate plays a vital role. The chemical reactivity between the substrate and the superconductor at the processing temperature is the most crucial factor for obtaining a high-Tc superconductor films. In addition, the substrate should not poison the superconductor and deteriorate its properties [1,2]. MgO is one of the useful substrate for designing several devices such as filter and a mixer for high frequency because of the low dielectric constant, low dielectric dissipation factor and low price. Whereas, MgO is regarded as a worst substrate in popular ones due to a large lattice mismatch of about 10% in constants to the a- (or b-) lattice constant in the Bi

superconducting structure. The difference in the lattice constant and the thermal expansion coefficient between MgO and HTSC materials influence directly on the thin films quality. In this paper, we report on the preparation of target on the deposition Bi-2212 phase thin films, using in situ rf-magnetron sputtering technique. We discuss the superconducting properties and the fabrication conditions for single-phase and mixed- crystalline BSCCO thin films. We used single powder target with various compositions of Bi:Sr:Ca:Cu=2:2:1:2. The target were prepared from the starting materials of Bi2O3, SrCO3, CaCO3 and CuO by grinding, mixing and sintering at 8300C for 24 h in air. Each target was into a disk with a diameter of 50 mm and thickness of 2 mm . The sputtering gas a mixture of Ar and O2, and the films was deposition on the polished MgO (100) substrate. The ratio of Ar gas pressure to total gas pressure was fixed at the optimum value Ar: O2=4:1. The sputtering time and rf power were about 2-4 hours and 40W, respectively, and the film thickness was estimated to be approximately 350 nm. The substrate temperature was changed from 5180C to 6000C. The target -substrate distance is 75 mm. In order to improve the superconducting properties, we carried out in situ annealing for Bi-2212 thin films. After deposition, the films were in situ annealing was carried out for the period from 3h to 5 h in flowing O2 gas with PO2=1 atm. The crystal structure and the surface morphology of the thin films was investigated by X-ray diffraction ( XRD), and scanning electronic microscopy ( SEM) the crystalline of the films was estimated from the

full-width at half -maximum (FWHM) value of the -2 scan. The atomic compositional ratio of the deposited films were examined by energy dispersive X ray spectrometry ( EDX). The temperature- resistivity characteristics of thin films were measured from 300K to 5K by using the standard four-point probe method using silver paste contacts. Patterns of the samples were deposited at different temperature substrate. It is founds that the x-ray reflection lines became broader and their peak positions shifted significantly, as expected from the size effect of x ray diffraction from a layered modulated structure. The X-ray patterns of the films prepared at different substrate temperature, keeping the total gas pressure and the Ar/O2 ratio constant. In this figure, all strong diffraction lines assigned of 2212 phase. But, as one can see clearly from the pattern, the proportions of 2212 phase will increase as the substrate temperature increase. In X-ray diffraction, the best films obtained show only 2212 (00l) peak. Although the FWHM of the rocking curve is only 0.80. Only a single peak actually appears at the midpoint between the individual Bi-2201 and Bi-2212 peak. This means that the two phases are not separated in

macroscopic domains larger than the X-xay penetration of 1m, and that the intergrowth takes place in the domain due to stacking modulation of the Ca and CuO2 layer between the Bi-2201 and Bi-2212 phases. SEM images of the surfaces of thin films. We observe the films surface are almost smooth and homogeneous. As the temperature of the substrate increase, the formation of large grain become more favorable and we have a better link between the grains. Further investigation is necessary and to determine the exact experimental parameters and to reveal the mechanism of the ion irradiations influence on the Bi-2212 film characteristics. The result of resistivity measurement for the sample prepared in situ method . Thickness of the films is about 350 nm. As show in Fig.3, the superconducting transition was apparent in films at 5180C, 5250C, 5670C and 6000C substrate temperature. The superconducting properties are strongly affected by the substrate temperature during deposition. The transitions was, however, relatively gradual; that is the Tc, on-set was about 84K and the Tc, R=0 was about 70K for films deposited at 6000C substrate temperature, and 80K and 66K for films deposited at 5180C substrate temperature. In this figure, the samples show superconducting transitions, and it have a metallic behavior in their normal state. The T c, onset was about 84 K and the Tc,R=0 was about 70K for films was deposited at 6000C substrate temperature and annealed within 4 hours at chamber with environment oxygen with P02= 1 atm. It has been report that Bi-2212 films could be formed in situ at substrate temperature 6000C by rf magnetron sputtering. In this work, the optimal conditions for synthesis of Bi-2212 films with good crystallographic properties were found to be 6000. In this research, we investigated the phase intergrowth in BSCCO thin films. The process formation of the Bi-2212 superconducting phase in in situ rf magnetron sputtering thin films has been studied, and a reproducible fabrication procedure leading on almost single 2212 phase BSCCO compound was achieved, using MgO (100) as a substrate. The fabrication films were highly oriented with c- axis perpendicular to the surface. Structural studies, based on the X-ray diffraction measurements. The result of our studied show. It was found have get the good Bi-2212 thin films with condition was annealed in environment oxygen at 1 atm C for 4 hours and deposited at 6000 C substrate temperature. We would like to

thank Prof. C.N.R.Rao, Dr. Sundaresan, Laboratory Superconductivity and Magnetism, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore 560 064, India.

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NANO-74 Engineering the Surface of POSS-PCU Nanocomposite by Plasma to Improve Blood Compatibility A. Solouk1, B.G. Cousin2, H. Mirzadeh1, A.M. Seifalian2,3

1 Polymer Engineering Faculty, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran

2Centre for Nanotechnology and Regenerative Medicine, Division of Surgery and Interventional Science, University College London (UCL), London, UK 3Royal Free Hampstead NHS Trust Hospital, London, UK

A novel nanocomposite polymer comprising polyhedral oligomeric silsesquioxane (POSS) nanoparticle and poly (carbonate urea) urethane (PCU) polymer was developed for biomedical applications1. The polymer has in vitro and three years large animal studies in term of biocompatibility. It is widely accepted that most important biological interactions happen on the surface of a biomaterial. Hence, surface modification of biomaterials has become an interesting topic in medical engineering as well. Among various surface modification techniques, plasma surface modification (PSM) has been proven to be very effective 2. Actually, cells in their in vivo microenvironment constantly encounter and respond to a multitude of signals including biochemical or biophysical cues. While the role of biochemical signals such as growth factors and cell culture medium has long been appreciated, the importance of biophysical signals like surface topography has only recently been investigated with the progress in microfabrication technologies3. In fact, the microfabrication technologies have allowed for the generation of biomaterials with microscale topographies and give the opportunity to study the effect of topography as one of the most important biophysical cues at the cell-substrate interface3. In this study, using PSM topographies with different geometries was produced on the surface of POSS-PCU nanocomposite and the varying in their blood compatibility was evaluated. Sheets (40 × 40 mm) of POSS-NC, 100 µm thickness, were manufactured and were put in plasma reactor chamber (Nano-RF-PC, Diener, Germany) which was evacuated to 2 x 10-1 mbar, and pretreated with oxygen plasma in different times and powers. Untreated film was used as control. The effect of oxygen plasma treatment on the surface topography was monitored using atomic force microscopy (AFM). To test cell response to the topographical changes on the surface, human umbilical vein endothelial cells (HUVECs) were cultured on the substrates with different amount of roughness and also platelet adhesion were assayed using platelet-rich plasma (PRP). AFM three-dimensional (3D) surface topography images of O2 – plasma-treated POSS–PCU films showed formation of porous structures and pronounced elevated features composed of POSS within the polymeric matrix as shown in Fig 1. It was found that the surface roughness of the films decreased as a result of oxygen plasma treatment. It was also observed that the adhesion, coverage, and growth of HUVECs rose significantly in number (Fig. 2) simultaneous with decrease in the amount of adhered platelets. These findings suggest that oxygen plasma treatment of POSS-PCU could be an attractive way to improve its blood compatibility. Outcome will be higher patency rate of small-diameter vascular grafts.

[1] A. Solouk, B. G. Cousin, H. Mirzadeh, M. Solati-Hashtjin, S.Najarian, and A. M. Seifalian. Biotechnol. Appl. Biochem.58,147, (2011). [2] A. Solouk, B. G. Cousin, H. Mirzadeh, A. M. Seifalian, Biotechnol. Appl. Biochem. 58, 311, (2011). [3] M. Nikkhah, F. Edalat, S. Manoucheri, A. Khademhosseini. Biomater. 33, 5230, (2012).

NANO-75 Cell-to-cell adhesion and adhesion to extracellular matrix molecules in directing stem cell proliferation and differentiation Y.L. Ng1 and L. Buttery2

1Department of Chemical and Environmental Engineering, University of Nottingham, University Park, Nottingham, UK, 2Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK

The precise control of stem cell proliferation and differentiation using appropriate cell signalling is of paramount importance for the production of cells and tissues for therapeutic applications. Formation of embryoid bodies widely practiced as part of the protocol for differentiating embryonic stem cells (ESCs) was shown here to enable non cell-specific contact growth arrest where the expansion ratio of ESCs was reduced from more than 17 times in cells that were plated on extracellular matrices (ECMs), to 1.46 to 2 times in cells that were aggregated. In addition to dexamethasone, ascorbate and beta-glycerolphosphate (DAG), adhesion to insoluble collagen and fibronectin was shown to be vital for osteogenic differentiation and the production of pure cultures of osteoblasts that mineralised. On the contrary, cell aggregation and experimental controls of plating on gelatine and poly-D-lysine (PDL), and feeding with the soluble forms of collagen, fibronectin, gelatine and PDL were shown to produce mixed populations of osteoblasts and cardiomyocytes. The role of ECM in stem cell differentiation was further demonstrated with neural stem cells where laminin alone was sufficient to differentiate the cells. Results from this study showed the importance of incorporating specific cell-to-cell and cell-to-ECM signalling in bioprocessing for the rapid production of pure cultures of specific cell types. NANO-76 Morphology controlled porous calcium phosphate nanoplates and nanorods with enhanced protein loading and release functionality P.J. Reardon1,2, J. Tang1, J.Huang2 1 Department of Chemical Engineering, UCL, London, WC1E 7JE, UK. 2 Department of Mechanical Engineering, UCL, London, WC1E 7JE, UK.

As a robust and biocompatible material for drug delivery, calcium phosphate nanoplates and nanorods with controllable pores are first synthesized without the use of any toxic surfactants by an energy efficient microwave assisted process. Correlation between synthesis temperature and the prepared samples’ characteristics is investigated with the aim of controlling material morphology and then textural properties, resulting in control over protein loading and release performance. Using a typical representative protein, bovine serum albumin (BSA), loading has been found to be the highest for the nanoplate samples synthesized at 200 °C. More importantly, the loading efficiency is 2.5 times higher than that of a commercial hydroxyapatite nanomaterial even if the former has approximately half the surface area of the latter. Furthermore, the delivery period is different from the commercial hydroxyapatite nanomaterial, exhibiting two delivery stages (fast and slow stages) and lasting 8 times longer, indicating the morphology controlled nanostructured calcium phosphate could be a promising candidate for drug delivery with increased loading capacity and longer release period.

NANO-77 Silk Scaffolds for Cancer Model Engineering E. Dondajewska1, K.Kazmierska2, A. Mackiewicz1,3, H. Dams-Kozlowska1,3

1 Department of Cancer Immunology, Poznan University of Medical Sciences, Poland 2 Poznan NanoBioMedical Center, Adam Mickiewicz University in Poznan, Poland 3 Department of Cancer Diagnostics and Immunology, Greater Poland Cancer Centre, Poznan, Poland

For almost a century two dimensional (2D) cell culture has been one of the most basic techniques in biology and medicine. Recently, it has proven to be inadequate for modern studies. Scientists appreciate the superiority of a three-dimensional (3D) cell culture, since it closely mimics the cell attachment, cell-cell interactions and extracellular matrix formation. Thanks to its biocompatibility, excellent mechanical properties and slow biodegradability, silkworm silk is a perfect biomaterial for cell studies and in vivo experiments. Moreover, self-assembling property of silk allows shaping it into various structural forms like scaffolds, fibers, films what can be useful for cell culture methods. The aim of the study is to engineer the heterologous 3D cancer model. It comprises of analysis of adhesion and growth of breast cancer cells and fibroblast on silk scaffolds. 6.7% aqueous B. mori silkworm silk solution was used to prepare the scaffolds by salt leaching method. Silk scaffolds of 250-500 µm pore size were formed. Two cell lines: EMT6 – murine breast cancer cells, and 3T3 NIH – murine fibroblasts were genetically modified by lentiviral vector to express the fluorescent proteins TurboFP635 and GFP, respectively. Cells were cultured separate as well as in a co-culture. 2D and 3D cultures were compared. Cells were observed using fluorescent, confocal laser, and scanning electron microscopy. The microscopic study of cell-seeded silk scaffolds indicated very good seeding and expansion of both cell lines. There was difference in cell morphology between cells cultured in 2D and 3D environment. Moreover, expansion of EMT 6 cells was much higher than fibroblasts. Silkworm silk scaffolds have proven to be good matrices for cell culture. The heterologous 3D cancer model composed of cells like epithelial, fibroblast and cancer will provide the microenvironment closely related to one observed in vivo and can be used for the cytotoxicity testing of drugs and nanoparticles. NANO-78 Site-specific drug delivery using biodegradable “nanoparticle-in-membrane” systems S. GuhaSarkar, R. Banerjee Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai, India.

The use of nanotechnology in cancer therapy has shown immense potential to significantly enhance the scope and efficacy of current cancer treatment modalities. Apart from systemic administration, there is a need for regional drug delivery systems that may be able to improve site-specific drug localization, and show potential use as adjunct therapy and for post-operative management of tumors. We have developed “nanoparticles-in-gel” systems utilizing drug-loaded lipid nanocarriers incorporated into biopolymeric membranes, such that the nanoparticles provide sustained release of the encapsulated drug, while the membrane allows localized administration in the form of transdermal patches and intra-tumoral inserts. Such a bio-conjugate system may also allow dual delivery of drugs, with hydrophobic drugs delivered via the lipid nanoparticles and hydrophilic moieties incorporated into the membrane matrix. The liposomes have sizes 121 ± 2 nm and show a sustained release of paclitaxel over 24 hours. Biophysical studies with Langmuir Blodgett monolayers of rigidly packed DPPC lipids showed an increase of 13.1 ± 1 mN/m in surface pressure (Δπ) in the presence of liposomes, indicating perturbation of the rigid monolayer and suggesting that the liposomes may enhance penetration through the lipid lamellae in the stratum corneum. In vitro studies have shown a high degree of uptake of the

liposomes by L929 fibroblast cells, and high cytotoxicity to MDAMB-231 breast cancer cells, with an IC50 of 300nM. Biocompatibility studies with the liposomes and the membrane showed they had no toxic effects. In vivo penetration studies with mouse skin showed a 1.5-fold enhancement of average fluorescence intensity and area of dye penetration in the nanoparticle-in-membrane systems, compared to dye-loaded liposomes alone. In vivo efficacy studies are in progress. Our initial studies with nanoparticles-in-membrane systems have shown feasibility for use as transdermal patches and subcutaneous membrane inserts and may serve as useful adjuncts for post-operative treatment in breast cancer. NANO-79 Metallic Hybrid Nanoparticles For Image Guided Drug Delivery C. Barnett1, M. Gueorguieva2, M. Lees3, R. Darton1, D. McGarvey 1, C. Hoskins1

1 Institute of Science and Technology in Medicine, Keele University, UK. 2 Institute of Medical Science and Technology, University of Dundee, UK. 3 Superconductivity and Magnetism Group, University of Warwick, UK.

Hybrid nanoparticles formed from iron oxide cores surrounded by gold nano-shells are becoming increasingly applicable in biomedicine [1-2]. Recently, their ability to act as MRI contrast agents and nano-heaters was reported [3]. However, little is known of their stability, biocompatibility, cellular interaction or ability to deliver drug molecules. Here we synthesised Fe3O4, coated with gold and surface coated with poly(ethylene glycol). The physical stability and effect in vitro on human BxPC-3 and differentiated U937 cell lines for ROS and LPO production, LDH leakage and their cytotoxic effect was determined. Additionally, model drug 6-Thiguanine (6-TG) was conjugated to the particle surface. 6-TG is used clinically to treat leukaemia. Here the ability of these particles to act as a multifunctional chemotherapy was eluded. UV-Vis and TEM confirmed nanoparticle formation with particles being 70 nm in diameter. The nanoparticles were observed to be stable in physiological media over a 2 week period and did not show any cytotoxic effect or other adverse cellular response after incubation. After 6-TG conjugation the hydrodynamic radius increased from 150 nm to 230 nm. The concentration ratio of iron/gold/drug was 3/1/10 were in every 3mg of Fe3O4, 1mg of Au and 10 mg of 6-TG was present. 6-TG-nanoparticle increased the cellular uptake after 4h and IC50 after 24 h compared with free drug, 21-fold and 10-fold respectively in BxPC-3 cells. Further work is on-going to fully exploit the properties of these nanoparticles for thermo responsive drug delivery. [1] I.Y. Goon, L.M.H. Lai, M. Lim, P. Munroe, J.J. Gooding, R. Amal, Chem. Mater,. 21, 673-681 (2009). [2] E.D. Smolensky, M.C. Neary, Y. Zhou, T.S. Berquo, V.C. Pierre, Chem. Comm., 47, 2149-2151 (2011). [3] C. Barnett, M. Gueorguieva, M. Lees, R. Darton, D. McGarvey, C. Hoskins, J. Nanopart. Res., In press.

NANO-80 Remotely triggered scaffolds for controlled degradation in biomedicine P. Roach, D. McGarvey, C. Hoskins

Institute of Science and Technology in Medicine, Keele University, UK.

Scaffolds fabricated for tissue engineering purposes require controlled pore structure and size, allowing diffusion of nutrients and toxins to in-growing tissue. Biodegradable scaffolds that collapse after a fixed time period, either naturally or after external stimulus, offer further control for long term use. Fe3O4-Au hybrid nanoparticles (HNPs) have shown increasing potential for biomedical applications such as image guided stimuli responsive drug delivery. Here we demonstrate the incorporation of HNPs into a biocompatible scaffold in order to allow an

(C)

(E) (F)

external switch on scaffold characteristics. Using a laser to irradiate the particles in situ, a very localised heating effect causes thermally-responsive scaffold materials to deform mechanically. Additionally, growth factors or drug molecules can be incorporated, being thermally released into the surrounding tissues upon irradiation. In conjunction, the dense nanoparticle cores will also provide contrast in MRI imaging which can be used to visualize and monitor scaffold position and degradation. Here we successfully fabricated biodegradable scaffolds based on thermo-responsive poly(N-isopropylacrylamide) (NipAM) polymers. Nanoparticles acting as the trigger for controlled degradation and release were synthesised using a wet chemical precipitation method followed by electrochemical gold coating, and incorporated into the intrinsic structure of the scaffold. The constructs were fully characterised using TEM, SEM, DSC, PCS, FTIR and microscopy. We investigate the potential of such scaffolds as thermally triggered systems using a Q-switched Nd:YAG laser. These studies show that incorporation of HNPs result in scaffold deformation after irradiation due to the internal structural heating. Biological safety was determined using live/dead fluorescence staining and cellular stress levels deduced via ROS and LPO quantification. The constituents did not show any cytotoxic effect or adverse cellular response on exposure to 7F2 cells. Our data highlights the potential of these hybrid-scaffold constructs for regenerative medicine. Further work is on-going to further exploit these unique properties for controlled drug delivery. NANO-81 Functionalized spider silk proteins for targeted cancer therapy A. Florczak1,2, K. Kazmierska1,2, C. Kieda3, V. Filas4, A. Mackiewicz2,5, H.Dams-Kozlowska2,5

1 NanoBioMedical Centre, Faculty of Physics, Adam Mickiewicz University, Poznan, Poland 2 Chair of Medical Biotechnology, Poznan University of Medical Sciences, Poland 3 Centre for Molecular Biophysics, National Centre for Scientific Research, Orleans, France 4 Department of Cancer Pathology, Greater Poland Cancer Centre, Poznan, Poland 5 Department of Cancer Diagnostics and Immunology, Greater Poland Cancer Centre, Poznan, Poland Spider silks exhibit remarkable properties such as mechanical strength, toughness, biocompatibility and biodegradability, which make them excellent materials for a wide range of biomedical applications. Bioengineered spider silk proteins are based on consensus motifs of natural spider silks and hybrid spider proteins combines the bioengineered silk domains, which are responsible for biomaterial structure with polypeptide sequence which allows functionalization of the silk biomaterial. Cancer cell binding silks which self-assemble into micro- and nanospheres may offer potential for the development of targeted drug delivery systems for cancer treatment. The synthetic genes of silk MS1 (15mer composed of 15 repeats of consensus sequence MaSp1 derived from N. clavipes) and its hybrid variants H2.1MS1, H2.2MS1, MS1H2.1 and MS1H2.2 (MS1 fused to Her2 receptor binding domains H2.1 and H2.2) were constructed. Bioengineered silk productions were carried out in E. coli system and proteins were purified using thermal extraction method. The soluble form of proteins, silk-bead conjugates and silk spheres were tested in Her2 receptor binding assays. The binding efficiency was analyzed by dot-blot, flow cytometry, fluorescent and confocal microscopy. The presence of Her2 receptor on SKOV3, SKBR3 and MSU1.1 was examined by immunohistochemistry. The functionalized proteins indicated considerably higher binding properties to the Her2 overexpressing cells when compared to Her-2 receptor-free cells and comparing with the control protein without the functional domain. The preliminary results demonstrate the potential of bioengineered hybrid silk proteins as a new family of highly tailored systems for cancer therapy.

NANO-82 Bimodal Contrast Agents for the in Vivo Imaging: The Elaboration of Functionalized SuperParamagnetic Iron Oxide Nanoparticles J. Paris1, J. Boudon1, Y. Bernhard2, R. Decréau2, R. Mayap Talom1, L. Maurizi1, F. Bouyer1, P. Walker3, L. Dumont4, D. Vandroux4, F. Denat2 and N. Millot1,* 1 Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 6303 CNRS/Université de Bourgogne, 21078 Dijon cedex, France 2 Institut de Chimie Moléculaire de l’Université de Bourgogne, UMR 6302 CNRS/Université de Bourgogne, France 3 Service de Spectroscopie par Résonance Magnétique, Centre Hospitalier de Dijon, 21033 Dijon, France 4 Société NVH Medicinal, 57 Rue Vannerie, 21000 Dijon, France

Nowadays, nanotechnology offers a large field of applications in medicine and biology [1, 2]. This domain is very promising in particular regarding the detection and the treatment of pathologies. SuperParamagnetic Iron Oxide Nanoparticles (SPIONs) represent some of the most important nanoparticles used as MRI contrast agents [3] or for hyperthermia treatment [4]. Recently, our group has developed continuous hydrothermal syntheses, in subcritical and supercritical water conditions. The nanoparticle size is smaller and their size distribution is narrower than those of powders obtained with other common methods. Moreover, citrate-stabilized SPIONs were obtained in a continuous way [5]. In this study, we present the development of multimodal contrast agents based on functionalized SPIONs for MRI, SPECT, PET and optical imaging (OI). At first SPIONs are prepared by the grafting of organic functions (NH2, COOH or SH) to allow the implementation of more specific organic molecules among them: macrocyclic chelating agent for nuclear imaging (DOTA) [6] or phthalocyanines derivatives (for fluorescence and photodynamic therapy) [7]. Second, a covalent coupling of functionalized PEG is necessary to ensure the biocompatibility and the stability of these nanoparticles [8]. The grafting of the organic molecules (DOTA, PEG and phthalocyanines) was proven by different analyses such as fluorescence, UV-vis spectra, XPS, TGA and elemental analysis [9]. This resulted in novel bimodal contrast agents detectable for both MRI and near infrared optical imaging but also for MRI and SPECT (or TEP) imaging. The first results concerning the in vivo biodistribution show a reduced capture of PEGylated SPIONs by mice liver and suggest that our contrast agents freely circulate longer in mice compared to a commercial product tested. MTT assays prove that these iron oxide suspensions have no apparent cytotoxicity on the different cell lines tested (cardiomyocytes, macrophages and hepatic cells). [1] A. Schroeder, D. A. Heller et al., Nat Rev Cancer, 12, 39 (2012). [2] W. J. Stark et al., Angew. Chem. Int. Ed. 50, 1242 (2011). [3] Y.-X. Wang, S. Hussain et al., Eur. Radiol. 11, 2319 (2001). [4] M. Johannsen, U. Gneveckow et al., Eur. Urol. 52, 1653 (2007). [5] L. Maurizi, F. Bouyer et al., Chem. Commun. 25, 8857 (2011). [6] C. Bernhard, C. Goze et al., Chem. Commun. 46, 8267 (2010). [7] S. Gundy, W. Van der Putten et al., Phys. Med. Biol. 49, 359 (2004). [8] L. Maurizi, H. Bisht et al., Langmuir 25, 8857 (2009). [9] J. Boudon, J. Paris, et al. in preparation

NANO-83 Biologically Active Nanoparticulate Based Injectable Polymeric Matrices for Tissue Regeneration D. Dyondi1, T. J. Webster2, R. Banerjee1

1 Indian Institute of Technology Bombay, Powai, Mumbai, Maharashtra, 400076, India 2 Brown University, Providence, Rhode Island, 02906, USA

Nanomaterials have been extensively utilized in tissue engineering and drug delivery with the aim of mimicking nano- and micro-structure of native tissue. In situ gelling systems involving natural polymers to obtain injectable biodegradable scaffolds is emerging as an alternative to tissue regeneration that can eliminate the need for an additional surgery [1]. The current study describes one such system using gellan gum along with xanthan entrapping growth promoting agents and precursor cells- chondrocytes for tissue regeneration of cartilage. We developed a nanoparticulate based system for dual growth factor delivery. The hydrogels have been well characterized for gelling times, viscoelasticity, syneresis, previously by the group [2]. Growth factors, IGF I and PDGF BB were adsorbed over solid lipid nanoparticles nanoparticles (SLN), with the hydrogel matrix and SLN as the two matrices. Entrapment of growth factors in nanoparticles within the hydrogels provides a dual barrier hence a slower release profile keeping in view the temporal requirement of different growth promoting agents at different stages of tissue regeneration. Cell viability was tested with L929 cells, pig chondrocytes and human chondrocytes. Live cells within hydrogels were visualized using live dead staining (Calcein AM and propidium iodide) at day 7. 28-days differentiation study showed an increase in the DNA and protein content in growth factor loaded gels as compared to only hydrogels. Higher glycosaminoglycan (GAG) content was observed for growth factor loaded gels as estimated by DMMB assay. Also, dual growth factor loaded gels showed a higher DNA, total protein and GAG as compared to single growth factor loaded gels. [1] J.D. Kretlow, L. Klouda and A.G. Mikos, Adv. Drug Deliv. Rev., 59, 263 (2007) [2] D. Dyondi, V. Chandra, R.R. Bhonde and R. Banerjee, J. Biomater. Tissue Eng., 2, 67 (2012)

NANO-84 Accurate c-myc-Olfactory Receptor quantification on characterized natural nanovesicles for biosensor applications. M. Sanmartí-Espinal1,2, P. Iavicoli1, R. Galve3,4, M.A. Persuy5, E. Pajot-Augy5, M.P. Marco3,4, J. Samitier1,2,4 1IBEC – Institute for Bioengineering of Catalonia, Barcelona, 08028, Spain. 2Department of Electronics, University of Barcelona, Barcelona, 08028, Spain. 3AMRg – Applied Molecular Receptors group, IQAC-CSIC, Barcelona, 08034, Spain. 4Centro de Investigación Biomédica en Red en Bioingeniería Biomateriales y Nanomedicina (CIBER-BBN), 50018, Spain. 5INRA, UR1197 Neurobiologie de l'Olfaction et Modélisation en Imagerie, Jouy-en-Josas, F-78350, France.

Natural vesicles produced from genetically engineered cells with tailored membrane receptor composition are promising building blocks for biosensing devices [1]. This is particularly true for the case of G-protein coupled receptors (GPCRs) present in many sensing processes in cells, whose functionality crucially depends on their lipid environment. However, the controlled production of natural vesicles containing GPCRs and their GPCRs expression level are among the outstanding challenges in the road map to realize practical biomolecular devices based on GPCRs. Largely expressed in the human body, GPCRs are implicated in a wide range of disease pathways [2], including cancer [3,4], cardiomyopathy, and schizophrenia. Around 50% of the drugs on the market regulate GPCR function, 30% of which directly target GPCRs [5]. Herein we present the production and characterization of membrane nanovesicles (nanosomes) of about 100 nm diameter from Saccaromyces Cerevisiae carrying the heterologously expressed olfactory receptor c-myc-OR1740 [6], a member of the family of

GPCRs and representative of important candidates for the development of bioelectronic noses [7]. The nanosomes size in solution was characterized by Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA), and Cryo-Electron Microscopy. In addition, we describe a universal direct novel c-myc-GPCR quantification without the necessity of a previous protein purification step. The quantification was carried out by a highly promising strategy based on a competitive ELISA format assay using c-myc tag-bioconjugates as model conjugates that mimics the behavior of the c-myc olfactory receptor on nanovesicles. A custom made antibody (anti-c-myc) was produced giving assay detectability in the picomolar range and allowing the quantification of

the c-myc-OR proteins. The number of olfactory receptors (OR17-40) per nanovesicle obtained was (30 6) OR/NS. Evidences of the OR17-40 functionality have been obtained by Biacore (SPR) measurements. Present results constitute an important step in the practical realization of biosensor devices based on natural nanovesicles integrating G-protein coupled membrane receptors. [1] V. Sarramegna, F. Talmont, P. Demange and A. Milon Cell. Mol. Life Sci., 60, 1529 (2003). [2] J.S. Gutkind, R.T., Dorsam. Nat. Rev. Cancer., 7, 79, (2007). [3] M. Maggiolini, R. Lappano. Nat. Rev. Drug Discovery., 10, 47, (2011). [4] P.A. Insel, C.M., Tang, I., Hahntow, M.C., Michel. Biochim. Biophys. Acta – Biomembranes., 1768, 4, 994, (2007). [5] E. Jacoby, R. Bouhelal, M. Gerspacher, K. Seuwen. Chem. Med. Chem., 1, 761 (2006). [6] A. Calò, M. Sanmartí-Espinal, P. Iavicoli, M.A. Persuy, E. Pajot, G. Gomila, J. Samitier (submitted to Soft Matter, April 2012) [7] J.M. Vidic, J. Grosclaude, M. A. Persuy, J. Aioun, R. Salesse and E. Pajot-Augy. Lab on a Chip, 6, 1026 (2006).

NANO-85 Biodistribution Study of a Novel Theranostic Versatile Platform Composed of Surface-modified Titanate Nanotubes J. Boudon1, J. Paris1, R. Mayap Talom1, A.-L. Papa1, M. Moreau2, C. Bernhard2, Y. Bernhard2, R. Decréau2, F. Denat2, P. Walker3, A. Oudot4, A. Courteau4, J.-M. Vrigneaud4, B. Collin4 and N. Millot1,*

1 Laboratoire Interdisciplinaire Carnot de Bourgogne, UMR 6303 CNRS/Université de Bourgogne, BP 47870, France 2 Institut de Chimie Moléculaire de l’Université de Bourgogne, UMR 6302 CNRS/Université de Bourgogne, France 3 Service de Spectroscopie par Résonance Magnétique, Centre Hospitalier de Dijon, 2 Boulevard Maréchal de Lattre de Tassigny, 21033 Dijon, France 4 Centre Georges François Leclerc, BP 77980, 21079 Dijon cedex, France

Titanate nanotubes (TiONts) have received more and more attention [1] since their pioneering discovery [2]. Recently, they have been considered for biomedical applications [3] but not yet as a carrier of therapeutic molecules. Previously, our group reported the synthesis of TiONts with a controlled morphology [4] and demonstrated that TiONts were internalized by cells without inducing cytotoxicity [5, 6]. Nevertheless, prior to their use as carriers, it was necessary to determine TiONts biodistribution which has not been reported till date. Our study lies on three biomedical imaging techniques for the non-invasive and real-time in vivo evaluation of these nanovectors: a) SPECT (particularly suitable for such studies [7]), b) MRI with an emphasis on Ultrasmall SuperParamagnetic Iron Oxide (USPIO) particles (widely investigated as T2-MR contrast agent) [8] and c) Optical Imaging (OI). Herein we present the development of a novel carrier based on functionalized TiONts for their use as MRI, SPECT and Optical Imaging (OI) agents. The following pathway was investigated: a direct conjugation was realized to prepare TiONts with functional groups at their surface; a macrocyclic compound chelating agent was covalently attached to the latter functional groups: either 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), for 111Indium labelling (gamma-emitting radionuclide half-life t1/2 ≈ 3 days) or phthalocyanines for optical imaging; a covalent coupling of functionalized PEGs was performed on TiONts conjugates to increase TiONt dispersion, stealth and biocompatibility; USPIO-coated TiONts were also developed to combine both the magnetic detection (MRI) and the delivery of drugs for theranostic applications [9]. The grafting of the organic molecules (DOTA, PEG, phthatlocyanine) and of the USPIO was proven by different analyses and among them

zeta potential, IR, XPS, elemental analysis and thermogravimetric analysis. For the first time, a systematic TiONt-based study is carried out to evaluate the impact of TiONt-conjugates on physiological properties such as blood circulation time and biodistribution in a mouse model. [1] T. Sekino, in Inorganic and Metallic Nanotubular Materials: Recent Technologies and Applications, Vol. 117, p. 17, (Springer-Verlag Berlin, Berlin, 2010). [2] T. Kasuga, M. Hiramatsu et al., Langmuir 14, 3160 (1998). [3] L. Niu, M. Shao et al., J. Mater. Sci. 43, 1510 (2008). [4] A.-L. Papa, N. Millot et al., J. Phys. Chem. C, 113, 12682 (2009). [5] C. Mirjolet, A.-L. Papa et al., Radiother. Oncol. submitted. [6] A.-L. Papa, L. Dumont, et al., Nanotoxicology online. DOI:10.3109/17435390.2012.710661. [7] D. Kryza, J. Taleb et al., Bioconjugate Chem., 22, 1145 (2011). [8] K. M. Krishnan, IEEE Trans. Magn., 46, 2523 (2010). [9] A.-L. Papa, L. Maurizi et al., J. Phys. Chem. C, 115, 19012 (2011).

NANO-86 Multifunctional Hybrid Nano Core@shells via Bioinspired Silification for Targeted Diagnosis

John Wang1 Zhang Yu1, Happy Tan1 and Li Xu2 1Department of Materials Science & Engineering, National University of Singapore, Singapore 117574 2Institute of Materials Research and Engineering (IMRE), Singapore 117602

We have successfully developed a novel bioinspired synthesis strategy, whereby a highly benign approach has been devised to synthesize multifunctional PEOlated Fe3O4 nanocrystals/QDs@SiO2 nano core-shells at room temperature and near-neutral pH condition. The success of such strategy lies on the simultaneous encapsulation of Fe3O4 nanocrystals and QDs into the core of PEO-based polymeric micelles. The encapsulation results in the formation of silica nano-shell being confined to the interface between the core and corona of the Fe3O4 nanocrystals/QDs loaded polymeric micelles. Consequently, the surface of the Fe3O4/QDs@SiO2 nanocapsules is intrinsically covered by a thin layer of free PEO chains, which enable them to be colloidally stable not only at room temperature, but also upon incubation in the presence of proteins under physiological condition. In addition, the silica nano-shell formation does not cause any detrimental effect to the encapsulated Fe3O4 nanocrystals and QDs with respect to their size, morphology, crystallinity, magnetic and fluorescent behavior. The PEOlated nano core-shells have been demonstrated to be an excellent candidate for controlled drug release, magnetic resonance imaging (MRI) and fluorescent imaging contrast agent, as well as a lack of cytotoxicity. In vitro studies of using MDA-MB-231 breast cancer cells showed that the multifunctional nanocapsules were successfully uptaken by the cancer cells and located at the cytoplasm of the cells. Furthermore, by conjugating folic acid on the nanocapsule surface, the uptake by MDA-MB-231 cells is enhanced significantly, suggesting the great potential for targeted imaging and early diagnosis. We will present our latest progress in developing these multifunctional hybrid nano core@shells via bioinspired silification for targeted diagnosis.

NANO-87 Development of coronary artery covered stent using POSS nanocomposite polymer Y. Rafiei1, A. Tan1, A.Darbyshire1, B. G. Cousins1, A. M. Seifalian1,2*

1Centre for Nanotechnology and Regenerative Medicine, Division of Surgery & Interventional Science, University College London, London, United Kingdom 2Royal Free Hampstead NHS Trust Hospital, London, United Kingdom

Endovascular stents have revolutionised the field of interventional cardiology. Despite their excellent clinical outcome complications associated with percutaneous stent implantation following the procedure have remained a major drawback in their widespread use. To overcome such limitations, a number of novel endovascular stents have emerged including a stent wrapped in a thin membrane sleeve. However, due to thethrombogenic characteristics of their covering material the experimental results of these stents were discouraging when verified for restenosis prevention purposesin small diameter vessels. Hence, they are not currently being used as a treatment device for atherosclerosis.In this study we have assessed a nanocomposite polymer comprised of polyhedral oligomericsilsesquioxane (POSS) covalently attached with poly(carbonate-urea)urethane (PCU) as a candidate material for coronary covered stent membranes. This polymer was selected due to its biocompatibility, oxidative resistance, nanotexture and enhanced mechanical properties.POSS-PCU nanocomposite has already been used in the world’s first synthetic trachea, as a lacrimal duct conduit, limb bypass graft and is currently undergoing clinical trial as small diameter coronary artery bypass graft and preclinical trial as a heart valve. A20 µm POSS-PCU membrane was fabricated for coronary artery covered stent. Mechanical testing included compliance, tensile testing, and puncture testing were applied on the polymer. Bonding of polymer to the bare metal surface was assessed using peel strength test and the durability of the polymeric membrane was assessed by exposing the cover stents to accelerated hydrodynamic pulsatile loading for a period of 3 months.Peeling tests and pulsatile flow test showed a good bonding between the polymeric cover and the metal struts.Tensile testing showed POSS-PCU has sufficient ultimate stretch for stent deployment. Puncture testing revealed that POSS-PCU is not likely to puncture in vivo. Percentage of recoil after deployment was similar to that in bare metal stent.Overall, these results indicatethat the use of nanocomposites under optimised conditions could lead to significant advantages in long-term performances and durability of covered stents. NANO-88 Quantum Dot Photosentizer Hybrids as singlet oxygen generators in Photodynamic therapy C. Fowley1, A.P. McHale1, B. McCaughan1, N. Nomikou2 and J.F. Callan1

1Department of Pharmacy and Pharmaceutical Sciences, University of Ulster, Coleraine, Northern Ireland. BT52 1SA 2Sonidel Ltd., Dublin 4, Republic of Ireland.

Photodynamic therapy (PDT) has been used as a clinical treatment since the early 1990s and utilizes a photosensitising drug (PS), molecular oxygen and light of a specific wavelength, (usually visible light below 700nm) to generate singlet oxygen and other reactive oxygen species which are highly cytotoxic [1]. Originally used as a treatment for superficial skin cancer it is now emerging as a treatment for other forms of cancer such as head, neck, lung and prostate cancers. There is, however, several limitations which have prevented PDT obtaining wide spread clinical use. For example currently approved PS drugs absorb light in the visible region limiting tissue depth penetration to a few mm rendering the treatment unsuitable for deep seated tumours. Secondly, PS molecules also tend to be hydrophobic and can aggregate in aqueous solutions, leading to a reduction in singlet oxygen production [2]. Our work seeks to take advantage of the beneficial photophysical properties of Quantum Dots (QDs) to address some of the difficulties currently faced with the use of

conventional PS drugs in PDT. QD-PS conjugates were synthesised allowing the indirect excitation of the PS by energy transfer (FRET) upon excitation of the QD and subsequent production of singlet oxygen. Due to the high two photon absorption capability of QDs these conjugates can be excited with longer wavelength light (> 800nm) which has superior tissue penetration capability. In addition, the QD-PS hybrids displayed greater water solubility than the PS drug alone. Therefore, these conjugates offer promise as new PS agents for use in the treatment of deep seated tumours using PDT. 1. L. M. Davids and B. Kleemann, Cancer Treat. Rev., 2011, 37, 465–475. 2. A. C. S. Samia, S. Dayal and C. Burda, Photochem. Photobiol., 2006, 82, 617–625. 3. W. Zipfel, R. Williams and W. Webb, Nat. Biotechnol., 2003, 21,1368–1376. 4. C Fowley et al, J. Mater. Chem., 2012, 22, 6456-6462.

NANO-89 Polyamidoamine Dextran carrier macromolecules for colon specific delivery of 5-ASA. P.K. Shrivastava1, A. Shrivastava2, S.K. Sinha1, S.K. Shrivastava1

1Department of Pharmaceutics, Indian Institute of Technology, Banaras Hindu University, Varanasi, U.P., India-221 005 2Bansal College of Pharmacy, Anand Nagar, Bhopal, M.P., India- 462021

Aminosalicylates (5-aminosalicylic acid, Mesalamine, 5-ASA) is the drug of choice in the treatment of active inflammatory bowel disease, ulcerative colitis, and Crohn’s disease. It is readily absorbed from the upper gastrointestinal tract (GIT) as soon as it passes through the stomach leading to significant gastrointestinal toxicity. The present study aimed to minimize its gastrointestinal toxicity by sustained and targeted delivery of 5-ASA to the distal ileum and proximal colon and for this dextran (40 kDa) was used as a carrier for targeting 5-ASA at the colonic site using P-Aminobenzoic acid (PABA) and benzoic acid (BA) as linkers. Free carboxyl group of PABA and BA was conjugated with Dextran and further diazotization gives azo polymeric conjugates of 5-ASA. The synthesized and purified conjugates were characterized by UV, HPLC, FTIR and 1H NMR. The degree of substitution was estimated by complete hydrolysis of conjugates in borate buffer, which was found to be 7.5 ± 0.25 %. In-vitro hydrolysis study of conjugates was performed in different biological media, e.g. simulated gastric fluid (SGF) pH 1.2, simulated intestinal fluid (SIF) of pH 7.4 and 3% w/v rat caecal content media of pH 6.8 and the amount of conjugates released was estimated by high performance liquid chromatography. No degradation of conjugates at pH 1.2 and 7.4 showed that the conjugates were stable at the gastric pH. To check the colon specificity of azo polymeric conjugates of 5-ASA, the release pattern in 3% w/v rat caecal content was observed for 24 h and found to be 51.25±1.48 and 47.15±1.03 %. Chronic ulceroginic activity with high ulcer index was obtained for 5-ASA, whereas lower ulcer index was found for the dextran conjugates of 5-ASA.

NANO-90(2) Hemocompatibility of manufactured nanoparticles: Guidelines proposal. J. Laloy1, L. Alpan1, F. Mullier1,2, C. Chatelain2, O. Toussaint3, S. Lucas4, J.M. Dogné1 & B. Masereel1. 1Department of Pharmacy, FUNDP - University of Namur, Namur Nanosafety Center (NNC), Namur Thrombosis and Hemostasis Center (NTHC), NAmur Research Institute for Life Sciences – NARILIS, Namur, Belgium, 2Hematology Laboratory-Namur Thrombosis and Hemostasis Center, CHU Mont-Godinne, Université Catholique de Louvain, Yvoir, Belgium, 3Laboratory of Cellular Biochemistry and Biology, University of Namur, Belgium, 4Research Centre for the Physics of Matter and Radiation, University of Namur, Belgium.

Nanosciences and nanotechnologies are in constant evolution. Development of new therapeutic and diagnostic agents using nanotechnologies for reach their pharmaceutical target require the knowledge of biocompatibility of nanoparticles with the blood compounds. The aim of this study is the evaluation of the biocompatibility of manufactured nanoparticles (NPs) (carbon nanotubes, fullerenes, silicon dioxide, copper oxide and silicon carbide) on erythrocytes integrity, on platelets aggregation and on coagulation cascade. Hemostasis is the ensemble of physiological phenomena which prevent and lead to stop bleeding; it maintains the vascular integrity. A dysfunction of the hemostasis can lead to slow down or even to completely stop the circulation of the blood. It is therefore primordial to know what NPs can have an effect on coagulation. Various techniques assessing activation and aggregation of the platelets or the impact of NPs on coagulation cascade were investigated. An approach in transmission and in scanning electronic microscopy was also accomplished. The Impact-R® with scanning electronic microscopy support and the calibration thrombin generation tests were the reference method to investigate the potential impact of NPs on platelet function and the procoagulant activity of NPs, respectively. Based on the results obtained, we suggest guidelines for testing NP hemocompatibility which responds to a request of scientific community due to lack of recommendations for the evaluation of nanomaterial hemocompatibility. NANO-90 Study of biocompatible and hemocompatible properties of amorphous hydrogenated carbon coatings produced by pulsed magnetron discharge. C. Lopez-Garcia2*, J. Colaux2, J. Laloy1*, F. Mullier1, M. Fransolet3, C. Michiels3, J.M. Dogné1 & S. Lucas2. Namur Nanosafety Centre (NNC), NAmur Research Institute for LIfe Sciences (NARILIS), University of Namur (FUNDP), Rue de Bruxelles 61, B-500 Namur, Belgium. (1) Department of Pharmacy, NAMEDIC, Namur Thrombosis and Hemostasis Center (NTHC), (2) Research Centre for the Physics of Matter and Radiation (PMR-LARN), (3) Research Unit on Cell Biology (URBC).

In this work, we evaluate the potential biomedical application of amorphous hydrogenated carbon (a-C:H) nanofilms deposited by reactive pulsed magnetron sputtering. Our results show a good hemocompatibility and biocompatibility: no effect on hemolysis and hemostasis were observed. Moreover, the proliferation of various cell types such as endothelial, fibroblast and osteoblast-like cells was not affected by the coating. Cell growth on a-C:H coatings is proposed to take place by a two-step process: the initial cell contact is affected by the smooth topography, whereas the polymeric-like structure, a moderate hydrophilicity and a high hydrogen content direct the posterior cell spreading, while preserving the hemocompatible behavior.

NANO-91 Titanium nitride surface layer to optimize vascular endothelial cell growth A.Sowinska1, E. Czarnowska1, T. Borowski2, M. Tarnowski2, B. Walkowiak3, T. Wierzchon2

1The Children’s Memorial Health Institute, Pathology Dept., Warsaw, PL 2 Warsaw University of Technology, Surface Engineering Faculty, Warsaw, PL 3Technical University of Lodz, Department of Biophysics, Lodz, PL

Titanium nitride (TiN) the material accepted by US FDA for medical applications offers particular possibilities for the development of implants with improved haemocompatibility. The TiN coating obtained by PVD methods applied in medicine shows some disadvantages, e.g. easy delamination and limitation of endothelial cells (ECs) biology. Therefore, recent activities focused on improvement of metallic implants haemocompatiblity attempt to alter their properties and direct surface topography to promote among others proliferation of normal ECs. The TiN+Ti2N+αTi(N) surface layer produced under glow discharge conditions by our group has a diffusion character and offers unique combination of high hardness, wear resistance and corrosion resistance followed by wide range of biocompatibility. Thus, the aim of our approach was to modify outer zone of nitrided surface layer to create favourable surface nanostructure to enhance ECs development and decrease platelets adhesion. Two different low temperature (680oC) processes of glow discharge were applied to manufacture TiN+Ti2N+αTi(N) surface layers on titanium alloy Ti6Al4V base using the cathode or plasma region (so called active screen) nitriding process. The outer surface of the layers marked as TiN-K and TiN-P exhibited thickness about 1,0 and 0,5 μm, respectively. Both surfaces exhibited fine grain structure visualized under AFM. The mean roughness parameters of TiN-K and TiN-P measured under AFM were Ra= 67.09 nm and 9.44 nm and Rq= 82.82 nm and 13.00 nm, respectively. Surface wettability investigated by water contact measurement was comparable for TiN-P and TiN-K surface. Biocompatibility in vitro was investigated with HUVEC endothelial cells in range of adhesion, proliferation, cell cycle under laser scanning cytometer and confocal microscope and with platelets (from human PRP samples) in range of adhesion and aggregation under SEM. The study revealed significantly better interaction and activity of ECs and similar platelet activation and higher adhesion on TiN-P comparing to TiN-K. Significance of the surface rougness combined with nanostruture and others surface layer properties for biocompatiblity is discussed. This study was supported by the National Science Centre [grant 2011/01/D/ST8/02931]. NANO-92 Nanoparticles of Alkylglyceryl-Dextran-poly(lactic acid) for Drug Delivery to The Brain P. Toman1, C.F. Lien1, D. Górecki1, J. Tsibouklis1, G. Pilkington1, J.R. Smith1, Z. Ahmad1, Q. An1, E. Barbu1

1School of Pharmacy and Biomedical Sciences, University of Portsmouth, UK

Of the several non-toxic, biocompatible and biodegradable polymers that are available, poly(lactic acid) – a material known for its inherent tendency to form colloidal systems of low polydispersity – and alkylglycerol-modified dextran – a material designed to combine the drug carrier properties of dextran with the permeation enhancing properties of alkylglycerols – have been combined for the development of nanoparticulate, “blood brain barrier-permeating” vectors. To this end, dextran that had been functionalised via treatment with epoxide precursors of alkylglycerol was covalently linked to poly(lactic acid), using carbodiimide, to form alkylglyceryl-modified dextran-poly(lactic acid). The Solvent Displacement Method allowed the formation of this, well-defined (as confirmed by 1H-, 13C-NMR and FT-IR) and highly purified (dialysis), product into nanoparticles with a narrow size distribution profile. The relative uniformity of size in the solid state was confirmed by the Scanning Electron Microscopy and Atomic Force Microscopy images of freeze dried samples, while that of nanoparticles in liquid

media was confirmed by Dynamic Light Scattering, Nanoparticle Tracking Analysis and Electrophoretic Mobility Measurements on freeze-dried nanoparticles that had been re-dispersed in phosphate buffer (pH 7.4). The drug-loading capacity of nanoparticles has been modelled by preparing nanoparticulate congeners incorporating the easy-to-detect dye Rhodamine B. The capability of nanoformulations to open the tight junctions has been suggested by Transwell-model experiments employing mouse- (bEnd3) or human- (hCMEC/D3) brain endothelial cells and also by Electric Cell Substrate Impedance Sensing. In parallel, the biocompatibility of nanoparticles was indicated by the MTT and Presto Blue assays, while cellular uptake was evidenced by confocal microscopy. The collective in vitro data suggest that these alkylglyceryl-modified dextran-poly(lactic acid) nanoparticles are promising candidates for in vivo evaluations that would test their capacity to transport therapeutic actives to the brain. NANO-93 Short biocompatible nanofibres as a flexible building block for nano-scaffolds in tissue engineering A. Sutti1, C. Wong1, R. Rajkhova1, B. Allardyce1, A. Gestos1, X. Wang1

1Institute for Frontier Materials, Deakin University, Waurn Ponds, VIC 3216, Australia.

Short nanofibres were prepared from a selection of natural and synthetic polymers, using a newly developed solution dispersion technique, and were used to produce a series of nanostructured scaffolds suitable for cell growth. Short nanofibres are fibres which diameter is sub-micron, with aspect ratio above 50.1, 2 Short nanofibres present numerous advantages in comparison with electrospun nanofibres, though short nanofibres are yet to be widely employed, due to their very recent appearance.1 The main advantage in using short nanofibres3 is the ability to employ the fibres as such, for instance to prepare compact nanofibre mats, as well as to use the fibres as one would nanoparticles, as in the preparation of monolayers or freeze-dried open extracellular matrix mimics. Such peculiar dual nature endows short nanofibres with great potential in building nanostructures at different scales. Great flexibility in scaffold design was achieved in this work thanks to this dual nature of short nanofibres. 2D as well as 3D scaffolds were produced for cell culture, some of which presenting hierarchically roughness. Examples of the versatility of this new type of material, short nanofibres, will be presented, with the related cell culture data to demonstrate their potential in scaffolds for tissue engineering. The relation between scaffold design and cell growth will be presented and discussed. 1. A. Sutti, T. Lin and X. Wang, Journal of Nanoscience and Nanotechnology 11, 1-6 (2011). 2. R. G. Alargova, V. N. Paunov and O. D. Velev, Langmuir 22 (2), 765-774 (2005). 3. F. Balzer, J. Ihlemann, A. C. Simonsen and H.-G. Rubahn, Proc. SPIE 5720, Micromachining Technology for Micro-Optics and Nano-Optics III,165-172

(2005).

NANO-94 Morphology Characterization and In-Situ Small Angle X-ray Scattering Study of Materials Transport through Hollow Nanoparticles for Drug Delivery Z. H. Chen1, X. B. Zeng1, G. Ungar1, 2, S. H. Hwang2, C. Kim2, J. Jang2, G. Newby3

1Department of Materials Sci. & Eng., University of Sheffield, Sheffield S1 3JD, U.K. 2WCU program of Chemical Convergence for Energy & Environment, School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, Korea 3Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, U.K.

SiO2/TiO2 spherical hollow nanoparticles (HNPs) were synthesized by using silica nanoparticles as templates [1]. Morphological parameters of HNPs, i.e. average outer diameter, inner diameter, polydispersity, porosity of the shell and size of intrawall pores, were extracted by a combination of experimental methods, including transmission and grazing incidence small angle X-ray scattering (SAXS and GISAXS), small angle neutron scattering (SANS), transmission electron microscopy (TEM), dynamic light scattering (DLS), and N2 adsorption-desorption isotherms [2]. The porosity of the shell of HNPs was found to be as high as 40%. In order to test the models used in deriving porosity and surface area from vapour-pressure / volume isotherms, deposition of liquid layers on the surfaces and condensation within the central void and intrawall pores was monitored by SAXS using a specially constructed in-situ humidity cell. HNPs with different surface functionalities were studied. Furthermore, to investigate material transport through HNP walls during drug delivery, filling and emptying of HNPs by solvents of different electron densities and hydrophilicities were monitored directly by in-situ synchrotron based SAXS. The time-dependent scattering function was modeled by a 3-level electron density profile in which the level in the centre of the particle was allowed to change with time. The above study suggests that SAXS may become a valuable in-situ method for monitoring material transport in drug delivery vectors. [1] M. Choi, C. Kim, S.O. Jeon, K.S. Yook, J.Y. Lee and J. Jang, Chem. Commun., 47, 7092 (2011). [2] Z.H. Chen, C. Kim, X.B. Zeng, S.H. Hwang, J. Jang and G. Ungar, submitted to Langmuir.

NANO-95 Biological Evaluation of Carbon Layers Doped With Silver W. Jakubowski1, P. Komorowski1, D. Bociaga2, D. Batory2, P. Niedzielski2, B. Walkowiak1

1Department of Biophysics, Technical University of Lodz, Poland 2Department of Biomedical Engineering, Technical University of Lodz, Poland

Progress in biological sciences and medicine is today increasingly dependent on collecting and examining information on the relationship between the cells of living organisms and abiotic surfaces of different materials. This is especially important in areas related to the use of biomaterials. Interactions between material and living cells have until recently been largely considered outside the scope of the standard use of biomaterials. In the process of developing materials with a high degree of biocompatibility, much of the research has focused on the problem of abiotic surface colonization by different microorganisms. One of the most significant challenges in developing new biomaterials is the reduction or elimination of biomaterial surface colonization, which otherwise usually leads to the development of microbial biofilm. One possible modification of material surface, which could reduce microorganisms colonization, is an introduction of elements that exhibit bactericidal and fungicidal activity. The Institute of Materials Science and Engineering has developed technology of production using a hybrid method RF PACVD/MS, which combines the bioinert nature of carbon layers and the bactericidal properties of silver ions. Such a coating can significantly reduce the degree of bacterial adhesion and results in limitation of the development of a biofilm on the materials surface, whilst having a negligible cytotoxic effect on the cells of higher

organisms. Results of experiments conducted with the model bacterium E. coli (DH5α) culture together with human osteoblasts (Saos 2) and endothelial cells (EA.hy 926) fully confirmed the assumed properties of this new material. This research was financed by project ERA -NET/MNT/CARSILA/1/2010. NANO-96 AFM Measurement of Single Cell Mechanical Properties of Live, Membrane-Impaired and Dead Human Mesenchymal Stem Cells

Nikolay I. Nikolaev1, Torsten Müller2, David J. Williams1 and Yang Liu1

1Wolfson School of Mechanical & Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK; 2JPK Instruments Limited, Unit 7223, Cambridge Research Park, Waterbeach, Cambridge CB25 9TL, United Kingdom In our previous research1, it has been found that human mesenchymal stem cells (hMSCs) were only sensitive to certain vibration conditions and there was a synergistic effect of cold storage duration and vibration-induced mechanical cell damage. As it is well known that prolonged cold storage induce apoptotic and necrotic cell death, in the current study AFM nano-indentation was carried out to identify the mechanical properties of single suspended viable, dead and membrane-impaired hMSC after cold storage. iable hMSC have Young’s modulus in the range of 0.7~1.1kPa and consistent measurement was observed even when different measurement site was chosen. Among the hMSC cells analysed, different intensity of Annexin V and PI staining were observed, showing cells with different extent of membrane damage. The mechanical properties of cells with impaired membrane as indicated by Annexin + and PI- depended on the location of the measurement. The measurement of Young’s modules of cell with some membrane impairment as indicated by a few phospholipid phosphatidylserines presented on cell surface was 0.943 ± 0.640 kPa or 4.528 ± 2.094 kPa , depending on the site measured. In summary, the mechanical properties of cell membrane could be applied as a indictor of cell healthy state and further research on different type of cells is needed to confirm this finding in order to allow categorisation and separation of cell populations by correlating the cell physical properties with its biological function. [1] N. I. Nikolaev, Y. Liu, H. Hussein and D. J. Williams, J. R. Soc. Interface 7 October 2012 vol. 9 no. 75 2503-2515

NANO-97 Attenuation of glutathione synthesis enhances disulfonated meso-tetraphenylporphyrin (TPPS2a) – mediated nano-delivery of saporin in breast cancer cells. D K Adigbli, J Seebaluck, M Loizidou, A J MacRobert National Medical Laser Centre, Div. of Surgery and Interventional Science, UCL Medical School, UCL, London, UK.

Photochemical internalisation (PCI) is a light- and oxygen- dependent technique that facilitates nano-delivery of molecules to their intracellular targets. Molecules, including chemotherapeutics, are internalised into endocytic vesicles that may subsequently fuse with lysosomes for eventual degradation. PCI utilises amphiphilic photosensitisers (PS), which localise in the endo/lysosomal membranes. In the presence of light and oxygen, PS produce reactive oxygen species (ROS) that cause localised damage, at a nono-scale, to the vesicle membrane resulting in the release of the molecules into the cytoplasm. In doing so PCI potentiates the endocytic-dependent delivery of molecules inside the cell. Buthionine sulfoximine (BSO) is a synthetic amino acid that irreversibly inhibits gamma-glutamylcysteine synthetase, the enzyme required in the first step of glutathione synthesis. In doing so BSO attenuates intracellular levels of the powerful reducing agent glutathione, thereby increasing

susceptibility to oxidative damage. It is already used clinically to help reverse glutathione-mediated resistance to alkylating agents and platinum compounds in cancer chemotherapy. To determine whether BSO can potentiate disulfonated meso-tetraphenylporphyrin (TPPS2a) – mediated PCI of the type I ribosome inactivating protein Saporin (SAP) in breast cancer cells….f instead of ph is standard international usage now. 4T1 (murine breast cancer) cells were incubated with SAP, BSO and TPPS2a separately or in various combinations. Cells were then exposed to blue light (420nm) or kept in the dark throughout (controls). Viability was determined using the MTT assay. Using dose response baseline experiments, the chosen dose of TPPS2a alone (0.6µg/ml) demonstrated 37% killing (PDT effect, p<0.01) following exposure to blue light for 150 seconds. SAP alone (40nM) exerted a minimal (<3%) kill effect. BSO alone (0.5µg/ml and 1.0µg/ml) killed <4% of cells. In combination the two drugs (TPPS2a + SAP) killed 60% of cells (p<0.01), consistent with PCI. This represents a 23% and 57% (p<0.01) increase in cytotoxicity compared to TPPS2a or SAP alone, respectively. Addition of BSO resulted in a 13% (BSO 0.8µg/ml, p<0.05) and 25% (BSO 1.0µg/ml, p<0.01) increase in cell killing. We found this effect was independent of any increase in TPPS2a plus BSO cytotoxicity (PDT effect). Our study also observed that by splitting the total illumination time into two divided doses over a 4-hour period, we were able to attenuate the PDT effect, whilst enhancing PCI. By enhancing the kill effect of TPPS2a in combination with SAP, BSO supports the role of reactive oxygen species in mediating the release of endocytosed molecules from endo-lysosomes in PCI. Furthermore, given its previous use in cancer chemotherapy, BSO may prove a useful adjunct to PCI-mediated drug delivery in cancer. NANO-98 Electrospun Nanofiber Mediated Stimuli Responsive Drug Release M. Chen,1 F. Besenbacher1

1Interdisciplinary Nanoscience Center (iNANO), University of Aarhus, Aarhus, Gustav Wieds Vej 14, DK-8000 Denmark.

Drug delivery development faces numerous challenges, such as improved efficacy and specificity, reduced side effects, sustained release, reduced pain from administration, increased ease of use and safety etc. New classes of pharmaceuticals and biologics (peptides, proteins and DNA-based therapeutics) that are fueling the rapid evolution of drug delivery technology cannot be effectively delivered by conventional means. Rather than through conventional burst release techniques (such as oral ingestion, injection), novel delivery techniques with controlled release are on demand. Consequently, delivery vehicles and drugs become more tightly coupled in the delivery design.[1] Advances in nanotechnology and biotechnology will create new opportunities to develop biomimetic smart materials to meet the design requirements. Electrospinning, one of the most exciting nanotechnology innovations in recent years, is a robust, straightforward, and powerful method to produce nanofibers[2] with the potential to incorporate drugs in a simple, rapid, and reproducible process. The review summarizes ongoing researches that utilize electrospinning, one of the top-down nanotechniques, to fabricate nanofibrous form of current researched bottom-up intelligent materials as delivery vehicles, e.g., that are responsive to certain key environmental factors, such as temperature, light and pH, to allow stimuli-responsive control release of the bio-actives in a timely and spatially controlled manner. Furthermore, the electrospun nanofibers that mimic extracelluar matrix intrinsically guide cellular drug uptake, which will be highly desired to translate the promise of drug delivery for the clinical success. 1. R. Langer, and D.A. Tirrell, Nature, 2004. 428 (6982): p. 487-492. 2. Luo, C.J., et al., Chemical Society Reviews, 2012. 41(13): p. 4708-4735.

NANO-99 Electrospun Scaffolds for Annulus Fibrosus Tissue Engineering R. Kang,1 M. Chen,2 C. Bünger,1 F. Besenbacher2

1 Orthopaedic Research Lab, Aarhus University Hospital, Noerrebrogade 44, DK-8000 Aarhus, Denmark. 2 Interdisciplinary Nanoscience Center (iNANO), University of Aarhus, Aarhus, Gustav Wieds Vej 14, DK-8000 Denmark.

Damage and/or degeneration of the disc is a common occurrence, afflicting upward of 97% of the population by 50 years of age. During degeneration, the annulus fibrosus (AF) becomes progressively disorganized, concomitant with mechanical and structural failure including tears, fissures, and delamination, each of which is thought to contribute to low back pain.[1] Treatments for discogenic back pain and disc degeneration are largely palliative, and restoration of function remains unaddressed. Therefore, there is great need for regenerative strategies that may alleviate low back pain while restoring mechanical function. Electrospinning permits the fabrication of highly aligned arrays of polymeric nanofibers, whose scale and architecture can be manufactured to possess key mechanical behaviors of fiber-reinforced soft tissues including nonlinearity, anisotropy, and finite elastic deformations.[2] Moreover, fiber alignment in the scaffold guides alignment of resident cells and ordered deposition of extracellular matrix (ECM). In this study, we reinforced electrospun nanofibrous scaffolds with fused deposition modeling (FDM) macrofibers to develop a multilamellar engineered tissue that replicates the multi-scale architecture of the native AF. Attachment and proliferation on electrospun scaffolds have been observed for porcine AF cells and human mesenchymal stem cells (MSCs). [1]. An, H.S., E.J.M.A. Thonar, and K. Masuda, Spine, 2003. 28(15): p. S86-S92. [2]. Agarwal, S., J.H. Wendorff, and A. Greiner, Advanced Materials, 2009. 21(32-33): p. 3343-3351.

NANO-100 Polymersomes as nanocarriers for radionuclide imaging G. Wang1, R. de Kruijff1, E. Mendes2, H. T. Wolterbeek1, A. G. Denkova1

1Radiation, Radionuclides & Reactors, Delft University of Technology, Mekelweg 15, 2629 JB Delft, The Netherlands 2Department of Chemical Engineering, Delft University of Technology, Julianalaan 136, 2628 BL Delft, The Netherlands

Polymersomes are polymer vesicles which are self-assembled structures composed of amphiphilic block copolymers that form a hydrophobic bilayer enclosing an aqueous cavity. Although polymersomes have recently shown to have great potential for use in medical applications [1], studies that account for their use in nuclear medicine are strikingly low [2]. Here, we demonstrate that polymersomes composed of poly(butadiene-b-ethylene oxide) (PB-PEO) can be loaded with radionuclides (111In and 64Cu) through an active transport mechanism that uses a lipophilic ligand such as tropolone to transport the radionuclide through the hydrophobic membrane. Loading efficiency above 80 % are achieved for both radionuclides allowing the encapsulation of sufficient amount of activity to allow their use in nuclear imaging techniques such as Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET). In addition loss of radiolabel has been found to be less than 1.5 % ensuring safe application. 1. J. Z. Du and R. K. O’Reilly, Soft Matter. 5, 3544 (2009) 2. R. P. Brinkhuis, K. Stojanov, P. Laverman, J. Eilander, I. S. Zuhorn, F. P. J. T. Rutjes and J. C. M. van Hest, Bioconjugate Chem. 23, 958 (2012)

NANO-101 Evaluation of carbon nanocoatings deposited onto polymer parts of artificial heart

1W. Kaczorowski, 1D. Batory, 1W. Jakubowski, 1W. Okrój, 1M. Walkowiak-Przybyło, 2A. Szuber, 2R. Kustosz, 1S. Mitura, 1P. Niedzielski

1Lodz University of Technology, Lodz, 1/15 Stefanowskiego St. Poland 2 Foundation for Cardiac Surgery Development, Zabrze, 345 Wolności St. Poland

Surface modifications of biomaterials with use of carbon coatings are described in the literature as one of the most popular for the improvement of their biocompatibility. Due to the possibility of control of their mechanical properties (hardness, modulus) they are very promising material for modification of variety of medical implants made of metals, alloys and polymers as well. In this work are presented results of modification of polymer substrates used in artificial heart construction (polyurethane PU and PEEK) by means of the synthesis of carbon nanocoatings and nanocomposite carbon films doped with titanium and silicone. Thickness of obtained layers was in the range between 50 – 180 nm. They contained high amount of sp2 bonded carbon (up 85%), nevertheless they increased the hardness of the substrate surface up to five times (1,6 GPa) and modulus up to three times (14,6 GPa). In presented work are reported physicochemical properties of carbon based nanocoatings obtained by means of AFM, XPS, Raman spectroscopy and nanoindentation methods. Evaluation of the biological properties of synthesized coatings was performed on the basis of the analysis of thrombogenicity examined with use of the Impact-R method by the evaluation of the level of platelets activation and adhesion of particular blood cell elements. Additionally the analysis of antimicrobial resistance of E. Coli colonization was performed. Obtained results prove that polymer materials (PU, PEEK) modified with use of carbon nanolayers can constitute a very interesting solution for the construction of artificial hart. Applied carbon coatings show the protective activity but do not generate increased blood activation. The works have been financially supported within "Development of the type series of single disk artificial heart valves for paediatric ventricular assist devices" research project (NR13-0118-10/2011) by The National Centre for Research and Development. NANO-102 DEVELOPMENT AND OPTIMIZATION OF POLYMERIC NANOPARTICLES OF ANTITUBERCULAR DRUGS USING CENTRAL COMPOSITE FACTORIAL DESIGN R.Chawla, S. Jaiswal, B. Mishra Department of Pharmaceutics, Indian Institute of Technology (Banaras Hindu University) Varanasi-221005, India

The objective of the present study was to statistically design, optimize and develop polymeric nanoparticles (PNs) for two Anti-tubercular drugs Isoniazid and Rifampicin using Resomer RG 752 S (PLGA) and Poloxamer 188 by double emulsification evaporation technique. A two-factor, five-level, 22 + star circumscribed central composite design was employed to study the influence of critically selected formulation factors (independent variables), surfactant concentration and polymer amount, on the physicochemical characteristics (dependent variables) such as particle size (PS), polydispersity index (PI) and entrapment efficiency (EE) of the PNs. Morphological characterization, physicochemical evaluation and stability studies were also performed as a part of optimization studies. The results of the study suggested that using PLGA twice the amount of drug and poloxamer concentration upto 2.0 % w/v resulted in efficient EE for both drugs [1]. Using the response surface regression (composite desirability of 0.899) the optimized PNs were prepared which exhibited spherical and porous surface with a mean PS of 202 nm, PI of 0.178 + 0.015, zeta potential of -25.49 mV and EE of 76.12% and 54.25 % for RIF and INH respectively. The in vitro studies performed at physiological pH 7.4 and endosomal macrophage pH 5.2 by dialysis bag diffusion technique showed sustained release of drugs up to 48 hrs. In vivo

qualitative uptake studies of Rhodamine labeled PNs in peritoneal macrophages assessed through fluorescence microscopy and Confocal Laser Scanning Microscopy suggested that the PNs were endocytosed sufficiently without showing any surface adsorption phenomenon [2]. These results could be further exploited for the development of a suitable targeted delivery system for the treatment of Tuberculosis. [1] R. Pandey, Z. Ahmed, S. Sharma and G.K. Khuller, Tuberculosis, 83, 373 (2003) [2] V Kanchan and A. K. Panda, Biomaterials 28, 5344 (2007)

NANO-103 Rare-earth ion conjugated metabolites for imaging biomarkers for cardiovascular disease S. Sahaa, G. Joseb, T. Kakkarb, D.P. Jatib, J. Coxa, C. Bauera, J. Boylea, C. Peersa, A. Jhab

a Leeds Institute of Genetics, Health and Therapeutics (LIGHT), Faculty of Medicine and Health , University of Leeds, United Kingdom. b Institute of Materials Research, School of Process, Environmental and Materials Engineering, University of Leeds, United Kingdom.

Cardiovascular disease (CVD), which includes ischemic stroke and acute myocardial infarction, is the leading cause of global mortality and represents major economic burden on health care systems. Ischemic events in the brain and cardiovascular system result in accumulation of lactic acid and other metabolic biomarkers [1]. Here we have investigated the possibility of using lanthanides [europium (Eu3+), ytterbium (Yb3+) and cerium (Ce3+/4+)] as potential conjugates for metabolite analysis. We have also tested cyto-toxicity of these ions and found that cell viability of neuroblastoma (SH-SY5Y) increased (as determined by MTT assays) when treated with these ions

(1- 1000 M). The cyto-protective influence was also observed with Ce3+/4+ and Eu3+, when the cell lines were exposed to oxidative stress. Characterization and profiling of these lanthanides’ excitation and bond vibration signature have been successfully performed, both individually and when combined with various samples containing metabolites (e.g., lactic acid, phospholipids). Our results show a unique dependence of visible photoluminescence in Eu3+ resulting from 5Dm to 7Fn transitions, which can be exploited for early detection of metabolites for CVD through non-invasive approaches. We have also photo-tagged Yb3+ and Ce3+/4+ with lactic acid and phospholipids and studied their spectroscopic properties. Yb3+ shows more chemical environment sensitivity than Ce3+/4+. The uniqueness of the Eu3+ photoluminescence intensity ratio, as an example, reported here for metabolite imaging has potential for integration of the conjugated molecular techniques with the standard near-IR device technology which may be useful in early detection and diagnosis of a number of critical diseases. [1] S.H. Shah, W.E. Kraus and C.B. Newgard, Circulation. 126:1110 (2012)

NANO-104 Multifunctional squalenoyl nanocomposites for pancreatic cancer: diagnostic and therapeutic aspects S. Valetti1,2, S. Mura1, B. Stella2, D. Desmaële1, M. Noiray1, F. Maione3, E. Giraudo3, L. Cattel2, P. Couvreur1. 1UMR CNRS 8612, Institut Galien Paris-Sud, Université Paris-Sud, 92296 Châtenay-Malabry, France 2Dipartimento di Scienza e Tecnologia del Farmaco, Università degli Studi di Torino, 10125 Torino, Italy 3Institute for Cancer Research and Treatment, 10060 Candiolo, Torino, Italy

The “squalenoylation” nanotechnology consists in the linkage of squalene, a natural and well tolerated triterpene, to anticancer drugs and antiviral nucleoside analougues. This stategy, which allows to achieve a high drug loading together with a high therapeutic efficiency and the possibility to overcome the resistance to current available treatments [1], represents a new and original approach for delivering therapeutic compounds. The anticancer

activity of Squalenoyl Gemcitabine (SqGem) nanoparticles has been already investigated both in vitro and in vivo showing their highest efficacy compared to the free drug both on several tumor cell lines and on leukemic mice [2]. Furthermore, these nanocarriers result as a good platform to develop a theranostic approach for personalized therapy [3]. The aim of this project is to design a novel theranostic multifunctional nanomedicine which combines multi-therapy, tumor targeting as well as imaging functionalities. Nanoparticles for multi-therapy (anti-angiogenic and chemotherapy) were prepared by the self-assemblage in water of two squalene-based prodrugs (SqGemcitabine and SqSunitinib). Their surface was decorated with a specific ligand (i.e. CKAAKN peptide) able to selectively target pancreatic cancer cells [4]. Imaging properties were obtained via the encapsulation of iron oxide nanoparticles (USPIO) into these multifunctional nanoparticles. Physico-chemical characterization and stability studies of this nanoparticle were performed. Preliminary results (Surface Plasmon Resonance analysis and in vivo tests) make these multifunctional nanoparticles extremely appealing for application in pancreatic cancer management. [1] D. Desmaële, R. Gref, P. Couvreur, Squalenoylation: A generic platform for nanoparticular drug delivery. Journal of Controlled

Release doi:10.1016/j.jconrel.2011.07.038 (2011). [2] L.H. Reddy, P.E. Marque, C. Dubernet, S.L. Mouelhi, D. Desmaele, P. Couvreur, Preclinical toxicology (subacute and acute) and

efficacy of a new squalenoyl gemcitabine anticancer nanomedicine. J Pharmacol Exp Ther, 325, 484 (2008). [3] J.L. Arias, L.H. Reddy, M. Othman, B. Gillet, D. Desmaele, F. Zouhiri, F. Dosio, R. Gref, P. Couvreur, Squalene Based Nanocomposites:

A New Platform for the Design of Multifunctional Pharmaceutical Theragnostics. Acs Nano, 5, 1513 (2011). [4] J.A. Joyce, P. Laakkonen, M. Bernasconi, G. Bergers, E. Ruoslahti, D. Hanahan, Stage-specific vascular markers revealed by phage

display in a mouse model of pancreatic islet tumorigenesis. Cancer Cell, 4, 393 (2003).

NANO-106 Generic graphene biosensor for detection of disease biomarkers Owen J Guy, Z. Tehrani, G. Burwell, S. Teixeira, M.A. Mohd-Azmi, and A. Castaing Swansea University, Singleton Park, Swansea SA2 8PP, UK.

Graphene devices and sensors promise to be a disruptive technology in next generation electronics and healthcare diagnostic applications - due to graphene’s superlative properties. Epitaxial graphene is the most amenable form of graphene to electronics, as devices can be fabricated using standard semiconductor wafer processing techniques [1]. Chemical functionalisation of graphene can be used to tailor the chemical specificity and electronic properties of graphene via doping and band-gap engineering [2]. Graphene is therefore an ideal material for fabrication of electrochemical biosensors [3]. Generic biosensor technology, based on functionalized epitaxial graphene (grown on silicon carbide), has been developed. The electrochemical functionalisation technology reported provides a generic platform for the attachment of any number of antibodies or other bioreceptor molecules. Several antibodies including those targeted against Beta-actin, 8-OHdG, pregnancy marker hCG, and cardiac markers Troponin and Myoglobin, have been covalently attached to graphene. By covalently attaching monoclonal antibodies to arrays of semiconductor devices, addressable nano-channels can be fabricated that demonstrate a selective current response to the biomarker analyte. Surface characterization using XPS, Raman spectroscopy, and fluorescence microscopy has been used to confirm that monoclonal antibodies can be selectively attached to graphene channels, functionalized under an electrical bias. Electrical characterization of channels demonstrate that the graphene sensors enable highly sensitive (nM) detection of disease biomarkers. [1] A. Tzalenchuk, S. Lara-Avila, A. Kalaboukhov, S. Paolillo, M. Syväjärvi, R. Yakimova, O. Kazakova, T. Janssen, V. Fal'Ko, S. Kubatkin, Nature nanotechnology 5, 186 (2010). [2] J.M Englert, C. Dotzer, G. Yang, M. Schmid, C. Papp, J.M. Gottfried, H.-P. Steinrueck, E. Spiecker, F. Hauke, A. Hirsch, A. Nature Chemistry 3, 279 (2011). [3] O.J. Guy, G. Burwell, Z. Tehrani, A. Castaing, K-A. Walker, K, S.H. Doak, Materials Science Forum 711, 246 (2012).

NANO-107 Integrated Silicon Nanowire Biosensor Devices for Detection of Cancer Risk Biomarker Owen J Guy, M.A. Mohd-Azmi, Z. Tehrani, R.P. Lewis, K-A. D. Walker Swansea University, Singleton Park, Swansea SA2 8PP, UK.

Bio-functionalisation of SiNW channels with “bioreceptor” molecules capable of specific and selective binding with disease biomarkers is an important enabling technology in the development of nanoscale diagnostic sensors for detection of disease biomarkers using highly sensitive, label-free, electrochemical, SiNW chips. A novel sensor device, based on a silicon nanowire (SiNW) array fabricated on silicon-on-insulator (SOI) substrates, is presented. The SiNW sensor consists of a SiNW functionalised with a “bioreceptor” antibody which can interact selectively with target biomarker. By attaching aniline to SiNW, via coupling with an aryl diazonium salt, the amino group of the aniline molecule has been used to graft antibodies - (1) targeted against beta-actin and (2) targeted against 8-hydroxydeoxyguanosine (8-OHdG) - onto the SiNW surface. Antibody attachment to SiNW surfaces has been verified using fluorescence microscopy to detect quantum-dot labelled antibodies bound to the SiNW surface. Device packaging, electrical analysis and display systems have also been developed for the SiNW sensor. SiNW devices bonded on a “bio-smartcard”, fabricated using printed circuit board technology, are subsequently slotted into a “card-reader” readout device. SiNW sensors are susceptible to significant changes to their electronic transport properties in response to changes that occur at their surfaces. The biomolecular interaction at the SiNW surface can thus be detected, via a current-voltage measurement, using the Point-of-Care (POC) bio-smartcard and card reader in readout mode. The silicon nanowire devices show an increase in the resistivity of the SiNW upon chemical functionalization with aniline and further increases in resistivity on attachment of the bioreceptor antibody and subsequent binding with the disease biomarker. SiNW sensors have been used to detect the presence of target biomarkers at nM concentrations. [1] A. Gao, N. Lu, P. Dai, T. Li, H. Pei, X. Gao, Y. Gong, Y. Wang, C. Fan Nano Letters, 11, 3974 (2011).

NANO-108 Study of Melanoma Cells Apoptosis Induced by Magnetic Hyperthermia A. Avram1, 2, M. Volmer3, 1, V. Schiopu1, I. Petrescu4, M. Avram1

1 National Institute for Research and Development in Microtechnologies, Bucharest, Romania

2 Politehnica University of Bucharest, 313 Splaiul Independentei, Bucharest, Romania

3 University “Transilvania” Brasov, Romania

4 Department of Plastic Surgery and Reconstructive Microsurgery, Emergency University Hospital, Bucharest, Romania

This study examines the apoptosis of B16 mouse melanoma subjected to magnetic hyperthermia. Maghemite superparamagnetic nanoparticles, synthesized and characterized by SEM, X-ray diffraction, FTIR spectrometry and VSM, were successfully tested in the treatment of B16 melanoma induced artificially to lab mice [1]. Therapeutic protocol consisted of two sessions of intravenous nanoparticle administration, followed by a local exposure to a magnetic field concentrator and to an external alternating magnetic field (120 kHz and 18 mT) for 30 minutes. The investigations have shown that an aqueous suspension of polyethylene glycol coated maghemite, exhibits excellent magnetic properties under the influence of AC magnetic fields [2]. Their specific loss power may be well understood on the base of supra-position of Néel and Brown relaxation processes taking into account the actual particle size distribution [3]. FTIR characterization revealed cellular apoptosis after magnetic hyperthermia treatment was applied to B16 melanoma mice. The appoptosis appears due to the occurrence of the oxidative process with fatty acids in the tumour tissues. The spectral band at 1283 cm-1, band that can be

assigned to the phosphatidylserine bond, confirmes the apoptosis of the melanoma cells treated through magnetic hyperthermia. The translocation of phosphatidylserine from the inner to the outer leaflet of the plasma membrane is an early event in apoptosis. [1] M. Yonezawa, et al., Int. Journal of Cancer, 66(3), pp.347-351, 1996. [2] S. Balivada, et al., BMC Cancer, 10:119, 2010. [3] . aradan, L. Chen, J. Xie, “Nanomedicine Design & Applications of Magnetic Nanomaterials, Nanosensors, Nanosystems”, John Wiley & Sons, pp.55,152-175, 2008.

NANO-109 Quantum Dots Surface Modification with Bisphosphonates S.F. Abdul Ghani1, E. Rosca1, M. Wight1, N.J. Long2, M. Thanou1

1Institute of Pharmaceutical Science, School of Biomedical Sciences, King’s College London 2Department of Chemistry, Faculty of Natural Sciences, Imperial College London

In this project we prepare quantum dots based on core cadmium selenium, with zinc sulphide as the shell (CdSe/ZnS). These quantum dots are designed to be used in pharmacokinetics studies in tumours using in vivo imaging. Quantum dots CdSe/ZnS offer excellent fluorescent properties with narrow size distribution and tuneable emission spectrum that are suitable for bioimaging. However, these quantum dots are required to go through surface modifications in order to improve aqueous dispersion and biocompatibility. CdSe/ZnS_TOPO were synthesised following the method of Bawendi et al with slight modifications. Surface modification has been carried out through ligand exchange reaction in which TOPO layer was replaced by bisphosphonates (BIPs) such as ethylenediphosphonic acid (EDP), methylenediphosphonic acid (MDP) and imidodiphosphate sodium salt (IDP). Purification protocols have been developed using centrifugation, PD-10 desalting column and mini dialysis (MWCO 10K). Transmission electron microscopy (TEM) and fluorescent spectroscopy have been used to characterise quantum dots and to assess their size and optical properties respectively. Cell toxicity was investigated using MTT and glutathione assay. Intracellular uptake and cell distribution was imaged using confocal laser scanning microscopy. Quantum dots CdSe/ZnS_TOPO and quantum dots-capped with bisphosphonates (CdSe/ZnS_BIPs) were successfully synthesised. TEM showed the size and morphology of the quantum dots to be 5.0nm ± 0.5nm with spherical shape. The stabilised quantum dots-capped bisphosphonates have showed good dispersion in aqueous solutions and retained their fluorescent characteristics. The cytotoxicity studies showed a concentration dependant effect on cell viability induced by quantum dots CdSe/ZnS_BIPs. This project demonstrates reproducible and less hazardous method in the synthesis of quantum dots CdSe/ZnS and a novel approach in producing water dispersible quantum dots through ligand exchange with bisphosphonate groups. However, further work will involve determine the stability in aqueous solutions and to establish a purification protocol for cell and in vivo bioimaging of CdSe/ZnS_BIPs. [1] B.O. Dabbousi, J. Rodriguez-Viejo, F.V. Mikulec, J.R. Heine, H. Mattousi, R. Ober, K. F. Jensen and M.G. Bawendi. (CdSe)ZnS Core-Shell Quantum Dots: Synthesis and Characterization of a Size Series of Highly Luminescent nanocrystallites. J. Physicals Chemistry B., 1997, 101 (45), pages 9463-9475 [2] Kimihiro Susumu, Bing C Mei & Hedi Mattousi. Multifunctional ligands based on dihydrolipoic acid and polyethylene glycol to promote biocompatibility of quantum dots. Nature Protocols, Vol. 4 No. 3 (2009) page 424-436

NANO-110 Development trachea with bronchi for paediatric application from novel bioabsorbable nanocomposite scaffold and stem cells technology G.Z. Teoh1, C.Crowley1,2, M. Birchall1,2, A.M.Seifalian1

University College London (1UCL Centre for Nanotechnology and Regenerative Medicine, Division of Surgery and Interventional Sciences, 2UCL Ear Institute, Royal National Throat, Nose & Ear Hospital), London, U.K.

Tracheal defects like congenital tracheal stenosis (CTS) and prolonged intubation following premature birth have resulted in an unmet clinical need. Advances in stem cell technology, tissue engineering and material sciences have inspired the development of our resorbable, nanocomposite trachea and bronchi scaffold. A trachea and bronchi scaffold was designed and constructed using our novel, resorbable nanocomposite polymer, Polyhedral Oligomeric Silsesquioxane Poly(ε-caprolactone) Urea Urethane (POSS-PCL), integrated with Polyhedral Oligomeric Silsesquioxane Poly(carbonate-urea) urethane for additional mechanical strength. Material characterisation studies included polymer viscosity, tensile strength, suture retention and contact angle. Bone marrow derived mesenchymal stem cells (bmMSCs) and human tracheobronchial epithelial cells (HBECs) were isolated, passaged and cultured on POSS-PCL for 14 and 8 days respectively and the metabolic activity was assessed by Alamar Blue® assay. Quantum dots conjugated to RGD peptides and anti-collagen type I antibody were then used to examine cell migration throughout the scaffold. POSS-PCL exhibited good mechanical properties and the relationship between the casted and coagulated polymer layers was comparable to the relationship between the cartilaginous U-shaped rings and interconnective cartilage of the native human trachea. Good suture retention and thickness of the scaffold was also achieved. Isolation and in vitro culture protocols for bmMSCs and HBECs were successfully optimised. Cell attachment and a significant, steady increase in proliferation was observed, as illustrated by two-way ANOVA (bmMSC: p=0.001; HBEC: p=0.003). Antibody-quantum dot imaging illustrated cell penetration throughout the scaffold. SEM images confirmed the attachment and proliferation of both cell lines as well. Our mechanically viable synthetic scaffold provides a conducive environment for bmMSC and HBEC attachment and proliferation demonstrating its potential as a tissue-engineered solution to the increasing number of tracheal anomalies and pathologies in clinics. NANO-111 Functionalized Carbon Nanotubes Enhance Ex Vivo Expansion of Hematopoietic Stem and Progenitor Cell in Human Umbilical Cord Blood S Bari1*, PPY Chu1,2*, A Lim1, X Fan3, FPH Gay1, RM Bunte4, L Zheng5, S Li6, GNC Chiu7**, WYK Hwang1,2,6**

1Department of Hematology, Singapore General Hospital, Singapore 2Singapore Cord Blood Bank, Singapore 3Department of Clinical Research, Singapore General Hospital, Singapore 4Office of Research, Duke-NUS Graduate Medical School, Singapore 5Singapore Experimental Medicine Center, SingHealth, Singapore 6Cancer & Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore 7Department of Pharmacy, National University of Singapore, Singapore

The clinical application of umbilical cord blood (UCB) for hematopoietic stem cell transplantation (HSCT) is often hampered by low cell dose and reduced viability from frozen cell sources. These problems could be partly resolved by using cytokine-mediated stromal co-culture ex vivo expansion system, as previously shown by us and others. In this current study, we have explored the use of single-walled carbon nanotubes functionalized with carboxylic acid (f-SWCNT-COOH) as a scaffold, and demonstrated its ability to support viability & ex vivo

expansion of freeze-thawed, non-enriched hematopoietic stem and progenitor cells (HSPC) in UCB–mononucleated cells (UCB-MNCs). UCB-MNCs cultured in serum free media with a basal cocktail were exposed to commercially available f-SWCNT-COOH at various concentrations. UCB-MNC cultures without the f-SWCNT-COOH served as the control. The effects of f-SWCNT-COOH treatment were characterized based on UCB-MNC viability, phenotypic-based stem and progenitor cell number and functional in vitro and in vivo assays. In a concentration-dependent manner, 1.0 mg/mL f-SWCNT-COOH was found to support the viability of the CD45+ as assayed using the Annexin-V/7-AAD stains at 72 hours (p<0.0001; n=48) post-culture with a decrease in the proportion of CD45– population (p<0.0001; n=48) and the observed phenomenon being independent of cytokine stimulation. In an 11-day, cytokine supplemented expansion system, f-SWCNT-COOH ex vivo enhancement effect was observed in viable nucleated CD45+ cells (1.5-fold; p<0.05; n=18); hematopoietic progenitor cells (HPCs: CD45+CD34+CD38–) (54-fold; p<0.0001; n=24); hematopoietic stem cells (HSCs: CD45+CD34+C38–CD90+) (80-fold; p<0.0001; n=15) and common myeloid progenitors (CMPs: CD45+CD34+CD13+CD33+) (25-fold; p<0.001; n=9) compared to control. Furthermore, UCB-MNC cultured with f-SWCNT-COOH showed 5- and 211-fold enhancement effect in functional colong forming unit-GM (n=81) and GEMM (n=45) assay, respectively (p<0.0001). Trasplantation of f-SWCNT-COOH expanded UCBs, particularly at 1x108cells/kg, in sub-lethally irradiated non-obese diabetic (NOD) – severe combined immunodeficiency (SCID) gamma mice (NSG) resulted in better human chimerism in the murine peripheral blood (PB) as early as 8 weeks post-transplantation compared to control (n=11; p=0.16). Bone marrow (BM) analysis of the NSG mice transplanted with 2x108cells/kg also showed that f-SWCNT-COOH expanded graft exhibited better human multilineage reconstitution (human hematopoetic cells, progenitors and B-lineage) after 16-weeks of transplantation (n=5; p=0.22). It must be noted that concurrent transplantation of the same non-expanded UCB-MNCs graft resulted insignificant difference of human chimerism in the NSG mice PB and BM (n=8) compared to the expanded grafts (±f-SWCNT-COOH). However, the non-expanded graft resulted in higher incidence of graft-versus-host disease (GVHD) in the mice even with immunosuppresion that translated to higher mortality rate (54.3%; n=35) compared to the f-SWCNT-COOH expanded graft (14.6%; n=55) and control (13.2%; n=38) (p<0.01) over 120 days. This observation could be attributed to the significantly higher proportion of developing immune cells (CD45+CD7+) in the non-expanded grafts compared to the expanded grafts (±f-SWCNT-COOH) (p<0.001). Intravenously administered f-SWCNT-COOH into the NSG mice (n=22) revealed no evidence of acute or chronic toxicity over 6 months as compared to saline control (n=5), assessed by parameters such as body weight, survival, and histological analysis of major organs. We have demonstrated a novel application of carbon nanotubes in the ex vivo expansion of UCB-MNCs. Compared to other reported bio-scaffolds, f-SWCNT-COOH was able to successfully enhance the ex vivo expansion of HSPCs in UCB-MNCs without the prior need of cell selection and yet maintain the in vivo functional ability of the SCID-repopulating cells. NANO-112 Investigating the Delivery of m-THPC incorporated in Pegylated PLGA Nanoparticlesfor in vivo PDT. M.J. Bovis*1, J.H. Woodhams1, M. Rojnik2, P. Kocbek2, J. Kos2, S.G. Bown1, A.J. MacRobert1,M. Loizidou3.

1National Medical Laser Centre, Division of Surgery & Interventional Science and Institute of Biomedical Engineering, University College London, W1W 7EJ, UK. 2Faculty of Pharmacy, University of Ljubljana, SI-1000, Slovenia. 3Royal Free Hospital, Dept. of Surgery, University College London, NW3 2Q, UK.

Photodynamic therapy (PDT) is a minimally invasive procedure effective in the treatment of cancer and certain non-malignant diseases. In this in vivo study, we investigated the biodistribution of a potent photosensitiser, meta-tetra(hydroxyphenyl)chlorin (m-THPC), in its standard formulation (Foscan®) and when encapsulated in both pegylated poly(lactic-co-glycolic acid) (PEG-PLGA) and non-pegylated PLGA nanoparticles (NPs) at selected time intervals. These data were compared to their relative photodynamic efficacy in a rat tumour model. Foscan

and PLGA m-THPC NPs were injected i.v. into Hooded Lister (HL) rats (0.3 mg kg-1 m-THPC). Tissue samples (tumour, liver, spleen, kidney, lung, skin, muscle and blood plasma) were removed to examine m-THPC accumulation through chemical extraction and fluorescence microscopy techniques at 24 and 72 h. A methylcholanthrene-induced fibrosarcoma cell line (MC28), syngeneic to HL rats, was chosen as a suitable tumour model. In a separate group of animals, tumours (10 mm in diameter) were irradiated with red laser light interstitially from a 652 nm diode laser. A 400 µm bare-cleaved tip optical fibre was inserted into the tumour capsule, mimicking interstitial clinical PDT. Drug-light intervals (DLIs) were identical to those times chosen for pharmacokinetic analysis. Tumours were resected post-mortem and processed by routine histological methods. Necrosis was calculated as a percentage of the total tumour surface area from a series of 4 micron sections to assess PDT efficacy. m-THPC in PEG-PLGA NPs appeared to accumulate less in normal tissues at 24 and 72 h than when encapsulated in PLGA NPs. This can be attributed to the protective PEG coating, which helps prevent recognition and uptake of the NP into macrophages. Using Foscan, m-THPC was observed at much greater concentrations in the kidneys, which is likely due to NP excretion via the hepatobiliary pathway. Both sets of NPs demonstrate a greater uptake of m-THPC in tumour tissue with PLGA NPs in comparison to standard Foscan over 72 h (~2-3 fold). However despite this greater accumulation in tumour tissue, there appeared to be limited differences in PDT efficacy between formulations and DLIs. Using fluorescence lifetime spectroscopy of freshly excised liver, comparable lifetimes were observed for the NPs and Foscan delivery at 24 h, which demonstrates that the NPs had by then degraded and released the m-THPC. PDT studies using m-THPC encapsulated in PLGA NPs show encouraging results. Use of these PLGA NPs additionally permits the attachment of surface ligands, capable of recognising tumour-specific receptors to further improve m-THPC uptake and PDT efficacy. NANO-113 Bifunctional Ag3PO4/TiO2/Graphene Composite Materials with Synergistic Antibacterial Activities and Enhanced Photocatalytic Performance Dr. X.F. Yang1, Dr. H.Y. Cui2, J.L. Qin1

1School of Materials Science and Engineering, Jiangsu University, Zhenjiang, 212013, P. R. China. 2School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, P. R. China.

Biological and environmental pollutions have been found to be present widely in the environment and have long been recognized as a threat to human health as well as to the earth's ecosystems. A variety of bacteria and toxic pollutants in aqueous and gaseous media are considered to be largely responsible for human diseases including cancer, inherited disorders and mental disability. The elimination of bacteria and toxic pollutants, especially persistent and volatile organic compounds, in the environment simultaneously has been a great challenge. Photocatalysis has been proven to be an effective way for the degradation of hazardous organic pollutants. Most recently, silver orthophosphate (Ag3PO4) was report to possess significant visible-light-driven photocatalytic degradation performance towards organic dyes under visible-light irradiation. However, it is undeniable that the cost of silver-containing raw material is too high to afford for large-scale production, as-prepared Ag3PO4 samples normally possess irregular polyhedral microstructures and present poor solubility, its photocatalytic activity has also been proved to be unstable. Considering its excellent photocatalytic performance and potential antibacterial activities, we develop a low-cost hydrothermal method for the preparation of bifunctional Ag3PO4/TiO2/graphene composite materials where the size as well as morphology of Ag3PO4 particles are tailored by the added graphene oxide and the high cost of reactants can be reduced due to the addition of inexpensive TiO2. The generation of heterogeneous Ag3PO4/TiO2 structures and the in-situ transformation of graphene oxide to graphene could be simultaneously achieved in the hydrothermal process. The composition, particle size, structure and morphology of the obtained composite materials are fully characterized. The photocatalytic results show that composite materials exhibit highly active degradation rate of organic pollutants. Moreover, three methods including inhibition zone, minimum inhibitory concentration (MIC)/minimum bactericidal concentration

(MBC), time-dependent bactericidal activity calculated by residual bacteria count were employed to investigate antibacterial performance of composites against seven types of bacteria. Our findings show broad-spectrum antibacterial activity of composites that possibly caused by synergistic antibacterial effects between GO sheets, TiO2 and Ag3PO4. Our findings offer a great opportunity for the the low-cost, large-scale production of bifunctional materials with highly active photocatalytic properties and excellent antibacterial activities for potential applications in different fields. [1] Z.G. Yi, J.H. Ye, N. Kikugawa, T. Kako, S.X. Ouyang, H. Stuart-Williams, et al., Nat. Mater., 9, 559 (2010). [2] Y.P. Bi, S.X. Ouyang, N. Umezawa, J. Cao, J.H. Ye, J. Am. Chem. Soc., 133, 6490 (2011). [3] W.F. Yao, B. Zhang, C.P. Huang, C. Ma, X.L. Song, Q.J. Xu, J. Mater. Chem., 22, 4050 (2012).

NANO-114 Novel materials for liver directed therapy J. Schnekenburger1; N. Schmitt2, K. Hoffmann3, J. Bruder4, C. Rommel1, F. Schröter5, S. Heine3, A. Zibert2, H. Schöler4, B.J. Ravoo5, ,M. Schönhoff3, H.H.J. Schmidt2

1Biomedical Technology Centrum;2Clinic for Transplantation Medicine; 3Institute of Physical Chemistry; 4MPI for Molecular Biomedicine; 5Organic Chemistry Institute; University of Münster, Germany

Functionalized materials have an enormous potential for medical applications, e.g. for the development of cell based therapy. Although hepatocytes are currently used in the clinic for therapy of liver disease, the availability of the cells and the specific delivery to the target organ is limited, due to organ shortage, the reduced propagation of hepatocytes in standard 2D tissue culture, the lack of efficient differentiation of hepatocyte-like cells from stem cell precursors, like iPS (induced pluripotent stem) cells, and by degradation of cells after transplantation. Thus, molecular, physical, and preclinical characterization of the interaction between functionalized materials and hepatocytes/stem cells are highly warranted. (1) A variety of organic and biologic polymers (n=14) were combined by electrostatic forces to built highly controllable 2D and 3D layers that are suited as a surface for cultivation of the fragile hepatocytes. Combinations of PLO (Poly-L-ornithine hyrobromide)-PGal (Poly-L-galacturonic acid) and PArg (Poly-L-arginine hydrochlorid )-PGal, amongst others, where found to sustain proliferation of a hepatocyte cell line whereas higher combinations of polymers, e.g. (PLO-PGal)3-PLO showed inhibitory effects. (2) Layer-by-layer assembly methodology was transferred to encapsulation of hepatocytes. Using flow cytometry analysis cells were observed to increase light scattering properties that were highly specific for individual polymer. PLO-PGal-PLO encapsulation significantly increased FSC (675±45) and SSC (495±70) mean fluorescence values as compared to untreated cells (665±45 and 142±9, respectively). (3) Targeting of hepatocytes to defined nanostructured surfaces was investigated by micro patterning technology using defined peptide sequences that were shown to bind hepatocytes. Binding of hepatocytes was assessed by Electric Cell-substrate Impedance Sensing (ECIS). Ultimately, novel materials that modulate primary cells including stem cells will be evaluated in animal models of liver disease.

NANO-115 Diamond Nanocones for Improved Osteoblastic Differentiation E.Y.W. Chong1, C.Y.P. Ng2, V.W.Y. Choi2, Y. Yang2, W.J. Zhang2, K.M.C. Cheung1, K.W.K. Yeung1, X.F. Chen2, K.N. Yu2

1 Department of Orthopaedics and Traumatology, University of Hong Kong, Hong Kong 2 Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong, Hong Kong

The diamond nanocone growth was achieved through depositing a thin layer of diamond film on a silicon substrate followed by bias-assisted reactive ion etching [1]. The nanocones are highly densely packed and the

height varies from 200 nm to 1 m. The tip diameter of the nanocones is below 10 nm. We hypothesize that these nanocones can mechanically disrupt cell membranes which will aid the delivery of molecules into cells but without causing irreversible damage to cells. To confirm our hypothesis, we tested the nanocone aided delivery of cell differentiation agent into osteoblast cells. In the experiments, a total of 25,000 MC-3T3 cells in 1 ml differentiation medium which contained 50 µg/ml ascorbic acid (Sigma, USA) and 20 mM β-glycerol phosphate (Sigma, USA) were pipetted and applied to a nanocone patch at 3 m/s. The cell suspension was collected and the cell treatment procedure was repeated 15 times. The treated cells were then transferred into 24 well plates. Equal concentration of cells without being treated with nanocones was used as control. The cells were cultured at 37oC with 5% carbon dioxide and 95% humidified air. The differentiation medium was changed every 2-3 d. At designated time points, i.e., 3rd, 7th and 14th days, alkaline phosphatase (ALP) activity, one of the typical markers for osteoblastic differentiation, was studied with alkaline phosphatase activity assay kit (Stanbio Alkaline Phosphatase LiquiColor®). Enzymatic activity was normalized to total protein concentration using bovine serum albumin. ALP activity was expressed as U/mg protein. The cultured MC-3T3 cells treated with nanocones showed significantly higher ALP activity on the 3rd and 7th days than the control while the activity was lower on the 14th day. Osteoblastic differentiation was enhanced at the early time points, indicating that early bone formation was potentially promoted [2]. This technique provides a very simple approach to achieve delivery of molecules to large number of cells. [1] C. Wild, R. Kohl, N. Herres, W. Muller-Sebert, and P. Koild, Diamond Relat. Mater., 3, 373 (1994). [2] M. Sila-Asna, A. Bunyaratvej, S. Maeda, H. Kitaguchi, N. Bunyaratavej, Kobe J Med Sci., 53, 25 (2007).

NANO-116 Cadmium Free Quantum Dots as a Novel Fluorescence Probe for In Vivo Sentinel Lymph Node Imaging

1Elnaz Yaghini, 2Helen Turner, 2Imad Naasani, 2Lesley Smith, 1Alexander J MacRobert

1National Medical Laser Centre, Division of Surgery and Interventional Science and the Institute of Biomedical Engineering, University College London, London, UK 2Nanoco Technologies Ltd. 46 Grafton Street, Manchester, UK

Fluorescence imaging is an alternative to the current method (blue dye and radiocolloid) of sentinel lymph node (SLN) mapping in breast cancer. Quantum dot nanoparticles (QDs) with unique optical properties could be used for this purpose. However, the widely used cadmium (Cd) containing QDs may induce toxicity, which could be avoided by replacing Cd with more benign elements, such as indium (In). Our aim was to develop new biocompatible cadmium-free (CF) QDs and study their potential for SLN mapping under in vivo condition. Water soluble indium-based QDs were administered subcutaneously in the anterior paw of healthy Hooded Lister rats. Animals were sacrificed at various time intervals and different organs including axillary lymph node (ALN) and thoracic lymph node (TLN) were harvested for the pharmacokinetic studies. The QDs biodistribution was

investigated by elemental analysis of Indium (In) using the inductively coupled plasma mass spectroscopy (ICP-MS). Ex vivo fluorescence imaging of the harvested tissues were recorded with a CDD camera. Frozen section fluorescence microscopy of tissues was also carried out. Using ICP-MS QDs were detected in ALN and TLN as early as 10 min after injection. The overall level of QDs in other tissues remained low. Additional evidence for the uptake of QDs in ALN and TLN was obtained by ex vivo fluorescence imaging and frozen section fluorescence microscopy of the harvested tissues. In summary, we report the development of biocompatible CF QDs with an excellent quantum yield (QY) for in vivo imaging. Effective and rapid detection of sentinel lymph node using fluorescence imaging of QDs could make them attractive for biological and diagnostic applications. NANO-117 Singlet Oxygen Generation from Water Soluble Quantum Dot-Photosensitiser Nanocomposites and their Potential for Photodynamic Therapy Elnaz Yaghini1, Alexander M Seifalian2, Alexander J MacRobert1

1National Medical Laser Centre, Division of Surgery and Interventional Science and the Institute of Biomedical Engineering, University College London, London, UK 2 Centre for Nanotechnology and Regenerative Medicine, Division of Surgery and Interventional Science, University College London, London, UK.

Photodynamic therapy (PDT) is a minimally invasive therapeutic modality approved for the clinical treatment of several types of disease. We studied the use of quantum dot (QD) for PDT by developing quantum dot-photosensitiser (QD-PS) nanocomposites. Generation of singlet oxygen (1O2) could be achieved via indirect excitation of photosensitiser (PSs) through Förster resonance energy transfer (FRET) from QDs to PSs. Water soluble QD-PSs nanocomposites were developed where QDs were attached to phthalocyanine (AlPcS2a) photosensitisers through electrostatic interaction. Tat-peptide conjugated QD620 and QD680 and, their unconjugated counterparts were selected as energy donors. The amphiphilic disulphonated aluminium phthalocyanine (AlPcS2a) photosensitiser was utilised as the energy acceptor. Following the excitation at 488 nm which is near the minimum of PSs absorption spectrum, a substantial decrease in QDs photoluminescence (PL) was observed along with enhancement in PSs fluorescence due to FRET between these complexes. Further evidence for the occurrence of FRET was obtained by fluorescence lifetime measurements, where a substantial shortening of the QD donor lifetime was measured in the presence of the PSs in a concentration and time dependent manner. The efficiency of these nanocomposites to generate 1O2 was shown by direct measurement of 1O2 phosphorescence at 1270 nm. The 1O2 quantum yield (QY) was determined by comparing intensity of 1270 nm signal of 1O2 with that of Rose Bengal, as a reference. In summary, this study revealed that the QD can be used to sensitise the PDT agent through FRET process which in turn resulted in the generation of 1O2 that could be used for PDT cancer therapy. NANO-118 Oligonucleotide-decorated carboxyl-modified PVP nanogels: a new platform for diagnosis and gene-therapy C. Dispenza1, G. Adamo2, M.A. Sabatino1, M.L. Bondì3, M.P. Casaletto3, S. Rigogliuso2, G. Ghersi2

1Dipartimento di Ingegneria Chimica, Gestionale, Informatica, Meccanica, Università degli Studi di Palermo, Edificio 6, Italy. 2Dipartimento di Scienze e Tecnologie Molecolari e Biomolecolari, Università degli studi di Palermo, Viale delle Scienze, Italy. 3Consiglio Nazionale delle Ricerche, Istituto per lo Studio dei Materiali Nanostrutturati, U.O.S. Palermo, via Ugo La Malfa, Palermo.

Nanogels, or nanoscalar chemically crosslinked polymer networks, were generated by simultaneous crosslinking of Poly(N-vinyl pyrrolidone) (PVP) and grafting of acrylic acid (AA) monomer through e-beam irradiation of their

aqueous solutions. The yield of the process is very high and no separation or complex purification procedures are required since no recourse to surfactants or organic solvents is made. The administered irradiation dose (40 kGy) imparts also sterility to the obtained nanogels in the form of aqueous dispersions. Control of particle size and size-distribution can be achieved by proper selection of the irradiation conditions. In particular, we obtained nanogel variants with average hydrodynamic diameters in the range 30 – 100 nm with PDI lower than 0.3 and negative surface charge density. Confirmation of the molecular architecture and detection of the surface composition have been sought through complementary spectroscopic techniques, such as FTIR and XPS. Preliminarily to any biological evaluation or bio-conjugation, absence of cytotoxicity was tested through MTT assay. Studies on nanogel viability thought cell compartments, carried out with fluorescent PVP nanogels variants incubated for different times in cell cultures and observed by confocal microscopy, showed cytoplasmic distribution of nanogels and preferential localization of around the nuclei during the first 7 hours and progressive disappearance during the following 17 hours. Nanogels secretion as waste products could be hypothesized. The availability of accessible carboxyl groups for subsequent facile bio-conjugation reactions makes this family of functional nanogels very promising for decoration with oligonucleotides. As a proof of concept, a fluorescent oligonucleotide semi-helix was conjugated through an amino group attached to the terminal 3’. The recognition ability of the conjugated forward semi-helix toward the reverse semi-helix was proved by spectrofluorimetric readings using a reverse oligo carrying a fluorescence quencher. NANO-119 Effect of lipid coating on the interaction between silica nanoparticles and membranes D.B. Tada1,a, E. Suraniti2, L.M. Rossi1, C.S. Oliveira1,b,, T.C. Tumolo1; R. Calemczuk2, T. Livache2, M.S. Baptista1 1 Instituto de Química, Universidade de São Paulo, SP, Brazil 2CREAB – SPRAM, UMR 5819, INAC/CEA-Grenoble, France aPermanent address: ICT, Universidade Federal de São Paulo, São José dos Campos, SP, Brasil. bPermanent address: Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS, Brasil.

Lipid coating is a method highly used to improve the biocompatibility of nanoparticles (NPs), even though its effect on the NP properties is still object of investigation. Herein, silica NPs containing methylene blue[1], which is a photosensitizer used in a variety of biomedical applications, were coated with a phospholipid bilayer. These NPs (bare or lipid-coated) generate singlet oxygen and induce cell death with similar efficiencies. However, the cytosolic distribution of these NPs was totally different. Lipid-coated NPs were distributed through the cell cytoplasm whereas bare NPs were detected only in some vacuolar regions within the cells. Since cellular uptake and cytolocalization are influenced by NP adsorption on cell membranes, the interaction of lipid-coated and bare NPs were studied on a membrane mimic, i.e., Hybrid Bilayer Membranes (HBMs) made of different compositions of negatively charged and neutral lipids. Interactions of bare and lipid-coated NPs with HBMs were analyzed by Surface Plasmon Resonance Imaging[2]. Bare NPs presented high adsorption and aggregation on HBMs independently of the surface charge. Conversely, lipid-coated NPs presented less aggregation on the membrane surface and the adsorption was dependent on the charges of the NPs and of the HBMs. Our results indicate that NPs aggregation on the membrane surface can be modulated by lipid coating, which affects the cytosolic distribution of the NPs. [1] D.B. Tada, L.L.R. Vono, E.L. Duarte, R. Itri, P.K. Kiyohara, M.S. Baptista, L.M. Rossi. Langmuir, 23, 8194 (2007). [2] E. Suraniti, T. Tumolo, M.S. Baptista, T. Livache, R. Calemczuk. Langmuir, 23, 6835 (2007).

NANO-120 Preparation of Nanostructured Polymer Surface for Control of Cell Behavior T. An*1, H. Nam2, and G. Lim2,3 1Department of Mechanical Design Engineering, Andong National University, Andong, Gyungbuk, Korea. 2Department of Integrative Bioscience and Biotechnology, POSTECH, Pohang, Gyungbuk, Korea. 3Department of Mechanical Engineering, POSTECH, Pohang, Gyungbuk, Korea.

Most cells derived from animal tissues have anchorage-dependency. And cell and tissue behavior can be profoundly affected by microenvironment. In particular, living cells are exquisitely sensitive to the local micro- and nanoscale structure of the growth substrate [1]. Therefore, studies of the interaction between cell and topological structure are great importance. Recently, various fabrication methods have been applied to fabricate micro/nano structure on the cell culture substrate [2,3]. In this research, we demonstrated simple fabrication methods of porous PMMA micro/nano-surface for cell culture substrate. Porous PMMA substrate had nanopapilla structure with a size of 100 ~ 300 nm. To fabricate porous PMMA substrate, PMMA film surface was treated by organic solvents to dissolve and was pressed by micro mold for patterned microstructure. Finally, PMMA film treated insoluble solvents and nanopailla structure was formed because organic solvent was quickly removed by insoluble solvent form PMMA film. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2012012805). [1] M. Stevens, J. George, Science, 310, 1135 (2005) [2] D.H. Kim, H. Lee, Y. K. Lee, J. M. Nam, A. Levchenko, Adv. Mater, 22, 4551 (2010) [3] N. M. Alves, I. Pashkuleva, R. L. Reis, J. F. Mano, Small, 6, 2208 (2010)

NANO-121 Fabrication of anti-bacterial nanofiber containing Ag-nanoparticles T. An*1 1Department of Mechanical Design Engineering, Andong National University, Andong, Gyungbuk, Korea.

Silver nanoparticles have superior anti-biotic activity[1]. So dressings with superior antibiotic activity will be manufactured by producing nanofiber dressings containing silver nanoparticles [2]. Nanofiber dressings containing silver nanoparticles can not only minimize the adverse effect of antibiotics by reducing the use of antibiotics but also have superior effectiveness when compared to existing dressings using silver-ion drugs. By manufacturing nanofibers by controlling the size and concentration of silver nano-particles, the antibiotic activities by different sizes and concentrations of silver nano-particles will be examined to manufacture nano-fiber dressings with optimized effectiveness to the type of the wound. In this research, we demonstrated biodegradable anti-biotic nanofiber which have a diameter of 100 ~ 300 nm and contain silver nanoparticles. Silver nanoparticles of the size 20 ~ 50 nm have been synthesized by liquid phase reduction method. Biodegradable nanofiber was fabricated by electrospining method. To control the concentration of silver nanoparticles on the nanofiber surface, PLGA solutions with different concentration of silver nanoparticles were used for electrospinning process. Because the environment for the wound recovery changes according to the type of the wound, anti-biotic nanofibers with various concentrations of silver nanoparticles can be used materials of optimized wound dressing for each wound. This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education, Science and Technology(2012012805) [1] G. A. Sotiriou and S. E. Pratsini, Environ. Sci. Technol., 44, 5649 (2010) [2] X. Xu and M. Zhou,Fiber and polymer, 9, 685 (2008)

NANO-122 Spatial 3D Modelling of Colorectal Cancer: A Tissue Engineering Approach T. Magdeldin1, A Nyga1,M. Loizidou1,U. Cheema1,2 University College London (1UCL Centre for Nanotechnology and Regenerative Medicine, Division of Surgery and Interventional Sciences, 2UCLInstitute of Orthopaedics and Musculoskeletal Science) London, UK.

Most pre-clinical cancer research is carried out in reductionist 2D cell culture or animal models. Although 2D in vitro cultures are controllable, amenable to molecular manipulation and invaluable for mechanistic investigations; they do not provide an accurate representation of the complexity of in vivo interactions. The use of animal models, while also important, are subject to interspecies differences that do not always mimic the human scenario. 3D cell cultures bridge the gap between conventional 2D cell culture studies and model organisms. We have developeda spatially accurate 3D in vitro model of colorectal cancer. Using tissue engineering techniques such as plastic compression, colorectal cancer cells (HT29) and collagen type I were used to construct a dense, artificial cancer mass (ACM). To mimic the tumour microenvironment, the ACM was positioned within stromal cell populated uncompressed collagen containing endothelial cells (HUVECs) and fibroblasts (3T3). Characterization of the model included investigations of: cell growth (Alamar Blue®), the presence of hypoxia, a key feature of solid cancers (O2 probes), cell morphology and immunofluorescence (VEGF). Cell survival within the matrix was characterized up to 21 days. Cancer cells formed cell aggregates similar to tumour spheroids in vivo and invaded outwards from the ACM. O2 levels within the core of the ACM decreased to ~10mmHg within 4 hours, compared to 70mmHg in the surrounding matrix (p<0.05).Immunofluorescence confirmed VEGF production by cancer cells at the invading edges of the ACM. Our 3D model exhibits fundamental characterstics of the tumour microenvironment (spatial organization of cell types, tumour invasion, hypoxia and VEGF expression) and thus demonstrates realistic potential as an alternative to animal testing. The model is currently under development for therapeutic and imaging purposes. NANO-123 The Kinetics of Anesthetic Enhanced Membrane Fusion in DOPC Liposomes T.T. Nguyen1, D.T. Cramb1

1University of Calgary, Calgary, Alberta, T2N1N4, Canada.

Lipid vesicles (liposomes) are attractive drug and gene delivery vehicles in the field of nanomedicine, due to their small size, biocompatibility, non-toxic behaviour and ability to transport both hydrophobic and hydrophilic drugs.[1] However, the mechanism for drug release/uptake once the liposome contacts the cell surface remains speculative. Liposomes can be taken up by endocytosis, phagocytosis, or fusion could occur between the liposome and the plasma membrane, releasing the vesicle contents (drug) into the cell.[2]. The mechanism of membrane fusion has been under debate for many years, it is now generally accepted that both proteins and lipids play an important role in the process of fusion. There have been studies conducted using pure lipids membranes that have also illustrated fusion[3,4], which indicates that proteins are not necessarily required for fusion to occur. It has been previously shown that a general anesthetic (halothane) can be used as a fusion agent in unilamellar vesicles of dioleoylphosphatidylcholine (DOPC).[3] This study investigates the kinetics of vesicle association and lipid mixing in DOPC liposomes induced by halothane. Fluorescence based assays were utilized to report on different stages within the membrane fusion mechanism. By understanding the mechanism of fusion, better fusion enhancing agents can be developed for use in liposomal drug delivery. [1] T.C. Yih, and M. Al-Fandi, J. Cell. Biochem., 97, 1184 (2006).

[2] A.S. Ulrich, Bioscience Rep., 22, 129 (2002). [3] J.L. Swift, A. Carnini, T.E.S. Dahms, and D.T. Cramb, J. Phys. Chem. B, 108, 11133 (2004). [4] M.E. Haque, T.J. McIntosh, and B.R. Lentz, Biochemistry, 40, 4340 (2001).

NANO-124 Synthesis and characterization of Ni nanoparticles S.H. Masunaga1, R.F. Jardim1

1Instituto de Física, Universidade de São Paulo, São Paulo, SP, Brazil

Magnetic nanoparticles (NPs) have been intensively studied due to their potential application in biomedicine. For instance, magnetic NPs can be used as MRI contrast agents, magnetic carriers for drug delivery and tagging biological entities. Ni NPs embedded in SiO2/C matrix were prepared through a method in which silicon oxianions and the metal cation (Ni) are immobilized within a polymeric matrix based on polyester with Ni concentrations up to 20 % wt. Initially, citric acid was dissolved in ethanol and Si(OC2H5)4, then Ni(NO3)2·6H2O was added and mixed for homogenization. Finally, ethylene glycol was added to promote the polymerizing reaction. The resulted solid resin was heat treated at 300 °C for 3 h, ground in ball mill, and then pyrolyzed in N2 atmosphere at 500 °C. The pyrolysis of the precursor resulted in a CO/CO2 rich atmosphere, which together with the N2 flux, promote the reduction of the Ni citrate, resulting in metallic Ni NPs. This is an easy and quick method which allows a production of a large amount of sample at low cost and provides a good control of particle size distribution. The images from TEM indicate that the Ni NPs are homogeneously distributed within the matrix, with no sign of agglomeration, and have nearly spherical shape, with median particle size of 5 nm and narrow size distribution. We have performed a systematic study of magnetic properties by means of ac and dc magnetic susceptibility and hysteresis loops measurements. The temperature dependence of magnetization, obtained after zero-field-cooling process, exhibits a maximum related to the blocking temperature (TB) of the system and a superparamagnetic behavior above TB. The same magnetic properties were observed after several years of the samples exposed to air (at least for 3 years), indicating that Ni NPs are well protected inside SiO2/C matrix. Furthermore, from the magnetic properties of the samples, the dipolar interactions between Ni nanoparticles can be considered negligibly small or very weak. All these properties make these samples suitable for biomedical applications as nano-vehicle delivery systems because Ni NPs can be manipulated by an external magnetic field. NANO-125 Enhancement of Cancer Therapy Efficacy by Herceptin-conjugated and pH-Sensitive Nanocapsules with Simultaneous Encapsulation of Hydrophilic and Hydrophobic Compounds C.S. Chiang, S.S. Hu, S.Y. Chen Department of Materials Science and Engineering, National Chiao Tung University, Hsinchu 300,Taiwan

Recent advance in nanotechnology has produced a number of drug delivery systems to improve therapeutic efficacy and reduce side effect. Smart combination of drugs in nanocarriers can better modulate cell signaling network to maximize therapeutic effect and reduce drug resistance. Despite these recent advances in nanocarrier engineering, technological challenges in encapsulating multiple therapeutic compounds in one NP still exist. A double emulsion nanocapsule (DENC) stabilized by Single-Component PVA Polymer with magnetic nanoparticles (MNPs) can be fabricated through two-steps double emulsion processes and is capable of carrying both hydrophilic and hydrophobic anti-cancer drugs (PTX/Dox) with desirable dosage in one compact nanocapsule[1]. The composite shell of DENC was composed of pH-sensitive polymer (PMASH) and MNPs, which

exhibited two major functions: (1) protecting cargos from leakage and (2) controlling the released amounts of PTX/Dox in the intracellular acidic pH environment. Furthermore, the DENC was conjugated with targeting antibody, Herceptin® to actively delivery the nanocapsules to treat the Her2/Neu overexpressing breast cancer. The enhanced cell and tumor uptake of Her2/Neu overexpressing breast cancer can be clearly observed in vitro and in vivo. In addition, it has been shown that co-delivery of paclitaxel and doxorubicin using the DSNC polymeric nanoparticles can suppress breast tumor growth in a mouse model more efficiently than the delivery of either compound alone. Together with pH-sensitive approach and bio-targeting, the DENC developed in this study may offer a potential treatment approach of dual targeting and smart intracellular controlled release for advanced cancer therapy. [1] S.S. Hu, B.J. Liao, P.J. Chen, I.W. Chen and S.Y. Chen, J. Adv.Mater, 24,27, 3727 (2012).

NANO-126 Multifunctional Large Pore Mesoporous Silica Nanoparticles as a novel gene carrier S.B. Hartono1, W.Gu1, M. Yu 1, J. Yang 1, E. Strounina 2, G. Q. Lu1, S. Qiao1, C. Yu1

1 ARC Centre of Excellence for Functional Nanomaterials, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, QLD 4072, Australia 2 Centre for Advanced Imaging, The University of Queensland, St Lucia, QLD 4072, Australia

Small interfering RNA (siRNA) based therapeutics for cancer treatments have gained considerable attention due to the high potency of siRNA in suppressing pathogenic gene expression [1]. Yet, there are drawbacks that limit siRNA application in vivo: the vulnerability of siRNA against nucleases and the inability of the nucleic acids molecules to diffuse across the cellular membranes [1]. Nanovectors hold great promise for overcoming these limitations towards the development of efficient siRNA therapy. Various nanovectors such as lipids, polymers and inorganic nano-particles have been extensively studied [2]. Among these nanovectors, mesoporous silica nanoparticles (MSN) show promising potential as the next-generation gene delivery carriers [3,4,5]. The main prerequisite for gene therapy is the accumulation of the therapeutic agents on target cancer site [6], which can be achieved through the modification of MSN with targeting moieties such as biological ligand or magnetic particles. Numerous studies on the encapsulation of magnetic nanoparticles (iron oxide) within porous silica for drug delivery have been reported. However, study on MSN with large pores (> 3 nm, termed LPMSN) loaded with magnetic nanoparticles for siRNA delivery is still limited. This work reports the synthesis of novel polyethyleneimine (PEI) functionalized, and iron oxide nanoparticles loaded LPMSN (PEI-Fe-LPMSN) as a multifunctional nano-carrier for gene delivery. LPMSNs with a particle size around 200 nm, a large pore size of 11 nm and a pore connection size of 5 nm are used as a host to impregnate iron oxide nanoparticles inside the mesopores (Figure 1). The large pore in our study has two advantages. Firstly, it provides adequate space for the formation of large iron oxide particles to enhance the magnetic properties. Secondly, the large pore as well as the connection size opens the possibility to adsorb more siRNA molecules not only on the surface but also within the pores, which can enhance siRNA protection against biodegradation. PEI is grafted on the silica pores (PEI-Fe-LPMSN) and surface to enhance the nano-carrier’s affinity against genes and target the cell membrane. Furthermore, polyethylene glycol (PEG) is introduced into the hybrid nanoparticles to improve the biocompatibility and dispersibility. The products have been characterised with various instruments: X-ray photoelectron spectroscopy (XPS), solid-state 13C magic-angle spinning nuclear magnetic resonance (MAS-NMR), Elemental Analysis (EA) and thermogravimetric analysis (TGA). PEI functionalization significantly improves the interaction between the silica particles and oligo DNA / cell membrane, which enhances the cell uptake. The internalization of PEI-Fe-LPMSN can be accelerated by a magnetic field, which causes a high transfection within a short period of time. In addition, the PEGylated nanoparticles show a high biocompatibility. The multifunctional materials enable siRNA delivery within KHOS cancer line and achieve knockdown of PLK-1 oncogenes.

[1] T. Tokatlian and T. Segura, Nanomed. Nanobiotechnol. 2, 305 (2010). [2] H. Shen, T. Sun and M. Ferrari, Cancer Gene Ther. 19, 367 (2012). [3] S.B. Hartono, W. Gu, F. Kleitz, J. Liu, L. He, A.P.J. Middelberg, C. Yu, G.Q. Lu and S. Qiao, ACS Nano,6, 2104 (2012) [4] A.M. Chen, M. Zhang, D. Wei, D. Stueber, O. Taratula, T. Minko, H. He, Small 5, 2673 (2009) [5] S. R. Bhattarai, E. Muthuswamy, A.Wani, M. Brichacek, A. L. Castaneda, S. L. Brock, and D. Oupicky, Pharm. Res., 27, 2556 (2010). [6] S.W. Gersting, U. Schillinger, J. Lausier, P. Nicklaus, C. Rudolph, C. Plank, D. Reinhardt and J.J. Rosenecker, Gene Med., 6, 913 (2004)

NANO-127 Effects of different administration routes on in vivo immune responses for malaria DNA vaccination with magnetic gene vectors F.N. Al-Deen1, C. Ma2, S. D. Xiang3, C. Selomulya1, M. Plebanski3, and R. L. Coppel2 1Department of Chemical Engineering, Monash University, Clayton VIC 3800, Australia 2Department of Microbiology, Monash University, Clayton VIC 3800, Australia 3Department of Immunology, Central Clinical School, Faculty of Medicine, Nursing and Health Sciences, Monash University, Melbourne VIC 3004, Australia

A major drawback of plasmid DNA-mediated gene therapy is low immunogenicity, thus efforts have been put to improve immunogenicity with different enhancing agents. Much attention has been focused on the use of magnetic nanoparticles as carriers for gene therapy. Despite extensive studies on administration routes for naked DNA vaccine, no study has ever directly compared the efficacy and the type of immune response elicited via different routes of administration using superparamagnetic iron oxide nanoparticles (SPIONs) as carriers of DNA vaccine. Here the in vivo delivery of magnetic gene vectors comprising plasmid DNA vaccine encoding Plasmodium yoelii merozoite surface protein MSP119 (VR1020-PyMSP119) condensed on the surface of polyethyleneimine (PEI)-coated SPIONs was investigated via mouse studies in two stages. The first stage determined the optimum N/P ratio (molar ratio of PEI Nitrogen to DNA Phosphate) of magnetic vectors suitable for transfection via a direct intramuscular injection in Balb/c mice. The second stage observed the immune responses generated by these vectors administrated via different routes. Mice were immunised subcutaneously, intradermally, intramuscularly, intraperitoneally, and intravenously on day 0, 14, and 28 with SPIONs/PEI/DNA complexes. The route of administration strongly influenced both total IgG and IgG subclass responses to magnetic vectors containing malaria DNA vaccine. Intraperitoneal immunization route elicited a significantly higher serum antibody titer against PyMSP119 than those elicited by immunization by other methods of injections with the same gene polyplexes. Furthermore, intramuscular immunization led to a slightly higher PyMSP119 antibody responses compared to those obtained by subcutaneous and intradermal injections. Intraperitoneal administration induced robust IgG2a and IgG1 responses respectively, whereas intramuscular, subcutaneous, and intradermal administrations failed to elicit IgG2a and IgG1 responses. Intraperitoneal delivery also allowed repeated injections of high dose of DNA (100 µg) without any sign of toxicity, as opposed to intravenous injection that was proven to be lethal. Vaccination with DNA is often used in conjunction with protein-based subunit vaccines to enhance the effectiveness of DNA vaccine. Hence the immunogenicity of SPIONs/PEI/DNA formulation containing VR1020-PyMSP119 plasmid was also studied using heterologous DNA prime-protein boost vaccination regime for comparison. Three intraperitoneal immunisations with malaria plasmid delivered by magnetic polyplexes, followed by a single protein boost greatly enhanced IgG2a, IgG1, and IgG2b responses respectively, while the intramuscular and intradermal routes generated moderate IgG1and IgG2b responses against PyMSP119 antigen. These outcomes indicated the possibility to design superparamagnetic nanoparticle-based DNA vaccine delivery systems for intraperitoneal delivery that could optimally evoke a desired antibody response for a variety of diseases.

NANO-128 Fundamental design paradigms for systems of interacting nanomagnets D. M. Forrester, E. Kovacs, K.E. Kuerten and F.V. Kusmartsev Department of Physics, School of Science, Loughborough University, Leicestershire, LE11 3TU, UK

As a magnetic field gradient is applied to a magnetic multilayer (MML) of nanodiscs there emerges a series of reversible phase changes (RPC) and/or discontinuous phase changes (DPCs) in the magnetization that are associated with the response of the magnetic moments (MMs) to the field. With understanding of the magnetic field strengths at which these phase changes occur, the precise targeting of the MMLs response characteristics to an applied magnetic field (AMF) can be done. For example, the torque invoked upon a group of nanomagnets by a small AMF may be used in signal transduction therapy, whereby the mechanical stimulation of the cellular membrane of a cancer cell results in a type of cell death [1]. The torque to do this need not be that achieved at the saturation-field and may occur around a DPC of the MM orientation. Thus, we examine the system of interacting MMLs and show the phases associated with magnetization reversals [2]. The movement in and out of cells by particles is also of crucial importance and an interdisciplinary effort is required to understand the mechanisms of transport. Designing the correct structure to attack cancer cells has to be motivated in a large part by finding the optimal geometry for movement of particles through the body. Thus, any design of nanomagnet must be optimised against this prerequisite. In light of this, it is essential for those undertaking this work to be able to understand the response of the nanomagnet to an applied magnetic field because a balance against functionality may have to be met. We investigate the transport and trajectories of nanomagnets that are under the influence of a magnetic field, whilst considering random environmental fluctuations. We also show quasi-static results and in-depth dynamical analyses of interacting clusters of nanomagnets in the AMF [3]. [1] D.-H. Kim, E. A. Rozhkova, I. V. Ulasov, S. D. Bader, T. Rajh, M. S. Lesniak, and V. Novosad, Nature Mater. 9, 165 (2010). [2] D.M. Forrester, K.E. Kuerten, F.V. Kusmartsev, Appl. Phys. Lett. 98, 163113 (2011). [3] D.M. Forrester, E. Kovacs, F.V. Kusmartsev, in preparation (2012)

NANO-129 The design of metamaterials to channel and focus radiation D. M. Forrester and F.V. Kusmartsev Department of Physics, School of Science, Loughborough University, Leicestershire, LE11 3TU, UK

The manipulation of nanoparticles with electromagnetic radiation is an area of intense research and development for medical imaging, controlled drug delivery, bio-molecular sensing, cell apoptosis induction, and creating metamaterials. To understand the difficulties of working with nanoparticles in cancer research, for example, one must understand that they are a hundred to a thousand times smaller than a human cell. Also, there can be ten billion cancer cells in one centimetre squared. One cancer cell is approximately ten micrometers, which is three orders of magnitude larger than a nanoparticle of ten nanometers. The dimensions of nanoparticles are similar to biomolecules: proteins (1-20 nanometers (nm)), DNA (2nm diameter), virus (20nm), cell receptors (10nm), hemoglobin (5nm) and cell membranes (6-10nm). Due to their small size nanoparticles can penetrate a tumour with a high degree of specificity. Thus, one of the biggest issues in designing nanotechnologies is not with the creation of the particles themselves but controlling them when they reach their target. Imaging and diagnostics are thus of paramount importance to create multifunctional therapeutics that can give a patient a quality of life enhancement. Nano devices with dimensions less than 50nm can move freely into cells and organelles, interacting with proteins, enzymes and cell receptors. Even smaller nanoparticles can

travel out of blood vessels and circulate throughout the body (a useful weapon against metastasis). It is clear that as biological processes occur at the nano scale, including events that lead to cancer, nanoparticles could be used to detect disease in a very small volume of cells or tissue. Thus, further understanding of the manipulation of nanoparticles with electromagnetic radiation is of paramount importance. We show our recent results on the control of nanoparticles made from noble and ferromagnetic materials. We introduce artificial pinning points into specially arranged structures of gold nanoparticles and show exceptional electric field enhancements upon irradiation due to a high focusing of plasmons. We integrate ferromagnets with noble metals into heterostructures to give composite qualities and create a material that can both sustain surface plasmons and be magnetically sensitive to light. The modulation of plasmon resonance can be achieved with a magnetic field in these structures. The use of sub-wavelength dimensioned nano-structuring allows these possibilities that lead to ultrafast-magneto-plasmonic switching.We expect that this will ultimately lead to an improvement in live cell analysis using focused radiation. NANO-130 Embedded gold nanoparticle in two dimensional reduced graphene oxide/Porous Silica nanosheet (rGO@PSS-Au) as high sensitive SERS-active nanoprobe for intracellular molecule detecting Y-W Chen, P-J Chen, S-H Hu, S-Y Chen 1Dept. of Materials Sciences and Engineering, National Chiao Tung University 1001 Ta Hsueh Road, Hsinchu, 300 Taiwan, ROC

Surface-enhanced Raman spectroscopy (SERS) is a promising and powerful label free technique for high resolution analysis of molecules. For intracellular analysis, there is a need for SERS-active nanoprobes that are minimally invasive to cells and do not affect cell viability. In this report, we have developed biocompatible, photostable, and multiplexing-compatible surface-enhanced Raman spectroscopic probing material by encapsulating gold nanoparticle in slica porous nanosheet built on reduced graphene oxide (rGO@PSS-Au) for in vitro molecules detecting. rGO@PSS-Au shows linear dependency of Raman signal on different amounts of analytic molecules, CPT, and gives more reproducible and clear signals as compared to normal SERS. These results demonstrate their possibility for the quantification of drug in complex surrounding of living cell. In addition, by depositing different size of gold nanoparticle in suitable porous size of the rGO@PSS, we found that incident light of laser can be trapped for enhancing the Raman signal by many fold as compared to pristine CPT spectra as a reference. Therefore, rGO@PSS-G holds a potential for a direct detection of molecules in living cells with the advantages of easy fabrication, high SERS sensitivity,and biocompatibility. [1] K. K. R. Datta, B. V. Subba Reddy, Katsuhiko Ariga, and Ajayan Vinu, Awgew. Chem. Int. Ed.,49, 5961 (2010) [2] Y. Wang, J. Liu, L Liu, D. D. Sun. Nanoscale Research Letters, 6, 241 (2011) [3] Agnieszka Kaminska, Obianuju Inya-Agha, Robert J. Forster and Tia E. Keyes,Phys. Chem. Chem. Phys., 10, 4172 (2008) [4] Maryuri Roca and Amanda J. Haes, J. AM. CHEM. SOC., 130, 14273 (2008) [5] Elina A. itol, Zul fi ya Orynbayeva, Gary Friedman, Yury Gogotsi, J. Raman Spectrosc. 2012, 43, 817 (2012)

NANO-131 Mucosal immunisation against GnRH-I and GnRH-III conjugates entrapped in lipid nanoparticles A.M. Gebril, E. Bennett, W.H. Stimson, A.B. Mullen, V.A. Ferro University of Strathclyde, Strathclyde Institute of Pharmacy and Biomedical, 161 Cathedral Street, Glasgow, G4 0RE, UK

Lipid based particulates have been studied for their ability to enhance delivery of drugs and vaccines via mucosal and systemic routes [1]. Immunisation of weak immunogenic proteins by mucosal routes requires a suitable delivery system that can enhance immunogenicity. In this study, lipid based formulations were investigated as delivery vehicles for GnRH-based anti-fertility immunogens. Bilosomes [2] were prepared for oral administration, while nasal and vaginal administration was achieved using non-ionic surfactant vesicles (NISV) [3]. These vesicles were loaded with mammalian GnRH-I or lamprey GnRH-III derived sequences conjugated to a range of carrier proteins [4]. The physical characteristics of the vesicles (size, zeta potential, electron microscopy and antigen entrapment efficiency) were characterised. Sexually mature male and female BALB/c mice, were immunised by relevant mucosal (oral, nasal and vaginal) and subcutaneous routes using an established regime [3]. At appropriate time points, specific anti-GnRH antibodies and serum hormone levels were determined. The results obtained indicate that NISV given by subcutaneous and nasal routes enhanced the production of antigen-specific systemic IgG against both GnRH isoforms and caused marked changes in hormone levels. On the other hand, the NISV administered vaginally and bilosomes given orally failed to provoke systemic antibody effects although they produced local sIgA antibodies. We conclude that biodegradable lipid based nanoparticles have strong adjuvants properties when used both nasally or subcutaneously. 1. J.F.S. Mann, et al., Vaccine, 27(27), 3643 (2009). 2. E. Bennett, A.B. Mullen, and V.A. Ferro, Methods, 49(4), 322 (2009). 3. V.A. Ferro, et al., Vaccine, 22(8), 1024 (2004). 4. M.A. Khan, et al., Vaccine, 25(11), 2051 (2007).

NANO-132 Porphyrin-Nanosensors Conjugates:Probing the Sub-Cellular Biochemical Response to ReactiveOxygen Species (ROS). A. S. Lavado1,F. Guntini2, G. Rosser3, A. Beeby3, R. W. Boyle2, W. C. Chan1,and J. W. Aylott1 1University of Nottingham, School of Pharmacy, Nottingham, NG7 2RD,UK 2University of Hull, Department of Chemistry, Hull, HU6 7RX, UK. 3University of Durham, Department of Chemistry, Durham, DH1 3LE, UK.

Reactive Oxygen Species (ROS) are known as important intracellular signalling molecules[1]. These are also well known for their role in oxidative stress and cellular damage, leading to their involvement in several pathologies[2]. Despite the widespread postulation of ROS mechanisms, little is actually known about the immediate response in living cells to the generation of these highly reactive compounds. Tools to quantify and understand ROS production and fate would allow better diagnosis and development of drug targets for ROS related diseases. Our investigation relates to the development of a dual-functionalbiosensing system that would permit the generation of ROS at specific sub-cellular locations and determine in situ the cellular response. In particular, we will target the mitochondria (themain source of ROS generation) and evaluate the mobilisation of calcium as a response to ROS generated. Using a porphyrin-conjugated polyacrylamide nanosensor (~50 nm diameter) we have investigated the nanosensor uptake and sub-cellular localisation in human fetalMesenchymal Stem Cells (hfMSC). Results indicate thatour nanosensors are efficiently internalized andtheylocate to the mitochondria.Ongoing work is focused on the ROS-generating capacity of the nanosensors and how in situ ROS

generation impacts on hfMSC behaviour. We are also studying thecalcium mobilisation response of hfMSC to controlled levels of ROS generated by our dual-functional nanosensors. [1] A. Daiber, BiochimicaetBiophysicaActa 1797, 897 (2010). [2] B. C. Dickinson, C. J. Chang, Nature chemical biology, 7, 8, 504 (2011).

NANO-133 Octa-Ammonium POSS Conjugated Single-walled Carbon Nanotubes (CNTs) as Vehicles for Targeted Delivery of Paclitaxel N. Naderi1, S. Y. Madani1, A. Mosahebi2, A.M. Seifalian1,2* 1UCL Centre for Nanotechnology & Regenerative Medicine, University College London, London, UK 2Royal Free London NHS Foundation Trust, London, UK

CNTs have unique physical and chemical properties [1-4]. Furthermore, novel properties can be developed by attachment or encapsulation of functional groups. These unique properties facilitate the use of CNTs in drug delivery. We developed a new nanomedicine compromised of a nanocarrier, cell-targeting molecule, and chemotherapeutic drug and assessed its efficacy in vitro. Methods: The efficacy of a SWCNT-based nanoconjugate system is assessed in the targeted delivery of paclitaxel to cancer cells. SWCNT’s were oxidized and reacted with octaammonium polyhedral oligomeric silsesquioxanes (Octaammonium-POSS) to render them biocompatible and water dispersable. The functionalised SWCNT’s were loaded with Paclitaxel (PTX), a chemotherapeutic agent toxic to cancer cells and Tn218 antibodies for cancer cell targeting. The nanohybrid composites were characterized with Transmission Electron Microscopy (TEM), Fourier Transform Infrared (FTIR), and UV-vis-NIR. Additionally, their cytotoxic effects on Colon cancer (HT29) and Breast cancer (MCF7) cell lines were assessed in vitro. Results: Increased cell death was observed with PTX-POSS-SWCNT, PTX-POSS-Ab-SWCNT, and free PTX compared to functionalized SWCNT (f-SWCNT), POSS-SWCNT, and cell only controls at 48hr and 72hr time intervals in both cell lines. At all time intervals there was no significant cell death in the POSS-SWCNT samples compared to cell-only controls. Conclusion: The PTX based nanocomposites were shown to be as cytotoxic as free paclitaxel. This important finding indicates successful release of PTX from the nanocomposites and further reiterates the potential of SWCNT’s to deliver drugs directly to targeted cells and tissues. 1. Y. Cui, Q. Wei, H. Park, C.M. Lieber. Science, 293, 5533, 1289-92 (2001). 2. P. Alivisatos, Nat Biotechnol. 22, 1, 47-52 (2004). 3. S. Kim, Y.T. Lim, E.G. Soltesz, et al. Nat Biotechnol, 22, 1, 93-7 (2004). 4. T.A. Taton, , C.A. Mirkin, and R.L. Letsinger. Science, 289, 5485, 1757-60 (2000).

NANO-134 Toxicity study of Fluorescent Nanoparticles for Biological Applications S. Ghaderi1, B. Ramesh1, M. Loizidou1, A. Seifalian1-2

1UCL, Division of Surgery and Interventional Sciences Pond Street, London, UK, NW3 2 QG 2UCL, Centre of Nanotechnology and Regenerative Medicine, Pond Street, London, UK, NW3 2QG

Fluorescent nanocrystals or quantum dots (QDs) are semiconductor nanocrystals (1-20 nm) are emerging as a new class of fluorescent probes for many potential biological and medical applications. Compared to conventional organic fluorescent probes (organic dye), they have substantial advantages such as, bright fluorescence, narrow emission, broad excitation band, high photostability and longer half-life. The overall

objective of this study was to develop an aqueous synthesis method whereby the toxicity was minimized by suitable coatings while maintaining strong emission, stability and biocompatibility for biomedical imaging applications(1). An aqueous one-pot method with a simple approach was undertaken with cadmium telluride (CdTe) QDs. Various processing parameters and coating agents were studied to improve the fabrication and optimize the photoluminescence (PL). In order to reduce or eliminate toxicity of these nanocrystals, a combination of different coating molecules, i.e. mercaposuccinic acid (MSA), Cysteine and mercaptopropyllsobutyl (MPOSS) were investigated. Two cytotoxicity evaluations were carried out using human endothelial (HUVECs) and breast cancer (MCF-7) cells with neutral red staining and fluorescence assay with Alamar blue(2). The presence of functional groups on the surface coatings provided an additional advantage to evaluate conjugation chemistry with biomolecules such as, (antibodies), synthetic peptides (e.g. RGD)(3) and carbon nanotubes (CNTs). The resulting water-soluble QDs exhibited ultra small size (approximately; 5 nm), bright fluorescence and good long-term stability. Thereafter mercury (HgClO4) was introduced to the core of CdTe, which resulted in the near infrared (NIR) scale (800 nm). Introducing MPOSS coatings reduced toxicity significantly even at high concentration to the normal dose. In addition QD successfully was conjugated to CNTs and this was detected under microscopy. Finally, having many advantages, ease of aqueous processing and the excellent PL properties, QDs are widely used in bioapplications as one of most exciting nanobiotechnologies. 1. T. Jamieson, R. Bakhshi, D. Petrova, R. Pocock, M. Imani, A. Seifalian. Biomaterials. 28(31):4717-32; (2007) 2. N. Chen, Y. He, Y. Su, X. Li, Q. Huang, H. Wang. Biomaterials. 33(5):1238-44; (2012) 3. S. Zitzmann, V. Ehemann, M. Schwab. Cancer Res. 62(18):5139-43;(2002)

NANO-135 Influence of the Reaction Conditions in the Preparation of MnO Nanoparticles as T1 Contrast Agents J. Penaranda Avila, M. Colombo, D. Prosperi. Department of Biotechnology and Biosciences, University of Milan – Bicocca. Piazza della Scienza 2. 20126. Milan. Itay.

In recent years, efforts in the development of contrast agents for MRI have been made in order to achieve specific and clearer images and enlargements of detectable organs and systems, which have led to a wide scope of applications of MRI [1]. Some colloidal nanoparticles containing gadolinium (Gd3+) or manganese (Mn2+) have been reported as potent T1 contrast agents due to the fact that they can accelerate longitudinal (T1) relaxation of water protons and exert bright contrast in regions where the complexes localize [2]. MnO nanoparticles dispersed in nonpolar organic solvent were synthesized according to a reported method [1], with some modifications. Varying the reaction temperature (from 280 ºC to 300 ºC), solvent (1-Hexadecene and 1-Octadecene) and reaction time (15, 30 and 60 minutes) we found that the size of the nanoparticles changed significantly and reproducibly as well as their morphology (spherical, cubic and star-like). Characterization of the nanoparticles was carried out using different techniques such as dynamic light scattering (DLS), transmission electron microscopy (TEM), thermo gravimetric analysis (TGA). Nanoparticles phase transfer was possible throughout their encapsulation in an amphiphilic polymer (poly(isobutylene-alt-maleic anhydride) PMA) and the measurements of the ion concentration of the solutions were done by ICP-AES. The transversal relaxiviy (r1) of these water-dispersed nanoparticles was measured and it was found that they change for each type of sample as well as the ratio r1/ r2, which vary from 1.4 to 2.1 consistent with that is usually reported in the literature [1], [2]. Finally, surface functionalization, as well as the influence of these morphological differences between nanoparticles in their biocompatibility, cellular uptake and interactions with biomolecules is going to be the next step of this work. [1] H.B. Na, J.H. Lee, K. An. et al. Angew. Chem., 119, 5493, (2007). [2] C.-C. Huang, N.-H. Khu and C.-S. Yeh. Biomaterials., 31, 4073, (2010).

NANO-136 CYTOTOXIC EFFECT OF PHOTOEXCITED TITANIUM DIOXIDE-DNA NANOCOMPOSITES ON THE NASAL CANCER CELLS IN VITRO R.R.Bazak1,2, M.S. Elwany2, S.A. Tawfik,2, T. Paunesku1, G.E. Woloschak1 1Northwestern University, Chicago, USA 2University of Alexandria, Egypt

Malignant cells frequently contain mutations in growth promoting proto-oncogenes such as c-Myc which has been found to be activated in many tumors.(1-7) Manipulation of c-Myc expression has been shown to induce apoptosis in several studies.(8-10) This puts emphasis on the concept of oncogene addiction which is consistent with the notion that malignant cells are often "addicted to" the continued activity of specific overexpressed oncogenes for maintenance of their malignant phenotype(1) and that reversal of an oncogene can inhibit cancer cell growth and could potentially be translated to improved survival rates.(1, 11) Hybrid titanium dioxide-DNA nanocomposites can specifically target a certain gene(12) whilst retaining the photocatalytic properties inherent to titanium dioxide,(13). Therefore, photoexcited c-Myc targeted titanium dioxide nanocomposites are expected to induce apoptosis in malignant cells expressing c-Myc oncogene. The study was conducted on Roswell Park Memorial Institute 2650 cell line (RPMI 2650). The expression of c-Myc in this cell line was verified by conducting a western blot test. Core-shell 2 nanoparticles made of iron oxide core and a shell of titanium dioxide were synthesized and tagged with Alizarin red stain. The nanoparticles were characterized by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), atomic force microscopy, visual spectral analysis and dynamic light scattering. A segment of the DNA complementary to c-Myc was extracted from plasmid containing c-Myc gene was obtained from GeneCopoeia (Catalog No. EX-Z2845-B09) and attached to the core-shell nanoparticles via a dopamine linker. Intranuclear localization of the nanocomposites was achieved by coating them with a nuclear localization peptide (KH peptide). The hybridization ability of the core-shell nanocomposites was verified by FISH. The experiment was designed such that the cells were either incubated with no nanoparticles, bare nanoparticles (without peptide or DNA), nanoparticles coated with KH peptide or nanocomposites coated with both nuclear localization peptide and c-Myc complementary DNA. Each group of cells was either exposed to ultraviolet radiation for five minutes, 2Gy ionizing radiation or kept in the dark at room temperature for an equivalent duration. The cells were fixed after 30 minutes and stained for 53BP1 and after 24 hours and stained for M30 CytoDEATH. Incubation of cells with KH-nanocomposites followed by exposure to ultraviolet light resulted in a decrease in cell volume, chromatin condensation and nuclear fragmentation as well as a diffuse cytoplasmic positivity for M30 CytoDEATH indicating apoptosis.(14) When these cells were stained for 53BP1 they appeared smaller in size compared to controls and some of them had a characteristic deeply stained pyknotic nucleus, shrunken cytoplasm with no nuclear staining for 53BP1. In contrast, the control cells that were incubated with KH-nanocomposites but not exposed to ultraviolet light did not show any morphologic feature of apoptosis nor did they stain for M30 CytoDEATH. This suggests that KH-nanocomposites are able to induce apoptosis upon excitation with ultraviolet light. The cells that were incubated with KH-DNA nanocomposites and then exposed to ultraviolet light demonstrated more pronounced features of apoptosis compared to all other treatment groups. These cells exhibited marked decrease in cell volume, chromatin condensation, nuclear fragmentation and a cytoplasmic speckled rather than diffuse positivity for M30 3 CytoDEATH. When stained with 53BP1, these cells were smaller in size and more pleomorphic compared to their controls. Two subgroups of cells were observed; some cells had more than one DNA double strand break while others demonstrated a characteristic deeply stained pyknotic nucleus and a shrunken cytoplasm with no nuclear staining for 53BP1. The observed DNA double strand breaks in the first subgroup of the cells but not in controls is an indirect evidence of photo-induced endonuclease activity of titanium dioxide nanocomposites described by Paunesku et al.(12) The subgroup of cells exhibiting deeply stained nuclei, shrunken cytoplasm and no positivity for 53BP1 observed among those incubated with KH-DNA nanocomposites were most abundant compared to other treatment groups. The present study shows that c-Myc targeted photoexcited DNA-titanium dioxide nanocomposites are

able to initiate significant apoptosis in RPMI 2560 in vitro. These results suggest that DNA-titanium dioxide nanocomposites could be adopted as a novel anti-cancer modality for tumors in the future. [1] I.B. Weinstein, Science., 5, 63 (2002) [2] C.S. Fan, N Wong, S.F. Leung, et al., Hum Pathol., 31, 169 (2000) [3] J. Blancato, B. Singh, A .Liu, et al., Br J Cancer., 19, 1612 (2004) [4] B. Nakata, N. Onoda, Y.S . Chung, et al., Gan To Kagaku Ryoho., 22, 176 (1995) [5] D .Hawksworth, L . Ravindranath, Y. Chen, et al., Prostate Cancer Prostatic Dis., 13, 311 (2010) [6] C.F. Rochlitz, R. Herrmann, E. de Kant., Oncology., 53, 448 (1996) [7] C.H. Chen, J. Shen, W.J. Lee, et al., Int J Gynecol Cancer.,15, 878 (2005) [8] D.W. Felsher, J.M. Bishop. Mol Cell., 4, 199 (1999)

NANO-137 The potential application of functionalized ZnO nanorod as electrochemical (glucose and metal ions) biosensors for Intracellular environment. Muhammad H. Asif1 , Rizwan Raza1 and Magnus Willander2

1Department of Physics, COMSATS Institute of Information Technology, Lahore-54000. Pakistan. 2Department of Science and Technology (ITN), Linköping University, SE-60174, Sweden

The nanostructure of zinc oxide (ZnO) such as nanorods and nanowires has interesting nanosurfaces in addition to its bulk properties. Recently ZnO have attracted much interest because of its unique piezoelectric, semiconducting, catalytic properties and being biosafe and biocompatible morphology combined with the easiness of growth. This implies that ZnO has a wide range of applications in optoelectronics, sensors, transducers, energy conversion and medical sciences. This abstract relates specifically to electrochemical (glucose and metal ions) biosensors for intracellular environment based on functionalized zinc oxide nanorods for biochemical applications. To adjust the sensor for intracellular measurements, the ZnO nanorods were grown on the tip of a borosilicate glass capillary (0.7µm in diameter) and functionalized with polymeric membrane or enzymes for intracellular selective metal ion or glucose sensors. The sensor in this study was used to detect and monitor real changes of metal ions and glucose across human fat cells and frog cells using changes in the electrochemical potential at the interface in the intracellular microenvironment. The fabrication of such type of device aims to explain the methodology of ions sensing using functionalized ZnO nanorods for intracellular environment. This nanoelectrode device paves the way to enable analytical measurements in single living cells. 1 M. H. Asif, A. Fulati, O. Nur, M. Willander, Cecilia Bränmark, Peter Strålfors Sara I Börjesson and Fredrik Elinder., “Functionalized

zinc oxide nanorod with ionophore-membrane coating as an intracellular Ca2+ selective sensor”, Applied Physics letters. 95 (2). 2009.

2 M. H. Asif, Syed M. Usman Ali, O. Nur, M. Willander, Cecilia Brännmark, Peter Strålfors, Ulrika Englund , Fredrik Elinder and B. Danielsson, Functionalized ZnO nanorod based intracellular glucose sensor. Biosensors and Bioelectronics 25. 2010. 2205-2211.

3 M. H. Asif, Syed M. Usman Ali, O. Nur, M. Willander, Ulrika.H Englund , Fredrik Elinder.Functionalized ZnO nanorod-based selective magnesium ion sensor for intracellular measurements. Biosensors and Bioelectronics 26. 2010.

4 M. H. Asif, O. Nur, M. Willander, and B. Danielsson., “Selective Calcium Ion Detection with Functionalized ZnO Nanorods-Extended Gated MOSFET”, Biosensors and Bioelectronics 24 2009.

NANO-138 Effect of the diamond nanopowder on bacterial strains: Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. K. Adach1, J. Skolimowski2, P.Ceynowa3, K. Mitura1,3

1 Technical University of Lodz , Institute of Material Science and Engineering, Lodz,1/15 Stefanowskiego st., Poland 2 University of Lodz, Department of Organic Chemistry, Lodz, 12 Tamka st., Poland. 3Koszalin University of Technology, Institute of Mechatronics, Nanotechnology and Vacuum Technology, 75-453, Poland. Presenting author: Katarzyna Mitura Koszalin University of Technology, Institute of Mechatronics, Nanotechnology and Vacuum Technology, 75-453, Poland.

Diamond nanopowders due to its properties may form ideal biomaterial for biomedical applications. The surface of nanodiamond can be modified in many different ways [1, 2, 3]. Due to their surface ability to connection with many of organic groups, we can attached not only the organic groups but also antibiotics [3]. Nanodiamonds have a developed surface area, which can be modified using chemical reactions [4]. The aim of made reactions, was obtain into the surface of nanodiamond various kind of "linkers" [4]. Reactions was carried out in order to form the alkyl chains and aromatic rings on the nanodiamond surface. It is possible to carry out such chemical synthesis in order to bond an antibiotic in a large distance from the nanodiamond surface. Such modifications carry out to increase the number of antibiotic molecules on the nanodiamond surface. This study was confirmed by biological tests on bacterial strains: Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739, Candida albicans ATCC 10231. This study shows that nanodiamonds induce a low fluorescence in bacteria and thus the calcium stress. Powders with a solution of bacteria and fungi, showed respectively green and blue fluorescence under UV light. This property of the nanodiamonds can suggest it as diagnostic microbiological medium. [1] S. Rojas, J. D. Gispert, R. Martin, S. Abad, C. Menchon, D. Pareto, V. M. Victor, M. Alvaro, H. Garcia, J. R. Herance, Biodistribution of Amino-Functionalized Diamond Nanoparticles. In Vivo Studies Based on 18F Radionuclide Emission, ACS Nano, vol. 5, no. 7, 5552–5559, (2011). [2] Solarska K., Gajewska A., Skolimowski J., Woś R., Bartosz G., Mitura K., Effect of non-modified anod modified nanodiamond particles by Fenton re action on human endothelial cells, Journal of Achievements In Materials and Manufacturing Engineering, Vol. 43, 2, 2010, 603 - 607 [3] Adach K., Skolimowski J., Mitura K., Chemical modification of nanodiamond particles produced by detonation method, Elektronika, 11, 2011, 84 – 86 [4] A. Krueger, The structure and reactivity of nanoscale diamond, J. Mater. Chem., 18, 1485–1492, (2008)

NANO-139 Development of polymeric nanocapsules containing clobetasol propionate by in situ polymerization for topical use S. P. Carneiro1, J. M. C. Vilela2, O D. H. Santos1 1Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brasil. 2Fundação Centro Tecnológico de Minas Gerais – Belo Horizonte, Minas Gerais - Brasil

Clobetasol propionate is a very potent topical corticosteroid commonly prescribed for treatment of some skin disorders, like psoriasis, atopic and contact dermatitis, due to its vasoconstrictive, anti-inflammatory and anti-proliferative effects [1]. However, it has been reported that the long-term used of clobetasol propionate is related to adverse effects, such as skin atrophy, contact allergies and even systemic effects. Besides that, its use in conventional formulations is associated with low patient compliance to treatment, because of their sensory greasy and difficulties in applying in large areas [2]. In order to overcome these drawbacks, there were

developed polymeric nanocapsules from nanoemulsions obtained by PIT method. Clobetasol propionate nanoemulsions containing acrylate derivate monomers were developed by Phase Inverse Temperature (PIT), a low-energy emulsification method. From these systems, polymeric nanocapsules were produced by in situ polymerization, which started after the addition of FeSO4/KPS solutions and all process took around 4 hours [3]. There were obtained 4 different formulations: one nanoemulsion using 2-Ethylhexyl Acrylate monomer, which led to the nanocapsule formed by the poly(2-Ethylhexyl Acrylate) and another nanoemulsion using a blend of 2-Ethylhexyl Acrylate and Buthyl Acrylate monomers which gave rise to the nanocapsule with the copolymer poly(2-Ethylhexyl Acrylate-Buthyl Acrylate). All systems were well characterized by size measurements and polydispersity, showing similar values between themselves. Atomic Force Microscopy was also employed for surface and morphology particle analyses. Finally, drug release tests were performed with Franz type diffusion cells to assess the amount of clobetasol propionate released in function of time. [1] S. Wiedersberg, C. S. Leopold and R. H. Guy. EurJPharmBiopharm, 68, 3, 453, 2008. [2] N. Lowe, S. R. Feldman, D. Sherer, J. Weiss, J. S. Shavin, Y. L. Lin, V. Foley, P. Soto. JDermTreat, 16, 3, 158, 2005. [3] P. L.Goto, J. M. C. Vilela, M. S. Andrade., O. D. H. Santos. JDispSciTech. 2012.

NANO-140 NOVEL METHODS FOR DETECTION OF INTRAVENOUSLY ADMINISTERED TITANIUM DIOXIDE NANOPARTICLES IN TISSUES OF TRANSGENIC MICE S.N.El Achy1,2, N.M. Eldeeb2, M.A. Hamza2 T. Paunesku1, G.E. Woloschak1, 1Northwestern University, Chicago, 2University of Alexandria, Egypt Visualization of nanoparticles without inherent optical fluorescence properties such as titanium dioxide posed a significant problem to nanoscience research involving cells and animal tissues. Bulk forms of titanium dioxide are generally biologically and chemically inert; however, the surface of titanium dioxide nanoparticles smaller than 20 nm is reactive because of ‘corner defects’ that are acquired as a novel property by virtue of their nanoscale size. These corner defects readily bind with ortho-substituted bidentate ligands such as dopamine and Alizarin Red S (ARS) making a very attractive mechanism for functionalization.(1-4) In light of this, titanium dioxide nanoparticles can be tailored to serve as multimodal imaging agents as it can be conjugated to optically fluorescent agents as Alizarin Red S, optical dyes, as well as contrast agents for magnetic resonance imaging (MRI) such as iron oxide and gadolinium.(5-7) Furthermore these nanoparticles, when linked to DNA oligonucleotides, have a potential to cleave intracellular genomic DNA in a sequence specific and inducible manner and therefore can be utilized as gene knockout devices and as possible tumor imaging agents.(1, 3, 8) Despite the extensive research into the use of TiO2 nanoparticles, the lack of variety in detection techniques has hindered statistical analysis and large-scale monitoring of organ and 2

NANO-141 Serum Adsorption Affects Cellular Uptake Profile and Consequent Impact of Nanoparticles on Human Lung Cell Lines M. Lim1*, R.Tedja1, R. Amal1, C. Marquis2

1School of Chemical Engineering, The University of New South Wales, Sydney NSW 2052, Australia. 2 School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney NSW 2052, Australia.

The effects of nanoparticles exposure to serum protein is an important consideration in the design of nanoparticles for bio-imaging and drug delivery [1]. It is shown herein that serum protein adsorption reduces the aggregate size of titania nanoparticles, affecting their uptake into A549 and H1299 human lung cell lines. Initially, the cellular uptake of the serum-treated titania was lower than that of untreated titania. Expulsion of particles was then observed, followed by a second phase of uptake of serum-treated titania. This resulted in a large increase in the cellular content of serum-treated titania. Surface adsorbed vitronectin and the clathrin-mediated endocytosis pathway were shown to regulate the uptake of titania into A549 cells, while the endocytosis mechanism responsible remains elusive for H1299. Intriguingly, nystatin treatment was shown to have the unexpected effect of increasing nanoparticle uptake into the A549 cells via an alternate endocytic pathway. The surface adsorbed serum components were also found to provide some protection against the cytotoxic effect of endocytosed nanoparticles. [1] Roslyn Tedja, May Lim, Rose Amal, and Christopher Marquis, ACS Nano, Vol. 6, No. 5, 4083–4093 (2012). NANO-142 A Numerical Model to Reduce Guesswork in the Design of Nanoparticles for Nanomedicine M. Lim1*, H.Wiogo1, R. Amal1

1School of Chemical Engineering, The University of New South Wales, Sydney NSW 2052, Australia.

A numerical model that can predict the size stability of nanoparticles in biological media is shown herein. The model can be used as the basis for designing stable polymer functionalised magnetic nanoparticles for application in biomedical imaging and drug delivery. The model takes into account key factors that affect the stability of magnetic nanoparticles in biological media, including the volume fraction of aggregates, absolute zeta potential, thickness of adsorbed steric layer, and conformation of the steric layer on the surface of nanoparticles. The model shows a polyionic (polyethylenimine or polymethacrylic acid) coating can easily stabilize the nanoparticles in water. However, unless the polymer can attract a layer of serum protein onto the nanoparticles surface to produce additional steric hindrance, this size stability will be lost when the nanoparticle is transferred into biological media. Size of the nanoparticles is also shown to be an important factor. Larger magnetic particles (>30 nm) with strong magnetisation (~85 emu/g) will always possess a deep secondary minimum due to long-range magnetic force. The short-range electrostatic and steric repulsive forces, caused by the presence of polyionic and/or protein coatings, could not diminish this secondary minimum, unless a steric layer of over 100 nm in thickness was attained. The developed model agrees well with the experimental observations, and reduces the guesswork that is often involved in the design of nanoparticles system for application in nanomedicine.

NANO-143 Durable, Washable, Photosterilisable Textiles with Binder-free Nano Titania S. A.M. Tofail1, P. Cronin1, E. Dworniczek2, J. Zeglinski1, I. Buzalewicz3, R. Franiczek2, M. Wawrzyńska2, Halina Podbielska3, Peter Tiernan4

1 Department of Physics and Energy and Materials and Surface Science Institute, University of Limerick, Limerick, Ireland 2Department of Microbiology, Medical University of Wroclaw, Wroclaw, Poland 3Institute of Biomedical Engineering and Instrumentation, Wroclaw University of Technology, Wroclaw, Poland 4Department of Design & Manufacturing Technology, University of Limerick, Limerick, Ireland

At any time, over 1.4 million people worldwide suffer from infectious complications acquired in hospital. Infections acquired in health care settings are among the major causes of death and increased morbidity among hospitalized patients. Photocatalytic nanoparticles are effective against hospital bugs, such as in killing MRSA. We have used these nanoparticles to develop antimicrobial textiles that can be photosterilised. The patent pending technology binds nanoparticles to textiles without using any chemical binder. This binding is durable and retains antibacterial action even after 40 domestic wash cycles. When exposed to UV radiation from sunlight or other sources, textiles with such nanoparticles are spontaneously photosterilised. A casual walk in the sun for a few minutes thus cleanses the wearer’s apron or drape. Indoor lights will have similar results but photosterilisation will take slightly longer time. The BioElectricSurface research (www.bioelectricsurface.eu) leading to these results has received funding from the European Community's Seventh Framework Programme (FP7/2007-2011) under grant agreement n°212533. This communication reflects the views only of the authors, and the Commission cannot be held responsible for any use which may be made of the information contained therein. NANO-144 Microwave Sensing of Common Enzymes with Nanocrystalline Hydroxyapatite O. Korostynska1, A. A. Gandhi2, M. Salazar-Alvarez3, A. Mason1, A. Al-Shamma'a1, E. Magner3, S. A.M. Tofail2

1School of Built Environment, BEST Research Institute, Liverpool John Moores University, Liverpool, UK 2Department of Physics and Energy and Materials and Surface Science Institute, University of Limerick, Ireland 1Department of Chemical and Environmental Scienxe and Materials and Surface Science Institute, University of Limerick, Ireland.

Hydroxyapatite (HA) is a leading biocompatible material extensively used as grafts and grafts, coatings and scaffolds. Recently reported high piezo and pyroelectricity of nanocrystalline HA [1], its polarisation by electron beam [2] and selective adsorption of proteins on polarised domains [3] indicate the potential biosensing applications of nanocrystalline hydroxyapatite. For this purpose, a comprehensive understanding of the dielectric behaviour of different forms and stoichiometry of HA over a range of frequency bandwidth relevant for biomedical sensing is critical. Such information for HA, especially its frequency dependent dielectric behaviour over the GHz range is relatively rare. To this end, we present a systematic investigation of dielectric properties of HA in the GHz frequency range. We will also demonstrate how the behaviour of HA at the GHz range can be utilised in developing a new generation of highly biocompatible bio-sensors based on HA for quantitative detection of common enzymes such as cytochrome C, ferritin and biotin. [1] Lang, S.B. et al. App. Phys. Letters, 2011. [2] Plecenik, T. et al., Appl. Phys. Letters, 2011 [3] Robin, S. et al., Langmuir 2011

NANO-145 Electrostatic Nanodomains on Biomaterials: A Novel Methodology to Study Biological Interactions with Surface Charge S.A.M. Tofail1

1Department of Physics and Energy and Materials and Surface Science Institute, University of Limerick, Ireland

It is commonly understood that electrical properties such as local electrostatic charge distribution at biomaterials’ surfaces play a significant role in defining biological interactions. Biomaterials that are currently used in such devices, e.g. in cardiovascular and urinary stents and coatings in hip prosthesis, do not specifically address this interfacial phenomenon in device designs. A detailed knowledge of such interactions at the nanometre scale will not only produce a selective biological response but also pre-screen many inappropriate designs of biomedical devices long before any expensive animal or potentially risky clinical trials. Here we will present some of the key methodologies of electrical modifications of biomaterials’ surfaces and the relevance of such modifications in our understanding of biological interactions with surface charge in biomaterials used in therapeutic applications such as in cardiovascular stents, urological stents and orthopaedic implants and grafts. The focus of the presentation will be on nanoscale confinement of electrostatic charge to create electrostatic nanodomains with a minimal to a zero level of alteration in surface chemistry and morphology. This restriction on unmodified surface chemistry and morphology has been imposed to strip out the real and actual role of surface charge in mediating biological reactions on biomedical device surfaces [1, 2]. [1] www.bioelectricsurface.com [2] S.A.M. Tofail (ed.), Biological Interactions with Surface Charge in Biomaterials (Royal Society of Chemistry: Cambridge, 2011).

NANO-146 Mechanical and tribological properties of nanocomposite carbona-C:H/Ti silver doped layers for medical applications L. Kolodziejczyk, W. Szymanski, D. Batory Lodz University of Technology, Institute ofMaterials Science and Engineering, Stefanowskiego 1/15, 90-924 Lodz, Poland

Due to favorable mechanical, tribological and biomedical properties the carbon layers are of interest of many branches of the industry [1]. Growing interest in Ag doped DLC layers is observed over the last few years. Both, well known antibacterial properties of silver [2] as well as good biocompatibility of carbon layers [3] constitute the outstanding solution for a variety of applications, especially for medical implants. The aim of this study is to evaluatethe influence of silver on mechanical and tribological properties of nanocomposite DLC coatings. Carbon doped silver coatings were producedusinga hybridRFPACVD/MS and consist of two stages: synthesis of nanocompositecarbon doped titanium coatings [4] followed by carbon and silver deposition onto carbon coating. Carbon layers synthesis was performed with use of the classic RF PACVD process in methane atmosphere whereas as the source for titanium ions the pulsed magnetron sputtering (MS) process was applied. Second stage was performed in the same reaction chamber but the PVD process was carried out using the silver cathode. Due to the application of the gradient of chemical composition Ti – C it is possible to manufacture thick and well adherent carbon layers with very good mechanical, tribological and corrosion parameters. Application of silver as the doping material allowed the modification of the mechanical and biological properties of manufactured layers as a function of silver amount (C:Ag ratio). [1] C. Donnet, Surf Coat Tech, 100-101, 180 (1998). [2] J. S. Kim et al., Nanomed Nanotech Biol Med, 3, 95 (2007)

[3] S.E. Rodil,R. Olivares, H. Arzate, S. Muhla, Diam Relat Mater, 12, 931 (2003) [4] M. Clapa and D. Batory, J. Achiev Mater Manuf Eng, 1-2, 415 (2007)

NANO-147 Erosion of human dental enamel after exposure to teeth storage liquids and acidic drinks investigated by AFM, CLSM and NI techniques. W. Szymanski1, L. Kolodziejczyk1, A. Ostrowska2

1Lodz University of Technology, Institute of Materials Science and Engineering, Stefanowskiego 1/15, 90-924 Lodz, Poland 2Medical University of Lodz, Institute of Dentistry, Pomorska 251, 92-213 Lodz, Poland

Dental erosion is usually defined as an irreversible loss of dental hard tissue due to a chemical process, without the involvement of bacteria. Various surveys have shown that there is a high prevalence of this teeth disorder among patients [1]. What is more, it has been widely acknowledged that a predominantly acidic diet is one of the most important induced factors [2,3]. A wide range of techniques has been used to recognize even the earliest signs of this process during in vitro studies [4]. The authors want to investigate the matter with the highest precision, taking into account even the smallest changes into mechanical or morphological properties of enamel. It is of crucial importance that the first stage of dental erosion, called the initial demineralization, starts when a tooth surface is exposed to acidic food or beverage. It has been claimed that the most modern techniques related to in vitro dental erosion research are AFM, CLSM and nanoindentation [5]. In case of such sensitive techniques, we have to take into consideration all the possible factors which may have an influence on the results of our research. One of the factors which has been too often neglected is the condition of the teeth samples storage [6]. The aim of this study is to analyze the influence of teeth sample storage on modern techniques of dental erosion measurements, comparison of three methods of the erosion evaluation and determine the erosive potential of Powerade and orange juice. [1] H. El Aidi, E.M. Bronkhorst and G.J. Truin, J Dent Res 87, 731 (2008) [2] A. Lussi, T. Jaeggi and D. Zero, Caries Res, 38, 34 (2004) [3] V. Sirimaharaj, L. Brearley Messer and M.V. Morgan, Aust Dent J, 47(3), 228 (2002) [4] T. Attin, Dental Erosion. From Diagnosis to Therapy, 20, 216, G. M. Withford, Karger, Basel Switzerland (2006) [5] Finke M et al., Surf Sci, 491, 456 (2001) [6] S. Habelitz et al., J Biomech, 35(7), 995 (2002)

NANO-148 Fluorescent Nanosensors for Measuring the Cellular Environment J.W. Aylott1

1School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK Nanoscience and nanotechnology have opened new avenues for analysis of biological systems. One manifestation of this has been the reduction in size of the measurement tool, resulting in nanoscale sensing devices. Fluorescent nanosensors are an example of these new measurement tools and in this presentation the synthesis, characterisation and application of fluorescent nanosensors sensitive to a variety of analytes including pH, oxygen and proteases will be discussed. These examples demonstrate the versatility of the nanosensor architecture in enabling a range of chemistries to be used for specific conjugation and modification purposes to produce nanosensors with enhanced performance characteristics. The nanosensors described will be based on polyacrylamide and silica sol-gel matrices, ranging in size from 30-300 nm, and containing combinations of

fluorescent dyes to produce ratiometric sensing nanodevices. The presentation will also detail the application of these sensors in a range of biological systems. These will include: Mapping the pH of the intestinal tract of C.elegans, a non-parasitic soil nematode which has traditionally been used to model complex biological processes such as aging, genetics, neuroscience, and drug response. Measuring cellular health and metabolism in 3D cell cultures grown on electrospun polymer scaffolds. Intracellular delivery and measurement in a range of cultured cell lines. Fluorescent nanosensors have great potential for measuring the cellular environment with exquisite spatial and temporal resolution, however, there are a number of challenges to overcome before the technology matures and is accepted into the mainstream of bioanalysis and a critical assessment of these will be presented. NANO-149 Specific targeting by (oligo)mannose functionalized hydroxyethyl starch nanocapsules: en route to drug delivery systems with targeting properties H. Freichels1, M. Wagner1, P. Okwieka2, R.G. Meyer2, K. Landfester1, A. Musyanovych1*, V. Mailänder1, 2

1Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany 2III. Medical Clinic (Hematology, Oncology and Pulmonology), Langebeckstr. 1, 55131 Mainz, Germany

Hydroxyethyl starch nanocapsules (NCs) are interesting hydrophilic drug delivery carriers, since they do not show unspecific interactions with the living cells [1]. Only the presence of a targeting agent on their surface allows them to target specifically the desired site of action. In this paper, we report the synthesis and cell uptake of crosslinked hydroxyethyl starch (HES) NCs decorated with (oligo)mannose, which is an effective targeting agent for macrophage and dendritic cells [2]. The crosslinked HES NCs were prepared via interfacial polyaddition of HES with 2,4-toluene diisocyanate (TDI) in inverse (water-in-oil) miniemulsion [3] and then functionalized with (oligo)mannose following two different strategies. To compare the activity and availability of a targeting agent, different types of mannose molecules such as α-D-mannopyranosylphenyl isothiocyanate, 3-O-(α-D-mannopyranosyl)-D-mannose and α3,α6-mannotriose were used for functionalization of NCs. The availability of the mannose was unambiguously assessed by interaction with a fluorescent lectin. Moreover, the accessibility of the pilot molecule was improved by the presence of a PEG linker at the surface of the NCs. To simulate in vivo conditions, where proteins interact with nanoparticles with possible hindrance of the accessibility to the targeting agent, the mannosylated NCs were first incubated with human serum before interaction with the fluorescent lectin. Enhancement of uptake into dendritic cells demonstrates the targeting function in in vitro studies. [1] Baier, G., Baumann, D., Siebert, J.-M., Musyanovych, A., Mailänder, V., Landfester, K. Biomacromolecules, accepted (DOI: 10.1021/bm300653v) (2012) [2] Guermonprez, P., Valladeau, J., Zitvogel, L., Thery, C., and Amigorena, SAnnu. Rev. Immunol. 20, 621 (2002) [3] Landfester, K., Musyanovych, A., and Mailander, V. J. Polym. Sci. Pol. Chem. 48, 493 (2010)

NANO-150 A Continuous Counterflow Nanoconcentrating Module for Online Monitoring in Closed Bioreactors Haakon Karlsen, Tao Dong*, Xinyan Zhao Norwegian Centre of Expertise on Micro and Nanotechnology, Department of Micro and Nano Systems Technology –IMST, Vestfold University College,

This study introduces the counter-flow nanoconcentrator (nC) as a non-clogging continuous online sampling module for concentration monitoring in closed fermentor bioreactors and/or pharmaceutical plants without contamination. Nanofluidic characteristics were thoroughly investigated for continuous counter-flow nano concentration. The modern pharmaceutical factories often use bioreactors to produce drugs. The bioreactor is required to be a closed system with a specific environment inside. It is also required to monitor the concentration of drug molecules inside the mixture in bioreactors. The sampling device is the critical component to not only take the sample outside the close system but also keep the bioreactor free of contamination. We have developed a semipermeable sampling module which will only allow the biological macromolecules to gate through. The structure is non-clogging and very stable, which could be integrated in the online monitoring system of the bioreactor. The two main filtration principles of filtration are dead-end (direct/normal flow filtration) and crossflow filtration (tangential flow filtration)[1]. Due to particle adhesion on the membrane of dead-end filters, clogging is inevitable. Due to the tangential flow for crossflow filter, the adhered particles are removed by the flow shear forces, but the crossflow filter has reduced particle separation efficiency. The nC is a derivation of the microconcentrator (µC) [2, 3] which employs the counter-flow principle in a concentrator unit with velocity optimized elliptical design, consisting of a solid front part and a barrier back part with a filtrate hole in the middle. The barrier consists of optimized aerofoil shaped micropillars (with the purpose to guide the flow) positioned in an elliptical shape where adjacent pillars forms variating cross-section penetrating channels between them with a minimum gap size of 40 nm. The entrance to the penetrating channel causes gradual decrease in local velocity while the narrow part of the gap causes a sharp increase in velocity. When the main flow containing particles passes around the counter-flow unit, the bigger particles tend to be entrained by the main flow and pushed away from the channel between adjacent pillars. The aerofoil shape is designed so that the smaller particles can remain near the pillars when travelling past a counter-flow unit, and is sucked in due to the increased velocity in the gap [2]. This counter-flow design reduces risk of clogging-failure. The height of the structures is limited by the aspect ratio of the Deep Ion Reactive Etching (DRIE) process. With a gap size of 40 nm and an aspect ratio of perhaps 30-50, a structure height of 1.2-2 µm is obtainable. In this study, the authors present simulations demonstrating the counter-flow principle, with focus on the biological sampling module application. Nanofluidic investigations are conducted through FEM simulation of single penetrating channels and full units by using laminar flow module in COMSOL 4.2 and Navier-Stokes (NS) equations. According to Travis et al. [4] classical NS-equations can be used for channel widths bigger than 10 molecular diameters (for water about 2.5 nm), 40nm>>2.5nm, NS-equations are therefore assumed safe. Estimations of Knudsen number for water also places the flow in the continuum regime. To obtain pressure and velocity profile along the penetrating channel, a cut line is taken through the middle of the gap. The main flow velocity used is kept at 1cm/s which is in the range of common nanofluidic device velocities; 1 cm/s - 1 µm/s [5]. The penetrating channel simulation is performed to investigate velocity and pressure, while full unit simulation is performed to investigate efficiency. The simulated results show the expected behaviour. The velocity has a gradual decrease into the channel, as well as a sharp increase in the gap. The full unit simulation shows that there is flow in all channels. Due to the effective length of the 40 nm gap being very short, there is a very big fluidic resistance and the differentiated pressure in the gap is very high. This allows the nC unit to be used as a semipermeable sampling module in a closed system as the high fluidic resistance can serve as an analogy to the input impedance of an operational amplifier, where the inputs draws little current. [1] R. Levy and M. Jornitz, Advances in Biochemical Engineering/Biotechnology, vol. 98, pp. 1-26 (2006)

[2] T. Dong et al. Microfluidics and Nanofluidics, vol. 10, pp. 855-865 (2011). [3] N. Tran-Minh, T. Dong, Q. Su, Z. Yang, H. Jakobsen, and F. Karlsen, Biomedical Microdevices, vol. 13, pp. 179-190 (2011). [4] K. P. Travis, B. D. Todd, and D. J. Evans, Physical Review E, vol. 55, pp. 4288-4295 (1997). [5] T. M. Squires and S. R. Quake, Reviews of Modern Physics, vol. 77, pp. 977-1026 (2005).

NANO-151 HIV envelope detection using Whispering Gallery Mode Sensing A. Ma,2, Y-J Chen3, F.Vollmer3, B.L.J. Webb1, K. Wilson4, D. Holmes2 Department of Infection1 & London Centre for Nanotechnology2, UCL, UK. Max-Planck Institute3, Erlangen, Germany. Department of Chemistry4, Imperial College London, UK.

Whispering gallery mode (WGM) biosensing is a surface-based sensing technique. Monitoring microresonator optical resonances enables molecular binding data. In addition to it being real-time, label-free, and high throughput, the strong light-matter interactions makes it a promising candidate for ultrasensitive measurements over existing techniques such as SPR. Much progress has been made in realising its potential in single virus and particle detection [1] as well as in engineering integrated devices [2] for point-of-care applications. Here we present a novel method to detect HIV envelope (gp120) proteins using WGM resonator-immobilised, high-affinity, anti-HIV envelope antibody fragments (VHH), derived from llamas [3]. These possess higher VHH affinity for envelope than any conventional antibody or CD4 (the physiological HIV receptor) itself. We demonstrate a technology for HIV POC testing by integrating VHH with sensitive binding detection in a WGM biosensor. By monitoring different WGM resonances, we propose a method towards distinguishing between envelope and virus, which SPR cannot achieve. [1] Vollmer F, Arnold S, Keng D. Single virus detection from the reactive shift of a whispering-gallery mode. Proceedings of the National Academy of Sciences. 105(52), 20701–20704 (2008). [2] Harazim SM, Bolanos Quinones VA, Kiravittaya S, Sanchez S, Schmidt O. Lab-in-a-Tube: on-chip integration of glass optofluidic ring resonators for label-free sensing applications. Lab Chip. (2012). [3] McCoy LE, Quigley AF, Strokappe NM, Bulmer-Thomas B, Seaman MS, Mortier D, Rutten L, Chander N, Edwards CJ, Ketteler R, Davis D, Verrips T, Weiss RA. J Exp Med. 2012 Jun 4;209(6):1091-103.

NANO-152 The Effect of N-Terminal Substitution on the Hydrogelation and Biocompatibility of Dipeptide Hydrogelators. K.W.J.Currie, R.F. Houston, D.J. Adams, S.P. Gorman, B. Walker, B.F. Gilmore School of Pharmacy, Queens University Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast BT9 7BL, UK

Peptide hydrogelators have attracted significant research interest due to their inherent biocompatibility and potential applications in the areas of tissue engineering and controlled drug delivery. An interesting class of hydrogelators are those that can ‘self-assemble’ from simple dipeptide conjugates [1]. These short peptides spontaneously self-assemble in an aqueous environment through non-covalent interactions such as hydrogen bonds and π-π stacking interactions to generate complex fibrous networks, which lead to immobilising of the liquid phase. The most extensively characterised of these are dipeptides N-terminally conjugated to either Fmoc or naphthalene. [2-3]. Here we report on the synthesis and characterisation of novel N-terminally substituted diphenylalanine dipeptides. We examined the effects of varying the N-terminus on the ability of these compounds to act as hydrogelators and their resulting physico-chemical and mechanical qualities. A range of experimental techniques was used to evaluate each compound’s gelation properties. The mechanical properties formed gels was analysed using oscillatory rheology. Fourier transform infra-red spectroscopy (FTIR) was used to

1

assess the secondary structure of self-assembled gels. Transmission electron microscopy (TEM) was used to examine the microstructure and assess fibre dimensions. The biocompatibility of these dipeptides was also evaluated in vitro for potential tissue engineering or drug delivery applications. We show that conjugation of benzoic acid (BA) at the dipeptide N-terminus promotes the self-assembly and hydrogelation. In addition to this, we demonstrate that fluorinationof this N terminal moiety (4FBA) improves the viscoelastic properties of the formed hydrogels, compared to Fmoc-FF (Figure 1) and was found to be nontoxic towards the human keratinocyte cell line HaCaT. This study demonstrates the importance of N-terminal substitution in the rational design of low molecular weight peptide hydrogelators. NANO-153 Shear stress sensitive nanocontainers with tunable release properties M.N. Holme1,2,3, T. Saxer,1, A. Zumbuehl2, B. Müller3

1Cardiology Division, University Hospitals of Geneva, 1211 Geneva, CH 2Department of Chemistry, University of Fribourg, 1700 Fribourg, CH 3Biomaterials Science Center, Basel University, c/o University Hospital, 4031 Basel, CH

There are several pathologies that lead to a change in endogenous shear stresses within the vascular tree, and therefore raise the possibility of treatment by targeted drug delivery using a purely physical trigger. In one example, atherosclerosis, plaque deposits constrict the coronary artery, leading to an increase in the endogenous wall shear stresses by a factor of at least ten. We recently reported the design, synthesis and formulation of lenticular vesicles formed from an artificial 1,3-diamidophospholipid, Pad-PC-Pad. In a proof-of-principal study, these vesicles were passed once through a diseased or healthy plastic model artery at physiological pressure and flow rate in isotonic solution, and preferentially released their contents in the diseased model [1]. Here, we present findings from an expanded 100 nm unilamellar vesicle library formulated from several artificial phospholipids synthesised in our lab. Increasing the hydrogen bonding in the lipid bilayer by introduction of a urea instead of amide linker led to an increase in stability to high shear stresses (>20 Pa). Addition of two carbons to each chain of this urea analogue decreased shear sensitivity further, with no preferential release observed. This was corroborated in shear stress experiments by shaking samples on a vortex for 60 seconds. Additional hydrogen bonding and longer chain length led to much longer release profiles, more suitable to single-injection, long circulating applications. One-to-one mixtures of phospholipids with different tail lengths were found to be intrinsically more stable than vesicles of either individual component or other ratio mixtures. Such findings are an important step towards understanding how phospholipid formulations might be tuned to release their contents at a pre-defined shear stress, and could find applications in targeted delivery for many diseases, such as atherosclerosis, stroke, intestinal ischemia and renal infarction. [1] M.N. Holme, et al, Nature Nanotech., 7, 536 (2012)

NANO-154 Study on the Interaction between Peptide Functionalised Colloidal Quantum Dots (CdSe/ZnS) and DNA Strands K. Pechstedt1, T. Melvin1

1Optoelectronics Research Centre, University of Southampton, Highfield, Hampshire, SO17 1BJ, U.K.

There exists indirect experimental evidence that short oligonucleotides and double-stranded calf thymus DNA strands can non-specifically adsorb onto the surface of colloidal quantum dots (QDs).[1-4] Recent experimental

polymerase chain reaction (PCR) studies in the presence of QDs have also implied selective binding of QDs with single stranded and double stranded DNA.[5-7]. Here, the association of peptide-functionalised QDs (CdSe/ZnS) with either double stranded or single stranded lambda-phage DNA was investigated. For the case of single stranded DNA evidence for the association of QDs to DNA was observed for strands stretched under sheer flow conditions. The emission properties of the structures have been investigated and it is suggested that QDs are assembled in clusters along the strand and that the ends of the strand wrap around single QDs. The suggested QD-DNA binding mechanism is a hydrogen bonding mechanism between protonated carboxyl groups of the peptides on the QD surface and the DNA nucleotide bases. Furthermore, fluorescent structures of the same type were not observed when the QDs are incubated with double stranded lambda-phage DNA, suggesting the increased flexibility of single stranded compared to double stranded DNA facilitates DNA-QD association. These results will be discussed. Cellular titanium dioxide nanoparticle interaction.(1) Herein we describe new techniques that have been developed for detection of Fe3O4@TiO2 core-shell nanoparticles in tissues of transgenic mice with prostate cancer, using Alizarin Red S for optical fluorescent imaging and Dopamine-Streptavidin-Chromogen staining for optical light microscopy in comparison to the gold-standard technique X-Ray Fluorescence microscopy previously described by Paunesku et al.(3, 9) Glucose coated Fe3O4@TiO2 core-shell nanoparticles were injected intravenously in seven male transgenic mice prostate cancer models that are known to develop adenocarcinomas similar to the poorly differentiated variants of human prostate cancer at seven months of age.(10) The control group of mice comprised five transgenic mice with prostate cancer that were not injected with nanoparticles and one mouse without prostate cancer and not injected with nanoparticles. One week after the procedure, the mice were sacked and their internal organs (liver, spleen, heart, lung, brain and prostate) were dissected, fixed in formalin and submitted for histopathological examination. Alizarin Red S post-staining of the tissues was performed for demonstration of tissue distribution of nanoparticles using fluorescence microscopy at a wavelength of 510nm (TissueGnostics Cell Analyzer-Vienna). A newly developed technique involving Biotin-Streptavidin complex that was later incubated with chromogen dye was explored for detection of titanium dioxide nanoparticles using standard optical microscopy at magnifications reaching 1000X (Axioscope-Zeiss). The tissues were also processed and examined using the XFM facility at the 2ID-E and 2ID-D beamlines at the Advanced Photon Source at Argonne National Laboratory which is best equipped for mapping and quantifying elements between Si and Zn in the periodic table. Examination of the mouse tissues using the XFM determined the uptake of Fe3O4@TiO2 core-shell nanoparticles by the prostatic carcinoma tissues in addition to cells of the reticuloendothelial system within the liver and spleen. 3 A good correlation existed between the positive signals in the XFM maps with the Alizarin Red S staining using the fluorescent microscope as well as the Biotin-Streptavidin staining using the optical light microscope. No positive signals were detected in the mouse control tissues that were not injected with nanoparticles for both techniques. This confirms that the newly developed techniques may be further utilized for detection of TiO2 in body tissues without the need for expensive sophisticated microscopes. Additionally, all three methods used for detection have verified that intravenously administered glucose coated Fe3O4@TiO2 core-shell nanoparticles accumulate in the lung alveoli and bronchial epithelium; the macrophages in the splenic red pulp; the Kupffer cells in the liver sinusoids as well as the renal tubular epithelium. New methods for staining and detection of Fe3O4@TiO2 core-shell nanoparticles within tissues have been developed using Alizarin Red S staining for detection using a fluorescent microscope as well as Biotin-Streptavidin-Chromogen staining for detection using a standard optical microscope. This will open new frontiers in nanoscience research allowing scientists to monitor the nanoparticle-organ and tissue interactions at larger scales, for better control of targeted nanotherapeutic devices. [1] K.T. Thurn, T. Paunesku, A. Wu, E.M. Brown, B. Lai, S. Vogt, et al., Small., 5, 1318 (2009) [2] K.T. Thurn, H. Arora, T. Paunesku, A. Wu, E.M. Brown, C. Doty, et al., Nanomedicine., 7, 123 (2011) [3] T. Paunesku, T. Rajh, G. Wiederrecht, J. Maser, S. Vogt, N. Stojicevic, et al., Nat Mater., 2, 343 (2003) [4] H.P. Wu, T.L. Cheng, W.L. Tseng., Langmuir., 23, 7880 (2007) [5] T. Paunesku, T. Ke, R. Dharmakumar, N. Mascheri, A. Wu, B. Lai, et al., Int Rev Cytol., 4, 201 (2008) 4 [6] P.J. Endres, T. Paunesku, S. Vogt, T.J. Meade and G.E. Woloschak., J Am Chem Soc., 129, 15760 (2007) [7] J. Kurepa, T. Paunesku, S. Vogt, H.Arora, B.M. Rabatic, J. Lu, et al., Nano Lett., 10, 2296 (2010) 8. E.M.B. Brown, A. Wua, K.T. Thurn, B. Haley, J. Clark, T. Priester, G.E. Woloschak. Anal Biochem., 383, 226, (2008)

9. T. Paunesku, S. Vogt, B. Lai, J. Maser, N. Stojicevic, K.T. Thurn, et al. Nano Lett., 7, 596 (2007) 10. I.G. Maroulakou, M. Anver, L. Garrett, J.E. Green. Proc Natl Acad Sci U S A., 8, 11236 (1994)

NANO-155 Polymeric nanoparticle development for targeting and modulating macrophage functions E. Almouazen, S. Bourgeois, H. Fessi, S. Briancon University Lyon 1, LAGEP UMR-CNRS 5007, F-69622, Villeurbanne, France

In malignant tumors, the infiltration of macrophages was well documented and the so-called tumor associated macrophages (TAM) are thought to support both tumor progression and metastatic invasion. The aim of this study was to target TAM by polymeric nanoparticles (NP) in order to modulate their functions or benefit from their infiltration. In this goal, two kinds of active were encapsulated. The first is all-trans retinoic acid (RA) an immunomodulator with modest stability and pharmacokinetics. The second is the active metabolite of vitamin D3 (25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3) which are potential anti-cancer agents but with high risk of hypercalcemia. The intratumoral injection of NP ensures passive targeting of TAM that specifies the activity of loaded drug. Firstly, the internalisation of NP by TAM was studied in vitro and in vivo. Fluorescent microscopy observations demonstrated that Nile Red labelled NP were highly engulfed and sustained over seven days inside bone marrow derived macrophages in vitro. The intra-tumoral injection of NP into xenograft glioma nude mice confirmed that NP were efficiently uptaken by TAM. Secondly, RA was encapsulated with high encapsulation efficiency (90%), a sustained release over 96h and enhanced stability. RA loaded NP induce modulation of gene expression on macrophages as evaluated by RT²-PCR array [2]. Finally, the active metabolites of vitamin D3 were encapsulated to localize their action on tumor. An in vitro study on a cell line of breast cancer (MCF-7) showed the advantage of NP to extend and enhance their antiproliferative activity [3]. We can conclude that using NP offers a great possibility to target TAM. This targeting is useful for modulating macrophages functions or for using macrophages as storage of anti-cancer agent. [1] F. Chellat, Y. Merhi, A. Moreau, L.Yahia, Biomaterials, 25, 7260 (2005). [2] E. Almouazen, S. Bourgeois, A. Boussaïd, P. Valot et al., Int. J. Pharm., 430, 207 (2012). [3] E. Almouazen, S. Bourgeois, L. Petter Jordheim, H. Fessi, S. Briançon, submitted to Pharm.Res (2012).

NANO-156 Monitoring the organization of multicellular tissue constructs by magnetic manipulation and dual imaging R. Di Corato1, F. Gazeau1, C. Le Visage2, G. Autret3, P. Mowat4, D. Letourneur2, O. Tillement4, C. Wilhelm1

1Laboratoire MSC, Université Paris Diderot, CNRS UMR 7057, Paris 75205, France. 2INSERM, UMR 698, Paris 7 - Denis Diderot, Paris 13 – Nord, Paris 75877, France. 3INSERM, U970, Paris Cardiovascular Research Center – PARCC, Paris 75015, France. 4Laboratoire PCML, Université Lyon 1, CNRS UMR 5620, Villeurbanne 69622, France.

In regenerative medicine, new methods in which transplanted tissues are grafted with different cells and new techniques to image them non-invasively have become highly desirable. Up to date, MRI cell imaging is one of the best candidate to track transplanted cells distribution, by using mainly T2 contrast agents as superparamagnetic nanoparticles (SPIO).[1] However, only one cell type can be detected on MR images. Here we propose a new tissue engineering imaging approach taking advantage of a dual-cell labelling with respectively T1 and T2 agents, to monitor two independent cell types. In details, mesenchymal stem cells (MSCs) were incubated with gadolinium oxide nanoparticles.[2] For the first time it was possible to detect cells as white spots at a cellular resolution in scaffolds. To this novel cellular T1 imaging was associated a standard T2 imaging, by labelling

endothelial HUVEC-C cells with 7 nm-SPIO. Different constructs were tested, using planar or tubular scaffold for cells seeding based on biodegradable polysaccharide gels. All the scaffolds were analyzed by a 4.7T MRI, equipped with a CryoProbe (Bruker). By applying T1-weighted RARE or 3D-FISP-FID sequences, a signal from each cell type was acquired at the same time. Beyond this proof of dual imaging, the magnetic responsivity of labelled HUVEC-C cells was profited to design functional scaffolds. In particular the blood vessel engineering is an example of multicellular patterning, in which HUVEC-C were magnetically-driven to coat exclusively the lumen surface, whereas the MSC homed in the gel core. The outlined geometry was successfully imaged by MRI, by collecting simultaneously a darkening in presence of endothelial cells and a bright signal from the scaffold media, as confirmed by confocal microscopy analysis. [1] C. Wilhelm et al., Biomaterials, 29, 3161 (2008) [2] J.L. Bridot et al., J. Am. Chem. Soc., 129, 5076 (2007).

NANO-157 Designing a Glucose Biosensor Using Electrospun Carbon Nanofibers (CNF) H. F. Huq1,J. Acuña2, M. Braden3, E. M. Piñon3, 1University of Texas-Pan American, Edinburg, Texas, 78539, USA. 2Pharr-San Juan-Alamo Independent School District, Texas, USA. 3McAllen Independent School District, Texas, USA

A bienzymatic glucose biosensor that uses vertically aligned carbon nanofibers (VACNF) has been shown to detect glucose [1,2]. However, manufacturing VACNF is an expensive process. The goal of this research is to determine if non-aligned, thus less costly, electrospun carbon nanofibers (ECNF) incorporated as part of a biosensor are also be able to detect glucose. Second, Tests were performed using a manual three-electrode analyzer, with positive results at glucose concentrations as low as 2 mM. This research consists of two parts – 1) To study the characteristics of ECNF in various solutions and 2) To develop a prototype of ECNF based glucose biosensor. All five dry ECNF samples are found to be ohmic and exhibit linear behavior within the range -2.226 V to 2.992 V and the operating frequency is within the range of 13 Hz to 850 kHz. Loose ECNF are added to an aqueous bienzyme solution to evaluate its effectiveness as part of a glucose detector. As the ECNF samples’ length increase and their cross-sectional area remain constant, it is predicted that their resistance increases and thus lower their output current. Data beyond this range behaved in a nonlinear manner. In addition, this study shows that ECNF does allow glucose to be detected. Its lower cost makes it a more economical choice over VACNF. Carbon nanofiber (ECNF) is one of the potential candidates for biosensors, specifically to detect glucose, an important biosignatures to diagnose diabetes. The use of this type of biosensor in other medical applications such as in detecting other maladies would greatly benefit the general population. [1] I. Willner and B. Willner, Nano Lett., 10, 3805 (2010). [2] A. B. Islam et al., IEEE Sensors Journal, 11, 2798 (2011)

NANO-158 Mesoporous Silica Contrast Agents for the Endoscopic Detection of Nascent Colorectal Cancer N.T. Chen1, 2, 3, J.S. Souris3, V. Konda4, U. Dougherty4, M. Bissonnette4, C.Y. Mou2, *, L.W. Lo1, *, and C.T. Chen3, *

1Division of Medical Engineering Research, National Health Research Institutes, Zhunan Miaoli 350 (Taiwan)

2Department of Chemistry, National Taiwan University, Taipei 106 (Taiwan) 3Department of Radiology, The University of Chicago, Chicago IL (USA) 4Department of Medicine, The University of Chicago, Chicago IL (USA).

Cell surface glycosylation is known to play a number of significant and different roles in the modulation of physiologic and pathophysiologic pathways in human biology. In cancer, for example, deviations in the composition/structure and areal density/distribution of glycosylation-associated molecules on cell surfaces are common occurrences in tumorigenesis and neoplastic progression – strongly correlating with tumor invasiveness, metastatic potential, and the evasion of host immuno-surveillance[1,2]. To determine the utility of targeting aberrant glycosylation in the detection of nascent colorectal cancers in situ, we have recently begun to design, synthesize, and evaluate dysplastic adenoma-targeting mesoporous silica nanoparticles (MSNs) for use as endoscopic contrast agents. MSNs, approximately 100 nm in diameter, were synthesized via conventional sol-gel chemistry, with the incorporation of fluorescein isothiocyanate (FITC) within the MSN’s silica framework during co-condensation – to protect the fluorophore from photobleaching and O2 quenching, and to maximize the nanoparticle’s available surface area for targeting ligand conjugation. Following nanoparticle synthesis, the exteriors of MSNs were labeled with the lectin Ulex europaeus agglutinin 1 (UEA-1 -L-fucose that is known to be expressed on the luminal surface glycoproteins of colorectal dysplastic adenomas and nascent cancers[3-5]. Morphologies of the resulting functionalized MSNs were then characterized by transmission electron microscopy, dynamic light scattering, N2 adsorption–desorption isotherm analysis, and zeta potential measurement. In vitro studies of MSN- -L-fucose positive Caco-2

-L-fucose negative HCT116 human colorectal cancer cells. In vivo endoscopic exams were performed using a commercially available clinical colonoscope and probe-based confocal fluorescence microscope, with fasted/prepped A/J male mice that had been subjected to azoxymethane (AOM) / dextran sodium sulfate (DSS) treatments – a standard model for colitis-associated colorectal tumor development. In vitro and endoscopic images, topical delivery of MSN-UEA-1 to human colorectal adenocarcinoma cells and in situ murine AOM/DSS-induced colorectal polyps/tumors bot -L-fucose. Using non-fluorescent UEA-1 pretreatment confirmed UEA- -L-fucose binding specificity. Ongoing studies are aimed at enhancing our nanoplatform’s transport through colon mucus and its dwell-time once pathology bound. [1] J.M. Rhodes, R.R. Black, A. Savage, J Clin Pathol, 39, 1331 (1986) [2] F. Gabor, E. Bogner, A. Weissenboeck, M. Wirth, Advance Drug Delivery Review, 56, 459 (2004) [3] S. Yuan, C.A. Roney, J. Wierwille, C.W. Chen, B. Xu, J. Jiang, H. Ma, A. Cable, R. Summers, Y. Chen, Phys. Med. Biol. 55(1), 191 (2010) [4] C.A. Roney, J. Xie, B. Xu, P. Jabour, G. Griffiths, R.M. Summers, Proc. of SPIE, 6916, 691610 (2008) [5] X. Wang, I. Kochetkova, A. Haddad, T. Hoyt, D.M. Hone, D.W. Pascual, Vaccine, 23, 3836 (2005)

NANO-159 Nanocapsules generated out of a polymeric dexamethasone shell suppress the inflammatory response of liver macrophages M. Fichter1, G. Baier2, L. Pretsch1, K. Landfester2 & S. Gehring1

1Children’s Hospital, Johannes-Gutenberg University, Langenbeckstr. 1, 55131 Mainz, Germany 2Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany

Dexamethasone (DXM) is a synthetic glucocorticoid with anti-inflammatory and immunosuppressive properties. Targeted delivery of dexamethasone to key cells regulating inflammation, such as Macrophages, Monocytes, and Kupffer cells represents a promising approach to minimize side effects. Therefore, aim of the present study was to evaluate the ability of two different DXM-nanocapsule formulations to suppress the inflammatory response of phagocytic cells. The two formulations are characterised by their nanocapsule (NC) structure with one formulation having a hydroxyethylated glucose polymer (HES) shell encapsulating DXM and the second nanocapsule consisting exclusively out of a DXM shell. All types of nanocapsules were synthesized by the inverse (water-in-oil) miniemulsion process through an interfacial polyaddition reaction resulting in an average size of 240 nm (DXM) and 215 nm (HES-DXM), respectively. Non-parenchymal murine liver cells, with an approximate 30 % Kupffer cells, served as target cells. HES-DXM nanocapsules were incorporated by F4/80 positive Kupffer cells and CD31 positive endothelial cells. In comparison, DXM-NCs were taken up to a greater extent and predominately by endothelial cells. The release of the NC-content was confirmed by incorporation of CellTracker™ into the NCs and the subsequent fluorescent staining of nanocapsule positive cells documented by confocal microscopy. Up-take of DXM capsules significantly reduced the release of inflammatory cytokines (IL-6, TNF-α) in particular by LPS stimulated Kupffer cells. In conclusion, biodegradable DXM nanocapsules are efficiently incorporated by phagocytic cells, such as Kupffer cells, and subsequently suppress the release of inflammatory cytokines in response to LPS stimulation. Importantly, the DXM nanocapsules, consisting exclusively out of a dexamethasone shell, bear the potential to serve as a carrier for additional therapeutics. NANO-160 Plasma synthesis of biocompatile carbon- hydroxyapatite nanostructures A. Niedzielska, P. Skwierczyński, W. Jakubowski, P. Komorowski, W. Mróz, D. Batory Lodz University of Technology, 90-924 Lodz, 1/15 Stefanowskiego St. Poland

Both carbon and HAp (hydroxyapatite) coatings found wide application spectra in medicine [1,2] as modifications improving biological properties of medical implants. Authors of this work developed a method of synthesis of carbon-hydroxyapatite nanostructures combining properties of those materials. This report presents the results of preliminary examinations related to the synthesis and characterization of mechanical and biological properties of composite carbon-HAp nanostructures deposited onto Ti6Al7Nb titanium alloy pre-modified with use of nanocrystalline diamond intelayer. Characterization of the mechanical features, performed by means of nanoindentation method revealed in dependence on the process parametecrs nanohardness in the range between 4 – 6,5 GPa and adhesion between 6 – 21 mN. Further physicochemical features were examined with use of SEM and FTIR. Obtained structures exhibit hydrophilic character. As a biological material were selected: cell line of human osteoblast Saos-2 (ATCC, Manassas, USA), Endothelial cells line EA.hy 926 (ATCC, Manassas, USA) and E. coli bacteria. Mammalian cells and bacteria were cultured in standard conditions. The culture was carried out for 48 hours in the case of eukaryotic cells, and bacterial culture was carried out for 24 hours. For the evaluation of proliferation, cytotoxicity and susceptibility to bacterial biofilm formation marking method a "live / dead" based on the fluorescent techniques was used. Tested surfaces in different ways affected the proliferation

of mammalian cells but none of these surfaces showed cytotoxicity against endothelial cells and osteoblasts. Surfaces of carbon – HAp nanostructures showed high resistance to microbial colonization and formation of bacterial biofilm. 1. W. Mróz, A. Bombalska, B. Budner, S. Burdyńska, M. Jedyński, A. Prokopiuk, E. Menaszek, A. Ścisłowska-Czarnecka, A.

Niedzielska, K. Niedzielski: Comperative study of hydroksyapatite and ostacalcium phospate coatings deposited on metallic implants by PLD method" Appl Phys. A (2010) 101 713-716

2. D. Batory, J. Gawroński, W. Kaczorowski, A. Niedzielska, C – HAp composite layers deposited onto AISI 316 L austenitic steel, Surface & Coatings Technology 206 (2012) 2110–2114

NANO-161 Three dimensional interpenetrating network nano-porous sacnffold on titanium substrate and its cellular biocompatibility Chengyun Ning1, Yongmei Ge1, Guoxin Tan2, Caixia Xu1, Huade Zheng1, Liming Fang1, Jingwen Liao1, Peng Yu

1 College of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 PR China. 2 Institute of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510640 PR China.

A novel three-dimensional nano-network porous film on titanium (Ti) surface was fabricated by an alkali treatment method. Surface characterizations of the network layer were carried out by scanning electron microscopy (SEM), X-ray diffraction (XRD), energy disperse spectra（ EDS） , and contact angle and surface tension analysis. Human mesenchymal stem cells (hMSCs) were seeded on the nano-network porous film and cultured for 7 days in vitro. Cell attachment and proliferation on the film were evaluated by SEM and MTT assay. The results showed that the film morphology and structure depended on the length of alkali treatment time. After alkali treatment for 48 h, three dimensional nano-network porous film formed on the Ti surface . There was no clear interface between the film and the substrate. The nano-network porous surface significantly reduced water contact angle compared with that prior to alkali treatment. Moreover, alkali treated Ti surface dramatically enhanced hMSCs attachment and proliferation than untreated titanium surface. Taken together, the above results demonstrate that the nano-network porous film on Ti surface renders improved biocompatibility [1] T. Hryniewicz, R. Rokicki, K. Rokosz. Surf. Coat. Technol. 203, 1508 (2009). [2] 3. R. Singh, P.D. Lee, Trevor C, et al. Acta Materialia. 5, 477 (2009). [3] M. Balazic, J. Kopac, J. M. Jackson, and W. Ahmed. Int. J. Nano Biomater. 1, 3 (2007).

NANO-162 PVP-directed synthesis of Nano- Hydroxyapatite under a new growth mechanism Chengyun Ning1, Zhaoyi Yin1,2, Guoxin Tan3, Huade Zheng1, Jingwen Liao1, Peng Yu1

1 College of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 PR China. 2 College of Materials Science and Engineering, Kunmin University of Science and Technology, Guangzhou 510641 PR China. 3 Institute of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510640 PR China.

Hydroxyapatite (HAP) is the principal inorganic constituent ofbones and teeth. Nano-hydroxyapatite (HAP) has received a greatdeal of attention due to its high surface interaction and bioactive properties. Long rod-like, short rod-like and spherical nano-hydroxyapatite(HAP) with different sizes has been prepared using polyvinylpyrrolidone(PVP) as a soft-template, and PVP plays adecisivepart in controlling the morphology and particle size. The experimental results showed thatthecrystallinity of HAP decreased with the increase ofthe content of PVP, and astrong chemical bondingexistedbetweenthe PVP moleculesand HAP.Based on the

experimental results, a new growth mechanismof nano-hydroxyapatite wasproposed.PVPaffects the four steps of HAP formation respectively, i.e.,nucleation, surface-regulating, growth and oriented attachment, which depends oncoordinative bonding, steric effect and hydrogen bond. [1] J.D. Chen,Y.J. Wang, K.Wei, S.H.Zhang, X.T.Shi, Biomaterials 28, 2275 (2007). [2] Y.J. Zhu, X.L. Hu, W.W.Wang. Nanotechnology 17, 645 (2006).

NANO-163 In vivo biodistribution of POSS-nanocomposite polymer coated quantum dots using Inductively Coupled Plasma Mass Spectroscopy and optical studies S.B. Rizvi1, S.Y. Yang1, A. Darbyshire1, S. Taniguchi4, M. Green4, A.M Seifalian1,2,3* and M. R. Keshtgar1,3.

University College London (1Centre for Nanotechnology & Regenerative Medicine, UCL Division of Surgery & Interventional Science, 2London Centre of Nanotechnology), London, United Kingdom,3Royal Free Hampstead NHS Trust Hospital, London, United Kingdom. 4Department of Physics, King’s College London

We have previously reported the synthesis and characterization of a type II core/shell/shell near infrared (NIR) emitting semiconductor quantum dot (QD) based on CdTe/CdSe/ZnSe[1]. Near Infra Red emitting QDs can be visualised in deep tissues as the biological window is transparent to these wavelengths (650-900 nm)[2]. The main limitation to their in vivo application is their toxicity as most QDs are based on chalcogenide salts of Cadmium, Tellurium and Selenium. We have also synthesized a novel nanocomposite polymer based on Polyhedral Oligomeric Silsesquioxane - Poly(carbonate-urea) Urethane (POSS-PCU) and used it for various tissue engineering applications including vascular grafts and recently tracheal graft and nasolacrimal duct that has successfully been implanted into humans.[3, 4] We used the emulsion form of this polymer to coat the QDs in order to take them to a clinical setting. In this paper we examined the effect of two different QD surface coatings on their in vivo biodistribution. The coating materials used included 1) Mercaptoundecanoic acid (MUA) and 2) POSS-PCU nanocomposite emulsion. POSS-PCU was synthesised by a process of emulsion polymerization and optimised to encapsulate CdTe/CdSe/ZnSe QDs for colloidal stability, biocompatibility and enhanced photostability. Male Sprague Dawley rats were injected intravenously with a fixed dose (2 nmoles) of POSS-PCU coated (cQDs) and MUA coated QDs (uQDs) for a period of 1 and 24h (n=4/ group). Organs were harvested for histology and QD biodistribution was quantified by determination of elemental Cd concentration using Inductively Coupled Plasma Mass Spectroscopy (ICPMS) and fluorescence optical imaging. Both POSS-PCU coated (cQDs) and MUA coated (uQDs) QDs localised maximally to the liver at 1 and 24h. The cQDs had a longer systemic circulation half-life and significantly enhanced in vivo photostability at 24h compared to the uQDs. The cQDs were found to be excreted by the hepatobiliary route as compared to uQDs which were excreted by the kidneys due to their small size. Both QDs injected into the hind leg of a rat model localised to the inguinal lymph node and were tracked using a live NIR Imaging system. Histopathology showed no evidence of organ toxicity for cQDs compared to uQDs. The enhanced photostability of the cQDs led to a significantly higher fluorescence signal in all organs at 24h which did not correlate with Cd concentrations on ICPMS results. We concluded that the POSS-PCU coating can significantly enhance in vivo stability, biocompatibity and circulation half-life of NIR QDs for a range of in vivo imaging applications particularly sentinel lymph node localisation in cancer surgery. Also, evaluation of QD biodistribution by fluorescence optical imaging is a semi-quantitative method and may be biased by the enhanced photostability imparted by different surface coatings of QDs. Hence, ICPMS remains the gold standard for a quantitative assessment of QD biodistribution. In addition, POSS-PCU polymer has already been used in humans as surgical implants for trachea, bypass graft and a nasolacrimal duct conduit, hence making the regulatory tasks for clinical application of polymer coated QDs much easier. [1] S.Taniguchi, M.Green, S.B.Rizvi, A.Seifalian, J. Mater. Chem. 2011, 21, 2877. [2] H.Kobayashi, S.Kawamoto, M.W.Brechbiel, M.Bernardo, N.Sato, T.A.Waldmann, Y.Tagaya, P.L.Choyke, Neoplasia. 2005, 7, 984.

[3] K.Chaloupka, M.Motwani, A.M.Seifalian, Biotechnol. Appl. Biochem. 2011, 58, 363. [4] P.Jungebluth, E.Alici, S.Baiguera, B.K.Le, P.Blomberg, B.Bozoky, C.Crowley, O.Einarsson, K.H.Grinnemo, T.Gudbjartsson, G.S.Le, G.Henriksson, O.Hermanson, J.E.Juto, B.Leidner, T.Lilja, J.Liska, T.Luedde, V.Lundin, G.Moll, B.Nilsson, C.Roderburg, S.Stromblad, T.Sutlu, A.I.Teixeira, E.Watz, A.Seifalian, P.Macchiarini, Lancet 2011, 378, 1997.

NANO-164 A novel POSS-PCU nanocomposite polymer as a biocompatible coating for quantum dots S.B. Rizvi1, S.Y. Yang1, A. Darbyshire1, S. Taniguchi4, M. Green4, M. R. Keshtgar1,3 and A.M Seifalian1,2,3*

University College London (1Centre for Nanotechnology & Regenerative Medicine, UCL Division of Surgery & Interventional Science, 2London Centre of Nanotechnology), London, United Kingdom,3Royal Free Hampstead NHS Trust Hospital, London, United Kingdom. 4Department of Physics, King’s College London

Quantum dots (QDs) are fluorescent nanoparticles with unique photophysical properties that enable them to potentially replace traditional organic dyes and fluorescent proteins in various bio-imaging applications. However, the inherent toxicity of their cores based on cadmium salts, limits their widespread biomedical use. We have developed a novel nanocomposite polymer emulsion based on Polyhedral Oligomeric Silsesquioxane Poly (carbonate-urea) Urethane (POSS-PCU) that can be used for various biomedical applications. We have previously used the solid form of this polymer for various tissue engineering applications including vascular grafts, nerve grafts and more recently as a tracheal graft and nasolacrimal duct that has successfully been implanted into humans[1-4]. Here we report the synthesis and characterization of the emulsion form of this novel POSS-PCU nanocomposite polymer and describe its application for coating QDs for biological application. The polymer was synthesized by a process of emulsion polymerization and formed stable micelles of ~ 33nm in diameter. Core/shell/shell QDs based on CdTe/CdS/ZnS and CdTe/CdSe/ZnSe were efficiently stabilised by the polymer emulsion through encapsulation within the polymer micelles. Characterisation studies showed no significant change in the unique photophysical properties of QDs after coating. The polymer was biocompatible to HepG2, HUVECs and Mouse skeletal muscle cells at 2.5% after a 24 hr exposure on in vitro testing. Polymer encapsulated QDs showed enhanced photostability on exposure to high degrees of UV irradiation and air as well as significantly reduced cytotoxicity on exposure to Hep G2 cells at 30µg/ml for 24h. We have therefore concluded that the POSS-PCU polymer emulsion has a potential to make a biocompatible and photostable coating for QDs enabling a host of biomedical applications to take this technology to the next level. [1] T.Sedaghati, S.Y.Yang, A.Mosahebi, M.S.Alavijeh, A.M.Seifalian, Biotechnol. Appl. Biochem. 2011, 58, 288. [2] K.Chaloupka, M.Motwani, A.M.Seifalian, Biotechnol. Appl. Biochem. 2011, 58, 363. [3] P.Jungebluth, E.Alici, S.Baiguera, B.K.Le, P.Blomberg, B.Bozoky, C.Crowley, O.Einarsson, K.H.Grinnemo, T.Gudbjartsson, G.S.Le, G.Henriksson, O.Hermanson, J.E.Juto, B.Leidner, T.Lilja, J.Liska, T.Luedde, V.Lundin, G.Moll, B.Nilsson, C.Roderburg, S.Stromblad, T.Sutlu, A.I.Teixeira, E.Watz, A.Seifalian, P.Macchiarini, Lancet 2011, 378, 1997. [4] R.Y.Kannan, H.J.Salacinski, P.E.Butler, A.M.Seifalian, Acc. Chem. Res. 2005, 38, 879.

NANO-165 Monitoring virus viability using dielectrophoretic collections K. Schlegel1,2, D. J. Bakewell3, J. Bailey1, B. Webb4, T. Mukhopadhyay2 and D. Holmes1 1London Centre for Nanotechnology, University College London, UK. 2Department of Biochemical Engineering, University College London, UK. 3Department of Electrical Engineering and Electronics, University of Liverpool, UK. 4Department of Virology, University College London, UK.

Label-free virus detection, quantification and characterisation using electrical methods represents an enabling technology. Applications include the development of powerful point-of-care diagnostics and new process analytical technologies (PAT) for improved bioprocesses manufacture of viral vaccines and gene delivery vectors. Current methods of virus quantification use plaque assays or TCID50 studies, which require 3-14 days per assay. Label based assays are available, such as, fluorescence focus assays, FACS, ELISA or qPCR. These methods require a minimum of 2-24 hours depending on the efficiency of the label. Antibody or PCR based methods do not measure virus particles directly, but approximate virus properties via protein or nucleic acid levels. In this paper we present a novel method to rapidly assay the viability of small volumes of virus containing sample. Dielectrophoresis (DEP) is an alternating current (AC) electrokinetic technique which uses electrodes within a microfluidic flow device to manipulate micro- and nano-scale biological particles suspended in aqueous media [1-3]. DEP collection rates are used to measure the passive electrical properties of virus particles. We demonstrate the utility of using DEP collection rate measurements to discriminate between viable and inactivated bovine herpes simplex virus (BHV-1). [1] R. Pethig, Dielectrophoresis: Status of the theory, technology, and applications. Biomicrofluidics. 4(2): p. 35 (2010). [2] B.H. Lapizco-Encinas and M. Rito-Palomares, Dielectrophoresis for the manipulation of nanobioparticles. Electrophoresis. 28(24): p. 4521-4538 (2007). [3] H. Morgan and N.G. Green, AC electrokinetics. 2003: Research Studies Press, Baldock, England and Institute of Physics Publishing, Philadelphia, USA.

NANO-167 Antisense Gold Nanobeacons for Gene Therapy J. Conde1,2, J. Rosa1,3, M. Larguinho1,3,4, A.R. Fernandes1,5,6, L. Raposo5,6, S. Santos5,6, P.V. Baptista1,*

1CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal. 2Instituto de Nanociencia de Aragón, Universidad de Zaragoza, Zaragoza, Spain. 3REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal. 4BioScope Group, Facultade de Ciencias de Ourense, Universidade de Vigo, Spain. 5Centro de QuímicaEstrutural, Instituto Superior Técnico, Lisboa, Portugal.

6CB3, Faculdade de Engenharia e CiênciasNaturais, UniversidadeLusófona de Humanidades e Tecnologias, Lisboa, Portugal.

Antisense DNA [1]has emerged as a powerful tool for sequence-specific post-transcriptional gene silencing capable ofdownregulating target genesassociated to disease. However, naked DNA oligonucleotides showextremely short half-livesmainly due to nuclease action, poor chemical stability and common dissociation from vector. Gold nanoparticles (AuNPs) showa tremendous potential as intracellular delivery vehicles for antisense oligonucleotides providing for increased protection against nucleases[2] and targeting capability via specific simple surface modification [3]. We constructed an Antisense Gold-nanobeaconconsisting of a stem-looped oligonucleotide double labelled with 3’-Cy3 and 5’-Thiol-C6 complementary to the Kozak consensus translation initiation site and to the start codon of EGFP, and tested for the effective blocking of EGFP expression

in a colorectal cell-line (HCT-116).The beacon’s increase in fluorescence upon hybridisation to the specific targetwas usedto evaluate the effective silencing, as the beacon’ fluorescence increases for increased levels of silencing.An integrated proteomic profile was performed using two-dimensional protein electrophoresis followed by mass spectrometry, which reveals altered expression level of a group of intracellular proteins in presence of the Antisense Gold-nanobeacon. [1] Y. Fichou, C.Ferec. Trends Biotechnol.24, 563-570 (2006). [2] J. Conde, J.M. de la Fuente, P.V. Baptista. Nanotechnology 21, 1-6 (2010). [3]N.L.Rosi, D.A.Giljohann, C.S. Thaxton, A.K. Lytton-Jean, M.S. Han, C.A.Mirkin. Science 312, 1027-1030 (2006). NANO-168 Auricular Reconstructionwith a NanocompositeScaffold L. Nayyer1, C. E. Ramirez1, M. Birchall1, 2, A.M. Seifalian1, 3*, G. Jell1

1Centre for Nanotechnology&RegenerativeMedicine, UCL, Division of Surgery Interventional Science 2ENT Institute, University College London, London, UK 3*Royal Free Hampstead NHS Trust Hospital, London, UK

The use of alloplasticmaterials has greatlyincreased the options available for auricular reconstruction resultingfromcongenitalmicrotia, trauma and cancer. Currenttreatmentinvolves a hand-carvedautologous costal cartilage framework, but drawbacks are associatedwithdonor site morbidity, length of operation and the need for skilled surgeons. Syntheticmaterialscaffoldsoffer an alternative solution, withMedpor® being the currentmarket leader [1]. However, complications associatedwith infection and extrusion canoccur. Herewediscuss the development of a non-biodegradablenanocomposite (NC) scaffoldincorporatingpolyhedraloligomericsilsesquioxane (POSS) nanocagesinto polycarbonate basedurea-urethane (PCU) forauricular reconstruction.Previous studies have shown that auricle-shaped POSS-PCU constructs can be created which, have an elastic modulus similarear native cartilage. Here we report on thehumanfibroblast interactionwiththisconstructincluding, cellularadhesion (staticvs. dynamicenvironments), proliferation, and ECM compared to Medpor®. Significantly higher cell adhesion was found on POSS PCU compared to Medpor® (p < 0,05). Dynamic condition reducedcell attachment on both scaffolds; under this condition POSS PCU maintained higher cell adhesion. There was no significantdifference in cellproliferation or metabolismper cellbetween the materials, but the amount of soluble collagen productionwassignificantly(P< 0.05)increased on POSS-PCU compared to Medpor®. These are promisingresults but furthercellstudies (currentlybeingcompleted) on cell infiltration into the scaffold and angiogenesismay lead to futherscaffold optimisation. [1] T. Wellisz, Plast. Reconstr. Surg., 91, 811-18 (1993)

NANO-169 Electronic properties calculation and docking studies toward new potentially AChE inhibitors N. Kitisripanya1 and S. Hannongbua2

1 Faculty of Science and Engineering, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Thailand. 2 Chemistry Department, Faculty of Science, Kasetsart University, Bangkok, 10900. Thailand.

The main purpose of this study was the use of the couple of natural coumarin, benzopyrone found in many plants and donepezil as lead materials for generating new potentially candidates of acetylcholinesterase

inhibitors. Therefore, we studied the electronic structure of 26 molecules derivatives from the couple using the following groups: methyl, hydroxyl, and acetyl. All structures were fully optimized by HF method at the basis set 6-311G(d,p). The single point electronic properties example for highest occupied molecular orbital (HOMO), HOMO - 1, lowest unoccupied molecular orbital (LUMO), and LUMO +1, were calculated by B3LYP at the basis set 6-311G(d,p). In order to compare the electronic properties of 26 compounds with donepezil, Principal Components Analysis (PCA) were used. Autodock tool also was used for study the binding in the active site gorge. Compound 18 and 19 have the most closest correlation with the compound 20 which have the best estimated binding energy. Furthermore, the docking method was perform to predict the binding in the AChE pocket. From both performed analysis it was possible to devise some structures that are better correlated to donepezil. [1] H.K. Dong, et at, Life Science, 80, 1944 (2007). [2] A.A.N. de Paula, J.B.L. Martins, R. Gargano, M.L. dos Santos, L.A.S. Romeiro, Chem. Phys. Lett., 446, 304 (2007).

NANO-170 Preliminary biological assessment and bacterial adhesion of lanthanides doped hydroxyapatite composites for bone tissue engineering applications Soraia Fernandes1, N. Sooraj Hussain1,2*, P. S. Gomes3, M.A. Lopes4, M. H. Fernandes3 and J.D. Santos4

1INESC Porto, Rua do Campo Alegre, 687, 4169-007 Porto, Portugal. 2Departamento de Física, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, 687, Portugal. 3Laboratório de Farmacologia e Biocompatibilidade Celular. Faculdade de Medicina Dentária, Universidade do Porto, Rua Dr. Manuel Pereira da Silva, 4200-393 Porto, Portugal 4DEMM, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.

Hydroxyapatite (HA) substituted with rare-earth ions (Lanthanides) is in the focus of interest, for bone-related biomaterial applications [1]. Due to their resemblance to calcium, lanthanides present a pronounced biological activity, as they are able to replace Ca2+ in structured molecules [2-4]. Barta et al. have investigated the use of lanthanides for the management of bone resorption disorders, showing that lanthanides can replace calcium in bones and cause local activation of osteoblasts, which are thus responsible for bone formation [5]. Additionally, lanthanides have been shown to be effective within the management of bone-related inflammatory conditions. Due to the ability to modulate the bone regeneration process, the inclusion of RE elements on the composition of biomaterials for bone tissue regeneration has been assayed. Some approaches include the development of bioactive glasses containing lanthanides which, when added to the HA matrix, allow for the development of a bioactive composite material, with improved biomechanical properties and the ability to release, in a controlled way, bioactive species, including lanthanides. Hence, this work reports the preparation and characterization of lanthanide doped HA composites. The structure and morphology of these composites have been investigated by XRD, FTIR, and SEM with an energy dispersive analyzer (EDS). In the preliminary work, bacterial adhesion was studied by MTT assay for the first 90 minutes of contact with the materials, with three different strains: S.Aureus, S.Epidermidis and Pseudomonas Aeruginosa. Biological properties of the developed composites were studied also by an MTT test in order to evaluate the biocompatibility and toxicity. The accomplished results demonstrate that the studied material has potential to decrease bacterial adhesion to the surface and is non-toxic to osteoblast-like cells, and further optimized could be interesting to be used in bone tissue engineering applications. [1] A. Aissa, et al., Sorption of tartrate ions to lanthanum (III)-modified calcium fluor- and hydroxyapatite. Journal of Colloid and Interface Science, 330(1), p. 20, (2009). [2] S.P. Fricker, The therapeutic application of lanthanides. Chem. Society Reviews, 35(6): p. 524, (2006).

[3] K.H.Thompson, K.H. and C. Orvig, Editorial: Lanthanide compounds for therapeutic and diagnostic applications. Chem. Society Reviews, 35(6), p. 499, (2006). [4] T. Matsuda, C. Yamanaka, and M. Ikeya, ESR study of Gd3+ and Mn2+ ions sorbed on hydroxyapatite. Applied Radiation and Isotopes, 62(2), p. 353, (2005). [5] C. Barta, et al., Lanthanide containing compounds for therapeutic care in bone resorption disorders. Dalton Trans, 43, p. 5019, (2007).

NANO-171 Fractal Dimension of Mucoadhesive Polymers/Drugs (Alcohol/Aniline Analogues) F. Torrens1, G. Castellano2

1Inst. Ciència Molecular, Universitat València, P. O. Box 22085, 46071 València, Spain. 2Cátedra Energesis, Universidad Católica de Valencia, 46001 València, Spain.

Mucoadhesive polymers are used in pharmaceutical formulations to elaborate drugs applicable in mucous zones, e.g., gastrointestinal/vaginal tracts, ocular mucous membrane and nasal cavity. They can interact and become fixed to mucus via mechanisms, e.g., molecular interpenetration, van der Waals forces, hydrophobic interactions, electrostatic forces, hydrogen bonds, etc., which allows augmenting the residence period in the organism and increasing the bioavailability of the drugs that they deliver. The physicochemical properties of drug/polymers, e.g., molecular weight, grade of ionization, concentration and polymer swelling kinetics, affect formulation mucoadhesion magnitude, rheological behaviour and drug absorption. Generalized valence charge-transfer indices were applied to the calculation of the molecular dipole moment of homologous series of transdermal-delivery drug models: percutaneous enhancers phenyl alcohols/4-alkylanilines. Drug fractal dimension was examined [1,2]. Here, method is extended to drugs/polymers. Hyaluronan is selected as mucoadhesive/biodegradable polymer. It is chosen aniline analogue minoxidil and alcohol analogues (steroid betamethasone, eugenol, diclofenac) as model drugs. Conclusions follow. (1) Hyaluronan is an important component of articular cartilage, where it is present as a coat around each chondrocyte. When aggrecan monomers bind to hyaluronan in the presence of link protein, large highly anionic aggregates form, which imbibe water and are responsible for cartilage resistance to compression. The molecular weight of hyaluronan in cartilage decays with age but the amount increases. (2) Polymer rheological behaviour formulated in pH 4–7 does not differ, which is proper of unstructured systems. pH < 4 generate gels because of hydrophobic interactions/hydrogen bonds; gels result promising for administration on skin/mucous membranes. [1] F. Torrens, Leb. Sci. J., 5(1), 61 (2004). [2] F. Torrens and G. Castellano, J. Liq. Chromatogr. Relat. Technol., 31, 2337 (2008).

NANO-172 Morphology Analysis of phospholipids liposome deposited onto a solid surface – a tool to understand how to maintain liposome integrity at an interface A. A. Duarte1, J.T. Marquês2, A. S. Viana2, M. Raposo1 1CEFITEC, Departamento de Física, Faculdade de Ciências e Tecnologia, FCT, Universidade Nova de Lisboa, Portugal. 2Centro de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Ed. C8, Campo Grande, Portugal.

Phospholipid liposomes find several applications, including incorporation of pharmaceutical drugs or biomolecules for drug delivery and also modelling cell membranes. Liposomes formed from self-assembly of lipids are important cell membrane models since lipids are basic building blocks of cells. The characterization of formation of layer-by-layer (LbL) films from phospholipid liposomes can give information which can be used in

liposome applications. For example obtaining information on the true conformation of the liposome may be crucial for interpreting data on the films or for specific applications since during adsorption liposome may remain intact in the LbL films [1, 2] or suffer rupture to form bilayers [3]. In this work we used atomic force microscopy at liquid interface to determine the morphology of 1.2-dipalmitoyl-sn-Glycero-3-[Phospho-rac-(1-glycerol)] (Sodium Salt) (DPPG) layers adsorbed onto LbL films with poly(allylamine hydrochloride) (PAH). The obtained topographies were analyzed using functional or statistical parameters and functions which give information about the surface structure allowing infer about the structure of liposome adsorbed. In this study, the power spectral density function, which provide quantitative information about roughness, growth regime, grains size and correlation length, was analyzed having as variable the adsorption time of DPPG liposome onto PAH already adsorbed molecules. The found results of roughness and grain size were compared with DPPG adsorbed amount obtained from adsorption kinetics curves obtained from quartz crystal microbalance and vacuum ultraviolet spectroscopy. So, this work led us to conclude that both electrical charge and surface roughness can control the rupture of liposome. [1] V.P.N. Geraldo, M.L. Moraes, V. Zucolotto, O.N. Oliveira Jr., Journal of Nanoscience and Nanotechnology, 11, 1167 (2011). [2] M.L. Moraes, M.S. Baptista, R. Itri, V. Zucolotto, O.N. Oliveira Jr, Materials Science and Engineering, 28, 467(2008). [3] S. Gromelski, A.M. Saraiva, R. Krastev, G. Brezesinski, Colloids and Surfaces B-Biointerfaces, 74, 477 (2009).

NANO-173 Continuous flow production of cationic liposomes at high lipid content in microfluidic devices for gene delivery and vaccine therapy

T. A. Balbino1, N. T. Aoki1, A.R. Azzoni2, L.G. de La Torre1

1University of Campinas, Campinas, SP, Brazil. 2University of São Paulo, São Paulo, SP, Brazil.

Liposomes are amphiphilic lipid structures, which in aqueous media self-assemble into spherical bilayers with an aqueous interior lumen. The use of cationic lipids allows their electrostatic complexation with DNA and then makes possible efficient gene delivery inside cells. However, production technologies that use non toxic solvent and precisely control the liposome size at high lipid content are still a challenge. Microfluidics is a powerful technology that allows the production of liposomes by the hydrodynamic focusing method [1] and also cationic liposomes. Firstly, we studied a microfluidic device with the single hydrodynamic focusing method. This method uses a central stream with the lipid dispersion in ethanol, which is injected in the middle stream and hydrodynamically compressed by two aqueous streams. The ethanol diffusion from the inner stream to the aqueous stream allows the liposome formation. In order to intensify the mass diffusion, aiming at the increase of the surface area between the two fluids, a second device was designed with double hydrodynamic focusing. We investigated the influence of the parameters Fluid Flow Velocity, Flow Rate Ratio and Total Lipid Concentration on particle size. The biological efficacies of cationic liposomes produced by both microfluidic devices were in vitro examined in HeLa cells, which proved their feasibility for gene delivery and vaccine therapy applications. Thus, the technology employed in the present study is particular useful for cationic liposome production, given the continuous processes advantages and the reduced number of steps. [1] A. Jahn, S. Stavist, J. S. Hong, W.N. Vreeland, D. L. Devoe, M. Gaitan, ACS Nano, 4, 2077 (2010).

NANO-174 Continuous Formation of DNA/Cationic Liposomes Complexes using Microfluidic Hydrodynamic Focusing Devices for Gene Delivery and Vaccine Therapy T. A. Balbino1, A. R. Azzoni2, L. G. de La Torre1

1University of Campinas, Campinas, SP, Brazil. 2University of São Paulo, São Paulo, SP, Brazil.

The spontaneous complexation process between cationic liposomes (CL) and DNA generates aggregates of different characteristics, and it depends of several factors. The complexation inside microfluidic devices is a promising perspective of controlling the size and polydispersity of these complexes, which could reflect in differences for transfection processes. Aiming to evaluate the process parameters of these complexes production in microfluidic systems, we studied two microfluidic devices: one with a simple straight hydrodynamic flow focusing and a second one with barriers in the mixing microchannel (patterned walls). The conventional bulk mixing method was used as comparison. The results showed that the incorporation of the DNA into the liposomal structures was different for both microfluidic devices; the temperature influenced the average size of complexes produced by the simple microfluidic device, while it did not influence in the patterned walls. The differences were also observed in DNA probe accessibility into the complexes. The simple microfluidic device yielded similar quantity of non-electrostatic bound DNA to that of the bulk mixing method. Interestingly, the complexes produced by the patterned microfluidic device presented DNA probe accessibility decreased in 40% and achieved lower in vitro transfection levels in HeLa cells than the bulk mixing and simple microfluidic complexation methods. These differences are probably consequence on different association between DNA and cationic liposomes controlled by the microfluidic devices. This study contributes to the development of rational strategies in controlling the formation of DNA/cationic liposomes complexes for further applications in gene and vaccine therapy. NANO-175 Development of a T2 contrast agent for magnetic resonance imaging using ultra small super paramagnetic iron oxide nanoparticles conjugated with Anti-MUC1 S. Shanehsazzadeh1,2, M. A. Oghabian1,2, F. Johari Daha3, A. Lahooti1, M. Amanlou 4, B.J. Allen5

1Department of Medical Physics and Biomedical Engineering, Faculty of Medicine, Tehran University of Medical Science. 2Research Center for Sciences and Technology in Medicine, Emam Hospital, Keshavarz Blvd, Tehran14197. 3Nuclear Science Faculty, Nuclear Science& Technology Research Institute, Tehran, Iran. 4Department of Medicinal Chemistry, Faculty of Pharmacy and Drug Design and Development Research Center, Tehran University of Medical Science. 5Head, Experimental Radiation Oncology Cancer Pathology & Cell Biology Laboratory Ingham Institute of Applied Medical Research Discipline of Pathology School of Medicine University of Western Sydney NSW Australia

We prefer to make an oral presentation in Diagnosis and Imaging section. MUC1 antigen is a high-molecular-weight glycoprotein that is aberrantly overexpressed in human breast and other carcinomas. C595 a monoclonal antibody (mAb) against the protein core of human urinary epithelial mucin commonly expressed in breast carcinomas. The aim of this study is to conjugate ultrasmall superparamagnetic iron oxide nanoparticles (USPIO) with C595 mAb in order to detect MUC1 expression. We have developed a new conjugated NP (USPIO), using C595 antibody for the imaging and therapy of anti-MUC-1-expressing cancers. Briefly the C595 mAb was conjugated to the aminated dextran-coated iron oxide 84 nm particles, one to two mAbs per nanoparticles. After magnetic purification and sterile filtration, PCS tests, immunoreactivity tests, T1 and T2 measurements, cell

binding efficiency test with MDA-MB-231 and MCF-7 breast cancer cell lines and Prussian blue study were carried out. No significant differences were observed in immunoreactivity results between conjugated and non-conjugated mAb. The hydrodynamic sizes of NPs were 84 and 114 nm pre and post conjugation, respectively. The T1 and T2 measurements shows more than 79% and 29% increments (for 0.02 mg/ml iron concentrations) in T1 and T2 values for USPIO-C595 in comparison with USPIO, respectively. The atomic absorption test shows that the conjugated NPs affinity to attached to MDA-MB-231 and MCF-7 were 9.2 and 7.2 times higher than non- conjugated NPs. The signal decrements after incubation of MDA-MB-231 and MCF-7 with USPIO-C595 were -75% and -70%, respectively. This mAb-guided bioprobes (iron oxide nanoparticles) effectively targeted human breast cancer cell lines. Application of USPIO-C595 for imaging of tumor cells could be remarkably advantageous in the diagnosis of cancer with MUC1 over expressions. NANO-176 Evaluation of functional probes for biochips using a novel cassette method V. Zinkevich1, N. Sapojnikova2, S.Alkhalil, M. Sztyler1, J. Mitchell1, T. Kartvelishvili2, N.Asatiani2, I. Matitashvili2

1University of Portsmouth, Portsmouth, PO1 2DT, UK. 2I.Javakhishvili Tbilisi State University, Andronikashvili Institute of Physics, Tbilisi 0179, Georgia.

Biochips are ideal for applications in medical diagnostics. They have a low manufacturing cost, high specificity, flexible probe design, and avoid the costs associated with purchase of commercial microarrays. However, selection of specific genes with similar hybridisation properties can be problematic. We have developed a quick selective method, based on cassette technology where several different probes can be evaluated in equimolar proportions in one reaction. Probes were evaluated using the cassette method and 3-D low-density biochips [1] designed for TORCH infections (Toxoplasma Gondii, Rubella, Chlamydia trachomatis, Cytomegalovirus, Herpes virus), which are commonly encountered in pregnant women. A series of probes for genes associated with particular pathogeneses were designed de novo, synthesized and cloned in a pGEM T Easy vector. Two fragments, cassette N1 (277 bp) and cassette N2 (263 bp), were amplified using T7 and SP6-Cy3 tagged primers and used to evaluated the efficiency of hybridisation using a Portable Imager 5000 (Aurora Photonics, USA) on a biochip [2]. The N1 cassette, comprising probes for four pathogens, exhibited a similar hybridisation capacity for three probes, whereas the fourth probe failed to detect its antisense strand under the conditions used, suggesting that this probe should be re-designed. In contrast, the N2 cassette contained three different probes for the same pathogen. The hybridisation capacity for this cassette was variable, with the different probes exhibiting different hybridisation profiles. This provided the opportunity to select the probe with the best hybridisation profile for the biochip. The main advantage of this approach to probe design is the ability to distinguish hybridisation capacities for different probe sequences present in one long DNA fragment. [1]. S.S. Bavykin, J.P. Akowsi, V.M. Zakhariev, V.E. Barsky, A.N. Perov and A.D. Mirzabekov, Appl. Envir. Microbiol. 67, 922 (2001). [2]. H.K. Yoon, J.S. Kim, I.H. Chung, S.Y. Lee, J.R. Han, C. Park and S.Y. Hwang, BioChip J., 4, 105 (2010).

NANO-177 Nanoparticle induced magnetic field hyperthermia combined with radiotherapy enhances tumour therapy by modulating antihypoxia and vascular disruption P.S. Jiang1,2, H.Y. Tsai2, P. Drake1, C.S. Chiang2 1Industrial Technology Research Institute, HsinChu, Taiwan. 2National Tsing Hua University, HsinChu, Taiwan

Gd doped iron-oxide nanoparticles (GdIONP) were developed for use in tumour therapy via magnetic field hyperthermia (MFH). The effect of the Gd3+ dopant on the particle size and magnetic properties were previously reported. The specific power adsorption rate (SAR) determined with a field strength of 246 Oe and 52 kHz was 36 Wg-1[Fe] for Gd0.02 Fe2.98O4, about 4 times higher than the reported SAR values for Fe3O4. In this study, the potential for GdIONPs in tumour therapy, particle tracking, and thermal ablation, in vivo were demonstrated. Using an intramuscular TrampC1 tumour model, the GdIONPs that accumulated in the tumour during treatment were tracked by T2-weighted MR imaging. Histochemical staining revealed different thermal effects, thermal ablation with necrosis around the intratumour injection region and mild hyperthermia at locations away from this region. We also evaluated the anti-tumour effects of combined GdIONP mediated hyperthermia with radiotherapy. Tumour re-growth delay was estimated by measuring tumour size after treatment with radiation, hyperthermia, hyperthermia followed by radiation and a control. A significant difference (p <0.05) in the ‘tumour volume doubling time’ was observed among the control group (3 days), hyperthermia group (7 days), radiation only group (9 days) and combination treatment group (14 days). Immunofluorescence staining for hypoxia, vasculature and proliferating cells revealed a reduced hypoxia region with vascular disruption and extensive tumour necrosis via thermoradiotherapy. The results indicated that GdIONP mediated hyperthermia improved the efficacy of radiotherapy by the dual function of anti-hypoxia and localized vascular disruption. NANO-178 Nanoparticles for Tolerogenesis: Analyzing Key Properties A. Tekrony1, U. Al-Atar2, V. Wright2, A. Cooper2, D. Cramb1, J. Buriak2, L. West3

1University of Calgary, Calgary, AB, Canada, T2N 1N4 2National Institute of Nanotechnology, Edmonton, AB, Canada, T6G 2M9 3University of Alberta, Edmonton, AB, Canada, T6G 2E1

It has been shown that up until a certain age, infants do not produce the same immune response to foreign ABO blood group antigens as older individuals who have a fully developed immune system. Therefore, organs from donors of incompatible blood types can be transplanted into infants safely, without rejection. ABO-incompatible infant heart transplantation has been shown to result in development of immune tolerance to donor ABO antigens, which is likely associated with persistence of donor antigens. Therefore, we hypothesized that intentional introduction of synthesized ABO antigens to infants would also induce tolerance. Inducing tolerance to foreign ABO antigens in this manner would extend the window of time during which safe transplantation from ABO-incompatible donors could be performed, thus expanding the potential donor pool for infants and children who have passed the age at which their immune system matures. A proposed method to induce tolerance is by conjugating antigens to synthesized silica nanoparticles for introduction to immature individuals, which will allow maximum exposure of circulating lymphocytes to the antigens. Here, characterization of the newly synthesized nanoparticles is shown. The nanoparticles are first characterized in solution using fluorescence correlation spectroscopy (FCS) to determine brightness and tendency to aggregate. Next, bright, non-aggregating nanoparticles are injected into the chorioallantoic membrane of chicken embryos to determine detection

efficiency, aggregation, and uptake characteristics using FCS. It has been found that the PEG-coated silica nanoparticles with a core-shell synthesis can be easily detected and be made monodisperse with few aggregation tendencies in the 100-200 nm range. Uptake into blood vessel walls has not been observed for these nanoparticles over 100nm, indicating that the particles can circulate for a prolonged period of time in the blood stream, thus enhancing the likelihood of successful tolerance induction. NANO-179 Critical factors in nanoparticle drug delivery design K.L. Yaehne1, A.D. Tekrony1, A. Clancy1, Y. Gregoriou1, J. Walker1, K. Dean1, T. Nguyen1, A. Doiron2, K. Rinker2, X. Y. Jiang3, S. Childs3, D.T. Cramb1

1Department of Chemistry, 2500 University Dr NW, University of Calgary, Calgary, Alberta, Canada, T2N 1N4 2Department of Chemical and Petroleum Engineering, Schulich School of Engineering, 2500 University Dr NW, University of Calgary, Calgary, Alberta, Canada, T2N 1N4 3Department of Biochemistry and Molecular Biology, Faculty of Medicine, Health Sciences Centre, 3330 Hospital Drive, NW, University of Calgary, Calgary, Alberta, Canada, T2N 4N1

Nanoparticles are increasingly used in medical applications such as drug delivery, imaging and biodiagnostics, particularly for cancer. The design of nanoparticles for tumor delivery has been largely empirical, owing to a lack of quantitative data on angiogenic tissue sequestration. Using fluorescence correlation spectroscopy, we have determined the deposition rate constants of nanoparticles into angiogenic blood vessel tissue. We show that deposition is dependent on surface charge. Moreover, the size dependency strongly suggests that nanoparticles are taken up by a passive mechanism that depends largely on geometry. These findings imply that it is possible to tune nanoparticle pharmacokinetics simply by adjusting nanoparticle size. NANO-180 A comparative study on corrosion behavior of pulsed electrodeposited Co and Co/nano-ZrO2 in simulated body fluid (SBF) M. E.Ghajar, S.R. Allahkaram, S.Mahdavi School of Metallurgy and Materials Engineering, University College of Engineering, University of Tehran, Tehran, Iran,

Metallic materials (stainless steels, cobalt and titanium alloys) are generally used as load bearing implants and internal fixation devices. 316L stainless steel is widely used in biomedical applicationsbecause of its lower price as compared with the other biomaterials [1]. However, due to its low corrosion and wear resistance in comparison to cobalt and its alloys, and also better biocompatibility of cobalt alloys [2-4],the aim of this study is the deposition of cobalt and cobalt/ZrO2 nanocomposites on the 316L stainless steel by pulsed electrodeposition method. The surface morphology, cross section, hardness, and corrosion behavior ofCo and Co/nano-ZrO2 films were investigated in simulated body fluid (SBF) using optical microscope (OM), scanning electron microscope (SEM) equipped with energy dispersive X-ray analysis (EDXA), Vickers microhardness tester and polarization techniques, and the results were compared with the substrate (316L).The results indicated that, pulse electroplating parameters influenced the morphology, hardness and thickness of the coatings. A comparison of microhadnessand polarization results revealed that composite coating had the highest hardness values and exhibited enhanced corrosion resistance as compared with the 316L substrate and cobalt films. [1] S.R. Paital and N.B. Dahotre, Mater. Sci. Eng., R, 66, 1 (2009). [2] L. Benea, P. Ponthiaux and F. Wenger, Surf. Coat.Technol., 205, 5379 (2011).

[3] Y. Okazaki and E. Gotoh, Biomater., 26, 11, (2005). [4] B.V. Krishna, W. Xue, S. Bose and A. Bandyopadhyay, ActaBiomater., 4, 697, (2008).

NANO-181 Fabrication and characteristics of Co and Co/nano-TiO2 coatings processed by pulsed electrodeposition S. Mahdavi, S.R. Allahkaram, School of Metallurgy and Materials Engineering, University College of Engineering, University of Tehran, Tehran, Iran,

Stainless steel, Co and Ti based alloys arewidely used as implant materials [1].Among them, cobalt based alloys have good mechanical properties, biocompatibility, and high corrosion and wear resistance [2,3]. Moreover,these properties are enhanced by incorporation of nano-sized inert reinforcing particles [4]. As biological response, corrosion and tribological behavior of biomaterials is mainly determined by their surface characteristics (topography, roughness, chemistry and etc.) [4,5],the aim of this work is the deposition of cobalt and cobalt/nano-TiO2 on the surface of 316L stainless steel by pulsed electrodeposition method. The surfaces of the samples and their cross sections were examined via optical microscopy and by scanning electron microscopy(SEM)equipped with energy dispersive spectroscopy (EDS). Vickers microhardness and tribological behavior of Co and Co/nano-TiO2 films were investigated, and the results were compared with the substrate (316L). The results indicated that, the coatings morphology, hardness and thickness were affected by pulseelectroplating parameters. A comparison of microhardness and wear results revealed that composite coating had the highest hardness values and exhibited enhanced wear resistance as compared with the 316L substrate and cobalt films. The coefficient of friction also changed in different samples. [1] Y. Okazaki and E. Gotoh, Biomater., 26, 11, (2005). [2] S.K. Yen, M.J. Guo and H.Z. Zan, Biomater., 22, 125, (2001). [3] J. Baszkiewicz, D. Krupa, B. Rajchel, J.A. Kozubowski, A. Barcz, J.W. Sobczak and A. Kosinski, Vacuum, 81, 1306, (2007). [4] L. Benea, P. Ponthiaux and F. Wenger, Surf. Coat.Technol., 205, 5379 (2011). [5] S.R. Paital and N.B. Dahotre, Mater. Sci. Eng., R, 66, 1 (2009).

NANO-182 Cells cultured on a biomaterial surface have altered transcriptome and proteome P. Komorowski1,2, M. Walkowiak-Przybyło2, B. Walkowiak2

1 Lodz Regional Park of Science and Technology Ltd., Lodz, Poland. 2 Department of Biophysics, Technical University of Lodz, Poland.

Biomaterials used for medical implants or instruments production can cause numerous undesirable effects in human organism. They may affect cells being in a direct contact with them and can cause changes in genes expression and, as a consequence, also in protein profile of these cells [1]. The aim of the present work was to examine an effect of medical steel 316L, poly-para-xylylene (Parylene) and nanocrystalline diamond (NCD) surfaces on genes and protein expression in human endothelial cell line EA.hy 926. Cells grown on the biomaterial surface were used for isolation of mRNA (transcriptome) and cell proteins and peptides (proteome). The collected mRNA was used for estimation of changes in expression of genes involved in control of cell cycle and synthesis of extracellular matrix proteins. The gene expression experiment was performed with the use of Oligo GEArray microarrays system (SA Biosciences). The collected proteome was separated and characterized with the use of 2D DIGE gel electrophoresis system (GE Healthcare Life Sciences). Selected proteins after excision from the gel were identified by mass spectrometry (ESI Waters Q-ToF Premier Micromass). About 30% of genes

represented on the microarrays showed significantly changed expression induced by contact with artificial surfaces and cell proteome was also significantly altered, mainly by elevation of the number of expressed peptides. Peptides of structural proteins (keratin 10 and 9), cytoplasmic proteins (gamma and beta actin and lactate dehydrogenase), proteins secreted to the outside of the cell (albumin and angiotensinogen) and regulatory proteins (nuclear proliferation antigen) were identified as those of an elevated expression. [1] M. Kaminska, W. Okroj, W. Szymanski, W. Jakubowski, P. Komorowski, A. Nosal, H. Szymanowski, M. Gazicki-Lipman, H. Jerczyńska, Z. Pawłowska, B. Walkowiak. Acta Bioeng Biomech, 11 (3): p. 19-2 (2009).

NANO-183 Synthesis and characterisation of Self-organised TiO2 nanotubes on Ti13Nb13Zr J.M. Hernández-López1, A.Conde1, J. de Damborenea1, M.A. Arenas1

1Centro Nacional de Investigaciones Metalúrgicas, CENIM/CSIC, Madrid, Avda. Gregorio del Amo 8, 28040, Spain.

Pure titanium and Ti6Al4V ELI alloys are widely used as structural and/or functional biomaterials for artificial total hip or knee replacements and dental implants due to their excellent corrosion resistance, biocompatibility, osseointegration and biofunctionality. Initially, standard Ti6Al4V alloy was chosen for medical applications, since there was extensive information available from aerospace applications, particularly on fatigue behavior. However, V-free alloys are being specifically developed for medical purposes due to the toxicity of this element. Different alloying elements such as Fe, Nb, Ta or Mo are used to replace vanadium in the alloy which also

modifies the microstructure. Gradually orthopedic alloys shifted from two phase (+) to metastable type

which present a Young’s modulus closer to that of the bone (10-30 GPa). Particularly promising is the near-

Ti13Nb13Zr which reduces the Young’s modulus to 64-83 GPa from 105-120 GPa for + Ti6Al4V [1]. Ion incorporation into the nanotube formation on titanium alloy has been considered of interest to provide antibacterial properties and to promote osteointegration [2]. This is of special interest to face the infections initiated during the surgery. Preventing the adhesion of bacteria to the implant surfaces is a step forward in pathogenesis of infection, because bacteria that do not adhere are rapidly killed by the immune system [3]. In previous works, the authors demonstrated the ability of anodising process to grow F- doped TiO2 films with bactericide properties on Ti6Al4V [4, 5]. Anodizing of Ti6Al4V in acid electrolyte at controlled voltage proceeds

with formation of nanotubes on -phase and recession of -phase (V enriched) due to partial or complete dissolution [6] leading to a decrease of the mean percentage coverage of surface by laboratory S. aureus and S. epidermidis regarding the non anodised alloy [5]. The fabrication of self-organised TiO2 nanotubes on Ti13Nb13Zr alloy was investigated in this work to study their main features. Morphology, stoichiometry and microstructure of the oxide films grown in an acidic fluorine containing solution were studied by scanning electron microscopy, energy-dispersive X-ray spectrometry and Rutherford backscattering spectrometry. Finally, the electrochemical stability of the oxide was evaluated in vitro by electrochemical techniques. [1] J. Breme, E. Eisenbarth and V. Biehl in Titanium and Titanium Alloys. Fundamentals and Applications. chapter 16, p.423, Christoph Leyens and Manfred Peters, Ed. (WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, 2003) [2] M. Yoshinari, Y. Oda, T. Kato, K. Okuda, Biomaterials, 22, 2043 (2001). [3] M. Katsikogianni, Y.F. Missirlis, Eur Cell Mater, 8, 37 (2004). [4]. M.A.Arenas, A. Conde, J.J. Damborenea, E. Matykina, J.M. Esteban Moreno, E. Gómez Barrena, C. Perez-Jorge Permarch, R. Perez Tanoira. Materiales de Titanio anodizado con Flúor. Patent Application Number: WO 2011/141610 A1, (17-11-2011). [5] C. Pérez-Jorge, Journal of Biomedical Materials Research Part A, 100A(7), 1696 (2012). [6] E. Matykina, J.M. Hernandez-López JM, M.A. Arenas, A. Conde, J. de Damborenea. Electrochim. Acta, 56, 2221 (2011).

NANO-184 Chitosan-capped Gold Nanoparticles as a Biochemical Sensor N.M. Sultan, M.R. Johan Nanomaterials Engineering Research Group, Advanced Research Materials Laboratory, Department of Mechanical Engineering, University of Malaya, Lembah Pantai, 50603 Kuala Lumpur, Malaysia.

Gold nanoparticles are prepared in water via chemical reduction technique by complexation of high molar ratio glutamic acid molecules with gold precursors stabilized by the adsorption of chitosan [1]. Acidic anions (glutamate ions) are expected to cap the nanoparticle surfaces while the polycationic nature of chitosan enables attachment of the polymer to the negatively charged gold nanoparticles surfaces through electrostatic interactions. Preparation of gold nanoparticles capped with chitosan was carried out in a single-pot process. Chitosan provide sufficient steric hindrance for the stability of the colloid and also to functionalize the nanoparticles for use as sensors. In this work, we used chitosan capped gold nanoparticles to detect various concentration of carbon nanotubes (α- CNT), copper oxide (Cu2O) and zinc sulphate (ZnSO4) in solution. The optical absorption spectra were increased and shifted to the longer wavelength once the analyte were added to the chitosan capped gold nanoparticles. The gold sensor was able to detect organic and inorganic elements for different concentration of solution due to the well chelating properties of chitosan and the sensitivity of the optical properties of gold nanoparticles. [1] N, Toshima and T. Yonezawa, New. J. Chem., 22 (11), 1179 (1998). [2] H.C. Warad, S.C. Ghosh, C. Thanachayanont and J. Dutta, in Proceedings of International Conference on Smart Materials, Smart/Intelligent Materials and Nanotechnology, SmartMat-'04, Chiang Mai, Thailand, 203–207, 2004

NANO-185 Tubular Nanocomposite Scaffolds for Applications in Paediatric Tissue Engineering A. de Mel 1,2, A.Chronopoulos1 G.Cittadella1, M.A. Birchall1,2, A.M. Seifalian 1,3

1Centre for Nanotechnology & Regenerative Medicine, Division of Surgery & Interventional Science, University College London, London, UK 2UCL Ear Institute, Royal National Throat, Nose & Ear Hospital, 330 Gray's Inn Road, London, UK 3Royal Free Hampstead NHS Trust Hospital, London, UK

A range of diseases predisposes paediatric patients to damage or loss organs of 3D tubular structures. Autologus tissue transplantation is associated with disadvantages that include donor site morbidity, which is of greater concern in the case of paediatric patients. Therefore tissue engineering of such organs is considered a great advantage. 3D tubular structured organs in common have a supportive smooth muscle cell(SMC) layer and the luminal layer of an epithelium or endothelium. We aimed to establish a proof of concept platform technology for fabrication of tubular constructs ranging from 2-8mm diameter using a POSS incorporated biodegradable polycaprolactone nanocomposite polymer. The tubular structures were tested for their biomechanical parameters, which include tensile strength, suture retention, and burst pressure for a range of fabrication methods employed to have a multilayered structures. The degree of porosities that resulted from the use of porogens was confirmed with conventional porosity measurements, complemented with morphological assessment with scanning electron and confocal microscopy. Mechanically optimal structures were introduced with adipose derived mesenchymal stem cells with TGF-β and BMP-4 for the outermost structure aimed at developing a SMC layer whilst the luminal layer introduced with epithelial cells or peripheral blood derived endothelial progenitor cells. The degree of cell-material interactions was confirmed with SEM and confocal imaging and quantified with a cell viability assay. Biomechanical and cell-material interactions confirmed that

biodegradable POSS nanocomposite can be suitably fabricated to tailor-make tubular structures with multilayered surfaces with suitable mechanical properties, biochemical and topographical features for distinct cell support. NANO-186 Isolation and Typing of Tumour Cells from Images of Aptamer-Functionalized Chips M.A.I. Mahmood1,2,3, U. H. T. Pham2,3,4, Y.-T. Kim3,4 and S. M. Iqbal1,2,3,4,5 1Department of Electrical Engineering, 2Nano-Bio Lab, 3Nanotechnology Research and Education Center, 4Department of Bioengineering, 5Joint Graduate Studies Committee of Bioengineering Program, University of Texas at Arlington, Arlington, Texas 76019, USA

Aptamers are new class of molecules that show similar affinities like antibodies but provide much more flexibility in design and applications. Chip surfaces coated with anti-EGFR aptamers have been used to selectively isolated brain tumour cells with high specificity [1-3]. The tumour cells, when bound to chip surfaces, show distinct morphological patterns and enhanced activity unlike healthy cells. This provides a new cytology tool for early detection of cancer. This paper shows quantification of the cellular features like changes in shape, pseudopod formation, rapid growth, etc. As an example, change in number of pseudopods formed by human glioblastoma cells during incubation has been 2 on average, while healthy cells stay inactive with no such activity. This happens although there may be variations in the overexpression of biomarkers that should have altered the behaviour of the sub-populations of diseased cells. The study shows a significant difference in the interactions of normal and diseased cells on functionalized surfaces. The use of appropriate image processing techniques results in an easy, economical and rapid modality for the detection of cancer cells. It has the potential to serve as an additional tool to support histological findings and to identify tumour cells based on their physical behaviour. The framework can dramatically facilitate intervention and prognosis monitoring of known metastatic cancers. [1] Y. Wan, M.A.I. Mahmood, N. Li, P.B. Allen, Y.-t. Kim, R. Bachoo, A.D. Ellington, and S.M. Iqbal, Nanotextured substrates with immobilized aptamers for cancer cell isolation and cytology, CANCER, 118(4), 1145-1154 (2012). [2] Y. Wan, Y. Liu, P.B. Allen, W. Asghar, M.A.I. Mahmood, J. Tan, H. Duhon, Y.-t. Kim, A.D. Ellington, and S.M. Iqbal, Capture, Isolation and Release of Cancer Cells with Aptamer-functionalized Glass Beads, Lab on a Chip, Accepted (2012). [3] Y. Wan, Y.-t. Kim, N. Li, S.K. Cho, R. Bachoo, A.D. Ellington, and S.M. Iqbal, Surface Immobilized Aptamers for Cancer Cell Isolation and Microscopic Cytology, Cancer Research, 70(22), 11 (2010).

NANO-187 Lomustine Particulate Formulations are Effective Brain Cancer Treatments

F. A. Fisusi1, A. Alzharani2, I. Summers2, J. Moger2, A. G Schätzlein1, I. F UCHEGBU1

1. UCL School of Pharmacy, 29-39, Brunswick Square, London WC1N 1AX, UK 2. School of Physics, University of Exeter, Exeter EX4 4QL, UK

Glioblastoma multiforme (GBM) is the most common and lethal type of brain cancer. GBM is associated with dismal prognosis (5 year survival rates of less than 5%) and treatment is challenging due to the presence of the blood brain barrier. This work is aimed at developing new therapies for the effective treatment of intracranial tumours. A self assembling amphiphilic chitosan based polymer was synthesised and used to prepare a particulate lomustine formulation. Intracranial tumour models were established by orthotopic implantation of human glioblastoma cells (U87 MG cells) in female swiss nude mice and, once tumours were established, mice were administered the lomustine particulate (13 mg kg-1) and lomustine dissolved in an ethanolic- polysorbate 80

vehicle (1.2 mg kg-1). The dose of lomustine administered in the ethanolic formulation was limited by poor aqueous solubility of lomustine. The mean survival time for animals that received the particulate formulation (33.17 days) was significantly (p <0.05) prolonged compared to the mean survival time of untreated controls (21.33 days) and animals that received a low dose ethanolic lomustine formulation (22.5 days). The particulate formulation thus enabled the administration of a higher dose of lomustine which was accompanied by a significant therapeutic benefit. We have provided preclinical proof of concept data for a new brain cancer formulation. Brain cancers are associated with the worst prognosis of any of the cancers. NANO-188 Development of Engineered Nanocarriers for the Treatment of Metastatic Melanoma Victoria Sherwood1, Desire Di Silvio1, Aarthi Devarajan2, Paul McNaughter3, Carl Webster1, Andrew Mayers3, Grant Wheeler2, Francesca Baldelli Bombelli1 School of Pharmacy1, School of Biological Sciences2, School of Chemistry3 University of East Anglia, Norwich, UK

We are developing a targeted thermo-multichemo therapy for the treatment of late-stage cancer, using melanoma as a model system of a highly aggressive metastatic malignancy. Metastatic melanoma is often unresponsive to conventional therapies, exhibiting resistance to a number of chemically distinct compounds. This nanomedicine-based therapy will have a higher therapeutic efficacy compared to conventional anti-melanoma treatments. We have engineered multifunctional, polymer-coated superparamagnetic iron oxide nanoparticles (SPIONs), functionalized with melanoma-specific peptides and loaded with a combination of anti-melanoma drugs. As NPs in a biological environment can be covered by a layer of proteins forming the so-called protein corona complexes, we characterized the structure and composition of these SPIONs in serum [1]. In particular, we investigated the effect of environmental proteins in the interaction between these NPs and lipid bilayers that are used as model of the cell membrane. We are also testing these SPIONs in a multiparametric nanotoxicity assay, developed in our lab and have found them to show no toxicity in either a cell line panel or in developing Xenopus laevis embryos. Overall, the data presented here propose a novel multi-assay methodology to assess the toxicity of engineered nanoparticles for drug delivery with particular focus on the development of multifunctional nanocarriers for the treatment of metastatic melanoma. [1] A. Pitek, D. O’Connell, E. Mahon, M. Monopoli, F. Baldelli Bombelli and K.A. Dawson. PLoS ONE, 7, e40685 (2012).

NANO-189 Design, Simulation, and Test of a Millimeter Power Generator H. Cabra, S. Thomas Department of Electrical Engineering, University of South Florida, Tampa, Florida, USA

The conversion of biomechanical energy to electricity using the clean and free energy produced by a living being is being researched and will be a transformative innovation to energy problems and the invasive and toxic devices used as power supplies. The ability of the human body to produce its own sustainable energy through of a particular bodily process (blood flow, respiratory system, heart beats, diaphragm vibration, artery/vessel pulsation, or muscle movement) can revolutionize the way the health care industry and engineering/science/medical professions approach biomedical innovation. This paper presents the design, simulation, and test of a millimeter axial flux permanent magnet, to be operated as a power generator. These systems have the potential of converting mechanical movement energy, and hydraulic energy (such as flow of

body fluid, contraction of blood vessel, dynamic fluid in nature) into electric energy that may be sufficient for self-powering nano/micro devices and systems. The constructed physical model is approximately 4X larger than the design model, and a small commercial motor assembled to the rotor and casing serves as the permanent magnet generator. This miniaturized generator system can be used as green energy source, and has the potential of transforming or replacing parts of systems that have lifetime limitations (e.g. lithium batteries), size restrictions, and compatibility problems. The model was simulated with a volumetric flow rate of water. The resulting pressure and velocity at this flow rate were simulated and measured, and the result was compared with lab test result. [1] C.-S. Liu, P.-D. Lin, and M.-C. Tsai, “A miniature spindle motor with fluid dynamic bearings for portable storage device applications,” Microsystem Technologies, vol. 15, no. 7, pp. 1001–1007, 2009. [2] C. L. Stanfield and W. J. Germann, Principles of human physiology. Pearson/Benjamin Cummings, 2010.

NANO-190 Antibody-functionalized gold nanoparticles to target tumor angiogenesis S. Lucas(1,*), D. Bonifazi(2), V. Bouchat(1,*), O. Feron(3), L. Karmani(4), R. Marega(2), B. Masereel(4,*), , N. Moreau(1,*), V. Valembois(1,*), T. Vander Borght(5,*), C. Michiels(6,*), B. Gallez(7) (1) Research center in Physics of Matter and Radiation (PMR), Laboratoire d’Analyses par Réactions Nucléaires (LARN), FUNDP University of Namur, Belgium; (2) Laboratoire de chimie organique des matériaux supramoléculaires, FUNDP University of Namur, Belgium; (3) Unité de Pharmacothérapie (FATH), Université catholique de Louvain, Belgium; (4) Département de Pharmacie (DP), FUNDP University of Namur, Belgium; (5) Center for Molecular Imaging and Experimental Radiotherapy (IRME), Université catholique de Louvain, Belgium; (6) Unité de Recherche en Biologie Cellulaire (URBC), FUNDP University of Namur, Belgium; (7) Laboratoire de résonance magnétique biomédicale (CMFA), Université catholique de Louvain, Belgium (*) NARILIS – NAmur Research Institute for LIfe Sciences

In medicine, development of hybrid nanoparticles that can be targeted to vascular, extra-cellular or cell surface receptors is often considered as an attractive solution for cancer detection and treatment. Here, we propose a new method to produce small and biocompatible gold nanoparticles (AuNPs) that specifically target the epidermal growth factor receptor (EGFR), a membrane protein overexpressed in several kinds of solid tumors. Since it is well-known that nanoparticle shape, size and surface coating may influence the biodistribution and pharmacokinetic profiles of nanoclusters, studies were performed with the antibody-functionalized AuNPs in-vitro and in-vivo in murine models xenografted with human cancer cells. The nanosized (2-10 nm) AuNPs are synthesized by plasma vapor deposition (PVD). The surface of these AuNPs is then functionalized by a coating of PPAA (plasma-deposited polyallylamine) before their transfer into solution. The PPAA coating enhances the stability of the AuNPs in an aqueous environment and allows the coupling with antibodies. EGFR was targeted by covalently coupled to Cetuximab antibodies to the amino groups present in the polymeric shell of PPAA-coated AuNPs. Size, morphology, composition and dispersion of free and conjugated PPAA-coated AuNPs in solution were analyzed by Transmission Electron Microscope (TEM), CPS Disc Centrifuge, UV and Atomic Absorption analyses. For the in-vivo studies, the antibody was firstly labeled to 125I or 89Zr before being coupled to the PPAA-coated AuNPs. In-vitro cell surface ELISA assays showed that the Cetuximab linked to PPAA-coated AuNPs preferentially binds to the EGFR overexpressed by A431 cells compared to the non-expressing CHO and EAhy926 cell lines. Moreover, the conjugated PPAA-coated AuNPs is capable to inhibit the EGF-induced phosphorylation of EGFR (at tyrosine 1173) as measured by phosphorylation assays and subsequent Western Blot analysis. In-vivo studies (micro-PET) also showed that tumor uptake was not significantly different between free and nanoconjugated Cetuximab, highlighting the fact that the antibody retains its recognition properties also in-vivo.

All these studies demonstrate the possibility of using the antibody-functionalized AuNPs for human treatments. The Targan project is a Waleo 2 project from the Walloon Region NANO-191 Histopathological Examination of Oral Mucosal Incisions Welded by 980nm Diode Laser -in vivo- B. M. Mirdan Institute of Laser for Postgraduate Studies / University of Baghdad / Iraq 2012.

Continuous wave (CW) 980nm Laser tissue welding has been studied for full thickness incisions on the oral cavity of the rabbit -in vivo-. Surgical incisions were done on the hard palate of the rabbits; the incisions were irradiated with 980nm diode laser, 0.63W as an output power and 27s as an exposure time. The immediate clinical results have shown a hemostasis of the bleeding, while the histopathological examination showed comparable results though it revealed a fastened healing pattern in comparison with the sutured incisions. The immediately laser welded tissues was as a result of the denaturation and coagulation of the tissue proteins via the increase of the tissue temperature due to the laser-tissue photothermal interaction. The use of 980 nm diode laser in soft tissue welding is a successful method of wound closure in oral mucosa with the minimum side effects. NANO-192 Method for Evaluation of Delivery Efficiency of Antineoplasic Drugs C. Luna2, R. Mendoza3, A.S. Vázquez1, J. Silva1, J.A. Gómez1

1Facultad de Ciencias Químicas, 2Facultad de Ciencias Físico-Matemáticas, 3Facultad de Ingeniería Mecánica y Eléctrica, Universidad Autónoma de Nuevo León. Av. Pedro de Alba s/n. San Nicolás de los Garza. N.L. México.

A method to evaluate delivery efficiency of the antineoplasic drug paclitaxel was developed by use of culture cell lines. Human epidermoid carcinoma ATCC cell line A431 and human lymphoblastic leukemia MOLT-4 were employed to perform separate assays. Two different cell populations of each line were cultured in single culture

maintained in Dulbecco’s Modified Eagle Media with 4 mM L-glutamine and supplemented with 10% Foetal Bovi37oC in an incubator with a 95% air and 5% CO2 atmosphere. Leukemia cells were treated equally but cultured in RPMI culture media. Upper chamber was considered has mock exposed and bottom chamber was added with coated magnetite nanoparticles associated to paclitaxel. Magnetite was obtained by Massart modified method to generate nanoparticles of 5 to 20 nm in size. Oleic acid and Pluronic® covered nanoparticles and anchored drug. Cell viability from both chambers was assessed 24 hours after addition of nanoparticles to final concentration of 1 µg/µl in bottom chamber and exposure to a magnetic field of 3800 gauss. Surviving cells were measured by the MTS method and related to the mock cell population by measuring absorbance at 590 nm to establish cell viability with previous microscopic analysis for morphological changes exploration in cells. Cells exposed to magnetic field showed almost null cell death in upper chambers indicating most of the drug delivery in bottom chambers which resulted in higher cell death values. 1 J. Gao, H. Gu and B. Xu. Acc. Chem. Res., 42, 8 (2009).

NANO-193 Fluorescence properties of carbon powder obtained by RF PACVD method R.Woś1, J. Skolimowski2, K. Mitura1,3

1 Technical University of Lodz , Institute of Material Science and Engineering, Lodz,1/15 Stefanowskiego st., Poland 2 University of Lodz, Department of Organic Chemistry, Lodz, 12 Tamka st., Poland. 3Koszalin University of Technology, Institute of Mechatronics, Nanotechnology and Vacuum Technology, Poland.

Positive, documented properties of carbon powder (in vitro, experience the impact of reducing the strength of carbon powder, ascorbic acid [1], and spectrophotometry of MTT viability assessment HUVEC and create ROS and RNS [2]) obtained by RF PACVD were the cause of the attached to the surface antibiotic. To increase the possibility of connecting the first step should be to increase the reaction surface of the powder. One way is to purificate the surface of the reaction by the use of trifluoromethane sulfonic acid. Carbon powder RF PACVD after purification as well as the magic acid after attaching ampicillini treated with selected strains of bacteria (Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739 Candida albicans ATCC 10231.) In interaction with the bacteria studied carbon powder in the light of the UV lamp showed fluorescence properties. [1] K. Mitura, M. Szmidt, J. Sikorska, E. Sawosz, A. Kruk, M. Grodzik, T. Niemiec, Carbon Synthetized by Chemical Vapor Deposition Method Enhances the Activity of Antioxidants, Abstract Book, 28 June - 2 July, 2010, Zakopane, Poland [2] K. Solarska, A. Gajewska, W. Kaczorowski, G. Bartosz, K. Mitura, Effect of nanodiamond powders on the viability and production of reactive oxygen and nitrogen species by human endothelial cells, Diamond & Related Materials 21 (2012) 107–113

NANO-194 Mechanical and Swelling Behaviour of New Hybrid Ceramo-Polymeric bio-nanocomposites R. Aversa, A. Apicella Second University of Naples, Aversa, Abbazia di San Lorenzo 81031, ITALY.

Hybrid ceramo-polymeric nanocomposites based on Hydroxyl-Ethyl-Methacrylate polymer (pHEMA) filled with nanosilica particles are presented as biomimetic-scaffolding materials. Cytotoxicity and Osteoblast cells adhesion tests have shown good material biocompatibility and osteoconductivity [1]. Dynamic Mechanical Analysis (DMA) confirmed the hybrid mechanical behaviour of these nanocomposites. Moreover, this class of material swells in presence of aqueous physiological solution according to limiting Case II sorption mode turning from glassy and rigid to soft and rubbery while presenting a mechanical behaviour, at 5 to 10 % nanosilica volume loadings, that is comparable with that of bone (when glassy) and to that of the cartilage and Ligaments (when rubbery). The mechanical bio-mimicking properties of these hybrid nanocomposite could be used to design bioactive implants. In particular, two biomechanical functions based on material physical and mechanical properties could be attained by the new bio-hybrid nanocomposite [2-3]: - Early fixation by nanocomposite swelling time (6 hrs) and - Bone growth mechanical stimulation. The use of mechanically compatible hybrid hydrogels as scaffolding materials are expected to increase prosthesis adaptation mechanisms introducing active interfaces that improve implant biomimetics while reproducing cartilage and ligaments biomechanical functions. [1] C Schiraldi, A D'Agostino, A Oliva, … R Aversa, M De Rosa, Biomaterials, 25 (17), 3645-3653 (2004). [2] R Aversa, D Apicella, L Perillo, R Sorrentino, F Zarone, M Ferrari, A Apicella, Dental Materials, 25(5), 678-690 (2009). [3] D Apicella, R Aversa, F Ferro, D Ianniello, A Apicella, J. of Biomedical Material Research: Part B, Applied Biomaterials, vol-93(1), 150-163 (2010).

NANO-195 Branched Gold Nanoparticles for Thermal Treatments of Cells. Laura Sironi1, Stefano Freddi1, Michele Caccia1, Paolo Pozzi1, Leone Rossetti1, Piersandro Pallavicini2, Alice Donà2, Elisa Cabrini2, Gualtieri Maurizio3, Ilaria Rivolta4, Alice Panariti4, Laura D'Alfonso1, Maddalena Collini1, Chirico Giuseppe1*. 1. PhysicsDept., Università di Milano Bicocca, Piazza della Scienza 3, I-20126, Milano, Italy. 2. General ChemistryDept., Università di Pavia, Pavia, Italy. 3.Ambiental Sc.Dept., Università di Milano Bicocca, Piazza Scienza 2, I-20126, Milano, Italy. 4. Experimental Med. Dept., Università di Milano Bicocca, via Cadore 8, 20052, Monza, Italy

The heat released by shape anisotropic gold nanoparticles under near infrared irradiation have been proposed for photo-thermal therapies and thermo-controlled drug delivery. In all these applications particular care must be given to control the nanoparticle – cell interaction and the thermal efficiency of the nanoparticles, while minimizing their intrinsic cytotoxicity. We present here the characterization of the cell interaction of newly developed branched gold nanostars, obtained by laurylsulfobetaine-driven seed-growth synthesis. The study provides information [1] on the size distribution, the shape anisotropy, the cellular uptake and cytotoxicity of the gold nanostars as well as their intracellular dynamic behavior by means of two-photon luminescence imaging, fluorescence correlation spectroscopy and particle tracking. The results show that the gold nanostars are internalized as well as the widely used gold nanorods and are less toxic under prolonged treatments. Their two-photon luminescence under irradiation at 800-950 nm indicate that they can be also employed for cell imaging. [1] Sironi, Freddi, Caccia, Pozzi et al. J.Phys. Chem. C, doi: 10.1021/jp305021k, 2012.

NANO-196 Improvements relating to Sunscreen agent comprising a plurality of nanoparticles .R.Saffie-Siebert1, J.W.J.Hamilton2,N.Torabi-Pour1, M. Ahmed1, S. Nabahi1, C. Mahony2.A.Byrne2 and J.A.D.McLaughlin2

1SiSaf LTD, Northern Ireland Science Park, Belfast, BT3 9DT, UK. 2 NIBEC, University of Ulster, Newtownabbey, BT37 0QB, UK.

It has been known for some time that ultraviolet (UV) radiation emitted from the sun can be beneficial but also harmful to humans and animals. Exposure to UV radiation from the sun enables the human body to produce vitamin D, essential for bone strength and prevention of osteoporosis, and deficiency of which has been linked to many cancers, heart attacks, strokes, and adverse immunological effects. Exposure to UV radiation from the sun, however, can also result in harmful effects. Specifically, the sun's radiation comprises mainly UV radiation of wavelength from 320nm to 400nm known as UV-A radiation, with a small component of UVB (280-320 nm) which is up to 1000 times more potent for skin damage. Inorganic pigments such as titanium oxide, zinc oxide and iron oxide are known UV shielding agents. Particles of these compounds have been used to improve protection against UV-A radiation, since they scatter and/or absorb light efficiently in the UV domain. However, the use of these inorganic materials in suncare formulas poses a challenge since they can appear white on the skin, due to the reflection of visible light, which is generally aesthetically unacceptable. Here we provide information on a sunscreen agent, not based on titanium or zinc oxides. This sunscreen agent consists of an inorganic material containing a plurality of substantially regularly-spaced nanopores having at least one nanoscale dimension. The plurality of substantially regularly-spaced nanopores provides an interference filter for UV radiation. The interference of UV radiation with the nano porous sunscreen agent decreases the UV radiation

that passes through the sunscreen. Additionally the porous nature of the nanoparticles allows the addition of bioactive molecules such as Vitamin D, with slow release, to simulate the positive effects of sunbathing whilst reducing the risks of UV exposure. NANO-197 Ti-DLC Coatings on Biomedical Applications H. Y. Ueng1, H. C. Tsou2, W. T. Whang2

1Department of Electronic Engineering, Chang Gung University Tao-Yuan, 333 Taiwan 2 National Chiao-Tung University, Hsin-Chu, 300 Taiwan

Up to now, the biocompatibility of DLC coatings has been proved and reported [1]. However, the practical applications of DLC film were limited due to the poor adhesion between film and substrate. Ti-DLC film was proved strongly adhesion with Ti-related alloy and steel [2]. Such as NiTi alloy coated with DLC film was used in biomedical field due to its unique shape memory effect, super-elasticity and biocompatibility [3]. Besides Ti-DLC film also shows with very low friction, resist wear, exceptionally hardness and bio-inert. Hence, Ti-DLC becomes a promising material on biomedical applications. In this study, Ti-DLC thin films were deposited innovatively by a combined electron cyclotron resonance microwave plasma discharge of C2H2 and metallic magnetron sputtering [1-3]. The effects of negative bias voltage and hydrocarbon flow rate for the properties of Ti-DLC films were investigated. The microstructure properties of DLC film was examined mainly by Raman spectroscopy. The interpretation of spectra features in terms of the position of G-peak, the FWHM of G-peak, and the integrated intensity ratio ID/IG were discussed, which showed highly to correlate with the process parameters as well as the

film’s characteristics used for biomedical applications. For the doping of DLC thin film, the sp2/sp

3 content and

metal/C composition ratio of deposited films were also characterized by XPS and EDX, respectively. In particular,

the hardness of films was strongly relevant to the negative bias voltage and the Sp2/sp

3 content of the DLC films.

In vitro test of DLC layers was arranged using normal human fibroblasts. The study was stressed on cytotoxicity, adhesion and cell proliferation. Correlations between the content of sp3 and sp2 bonds and surface and biological properties of DLC layers are discussed. [1] Wen J. Maa et al., Biomaterials, 28, 1620 (2007). [2] H.Y. Ueng, C.T. Guo, J. Vac. Sci. Technol., B 21, 2540 (2003). [3] J.H. Sui, W. Cai, Diamond & Related Materials, 15, 1720 (2006).

NANO-198 The CSDA based estimation of the nanoscale dose distribution in irradiated medium with embedded nanoparticle G. Chernov1, R. Carrillo-Torres1, M. Barboza-Flores2, R. Melendrez2, M. Pedroza-Montero2, V. Chernov2

1Departamento de Física de la Universidad de Sonora, 2Departamento de Investigación en Física de la Universidad de Sonora, A.P. 5-088, Hermosillo, Sonora, 83190 México.

The use of gold nanoparticles (NPs) in radiotherapy may provide a high dose to the tumor while delivering a reduced dose to surrounding normal tissue. Numerous theoretical studies have used Monte Carlo (MC) simulations to calculate macroscopic dose enhancement through the introduction of NPs. Several works were devoted to simulation of the dose distribution in the vicinity of individual NPs. Despite the fact that MC methods provide precise numerical results, it is desirable to have an analytical solution that, while unavoidably simplifying

the physical model, would provide valuable insight into the nature of the studied problem. In this work we propose an analytical approach, alternative to MC simulations, describing the dose distribution around a NP embedded in a medium. The approach describes the following sequence of events: An incident photon interacts with the NP and creates a secondary electron that travels toward the NP surface. During travel, the electron losses energy and slows down. If the electron energy is sufficiently high, electron crosses the NP surface, continues to travel in the medium and eventually stops. The radial dose distribution around the NP is defined as the average energy deposited in a spherical shell at a certain distance from the NP center normalized to its mass. The continuous slowing down approximation (CSDA) that ignores energy-loss fluctuations is used. It is also assumed that emitted electrons travel in a straight-line path. The approach was applied for the estimation of the dose distribution around spherical gold NPs of different size in water irradiated with monoenergetic X-rays with energies between 3 and 300 keV. NANO-199 Fabrication of Plasmonic Nanopore Formation for Ultrafast Genome Sequencing Seong Soo Choi1, T. Yamaguchi1, Myoung Jin Park1, Nam Kyou Park2,

1Research center for Nano-bio Science, SunMoon University, Ahsan, Korea 2School of Electrical Engineering and GRL, Seoul National University, Korea

In this report, the plasmonic nanopores of less than 5 nm diameter were fabricated on the apex of the pyramidal cavity array. The metallic pyramidal pit cavity can also utilized as the plasmonic bioreactor, and the fabricated Au or Al metallic nanopore can provide the controllable translocation speed down using the plasmonic optical force. Initially, the SiO2 nanopore on the pyramidal pit cavity were fabricated using conventional microfabrication techniques. Then, the metallic thin film was sputter-deposited, followed by surface modification of the nanometer thick membrane using FESEM, TEM and EPMA. The huge electron intensity of FESEM with ~microsecond scan speed can provide the rapid solid phase surface transformation. However, the moderate electron beam intensity from the normal TEM without high speed scanning can only provide the liquid phase surface modification. The EPMA electron beam exposure without high speed scanning presents the reduction of the nanosize aperture down to 10 nm. During these processes, the widening or the shrinking of the nanometer pore was observed depending upon the electron beam intensity. Finally, using 200 keV TEM, the diameter of the nanopore was successively down from 10 nm down to 1.5 nm. The observed optical intensity through the Al nanometer size pore with 103 fold increase (~ microwatt) can provide the plasmonic optical force with nano-Newton. The transmitted optical force from the plasmonic nanopore can be found to be as large as ~ 1 nano-Newton. Considering the ~200 pico-Newton required for the optical trapping force of the micronsize molecule, therefore, the fabricated plasmonic nano-aperture can be utilized as optically driven nanopore device.

NANO-200 Laser Tweezers Induced Heating Effect on Human Living Lymphocytes K. Santacruz-Gomez1,2, Chooi Jing Yuan3, M.C. Iglesias de la Cruz4, M. Pedroza-Montero5, J. García Solé6 and D. Jaque6

1Centro de Investigación en Materiales Avanzados S.C. Ave. Miguel de Cervantes 120. Chihuahua, México C.P. 31109. 2Departamento de Física. Universidad de Sonora, Blvd. Luis Encinas y Rosales s/n. C.P. 83000. Hermosillo, Sonora, México. 3Department of Biological Sciences, Faculty of Science, National University of Singapore. Singapore 117543. 4Departamento de Fisiología, Facultad de Medicina, Universidad Autónoma de Madrid. s/n 29029 Madrid, Spain. 5Departamento de Investigación en Física. Universidad de Sonora, Blvd. Luis Encinas y Rosales s/n. Sonora, México. 6Fluorescence Imaging Group, Departamento de Física de Materiales. Universidad Autónoma de Madrid, Madrid, Spain.

Optical tweezers constitute a very well known tool for analyzing and manipulating individual cells. Such cells can be immobilized in optical traps created by highly focused laser beams. These trapping lasers usually emit in the infrared region so that the cell damage is substantially reduced. In addition the trapping laser produces photo-thermally induced temperature gradients in the optical trap that must be controlled in order to avoid excessive cell heating. In this work we have systematically investigated the photo-thermally induced temperature gradient in and around lymphocyte cells optically trapped by a laser beam at 980 nm. The temperature was obtained from the emission shift of CdTe quantum dots (emission peak at 660 nm and excited by 488 nm) that were added to the lymphocyte bath. The effects of these gradients on the cell morphology have been investigated. Moreover, as the lymphocytes have nearly spherical shape, the trapping forces can be easily determined and so investigating the effects due to temperature dependence of viscosity. NANO-201 Minimally Invasive Biosensors for Health Monitoring S. Anastasova, A. M. Spehar-Deleze, P. Vadgama Queen Mary, University of London, Mile End Road, E14NS, London, UK

A crucial need to understanding the mechanisms that underlie athletic performance is the ability to measure biochemical changes during training, competition and subsequent adaptation, necessitating innovative leaps in biosensor design. Our aim is to develop implantable sensors for continuous monitoring of tissue lactate, glucose, oxygen. The sensor is composed of two parts: a working electrode and a stainless steel. This approach minimises the size and enables operation in two-electrode mode, which is important for implantable devices. As for example lactate is one of the most important biomarkers to evaluate training performance. Measuring blood lactate will give us the degree of anaerobic respiration and its possible underlying physiological mechanisms. As an example, lactate levels have provided an index of aerobic versus anaerobic metabolism linked to muscle performance and with this an individual response to specific training; from target ratio of anaerobic to aerobic metabolism, a coach can determine optimal training parameters. Lactate sensors have wide linear range in buffer extending well outside normal physiological range (0-8 mM). This is achieved using sandwich membrane construction, which protects enzyme and enhances selectivity. We demonstrate the progress made with the development of the oxygen sensor which requires no enzymatic layers, glucose and lactate sensor which require enzyme layer. Preliminary in vivo test were done successfully. The possibility of tuning the biochemical properties of new micro-scale sensor materials will give push for further study of outer protecting polymers that will retain surface areas for maintained diffusive transport to the sensing elements and reduce blocked contact with protein rich tissue matrix environment. This will be the first time that biofouling will be studied at such isolated, low dimensional tips.

NANO-202 C225-Conjugated Polymeric Micelle for Targetable Photodynamic Therapy C.Y. Hsu1, C.L. Pai2, Y.C. Chen1, H.L. Su2, P.S. Lai1,2

1Department of Chemistry, National Chung Hsing University, Taichung 402, Taiwan 2 Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung 402, Taiwan

Photodynamic therapy (PDT) is a light-triggered chemical reaction that generates localised tissue damage for the treatment of malignant or nonmalignant conditions. In the clinic, patients treated with photosensitizers should be kept away from direct sunlight or strong indoor lighting to avoid skin phototoxicity. Increasing the accumulation of photosensitizers in tumor tissue may not only reduce the skin photosensitivity but also increase the therapeutic efficacy of PDT. In this study, a monoclonal antibody cetuximab (C225) that has been clinically utilised as a therapeutic agent for targeted cancer therapy, was conjugated to the surface of poly(ethylene glycol)-b-poly(lactide) (PEG-b-PLA) micelle via thiol-maleimide coupling for tumor targetable chlorin e6 (Ce6) delivery and the anti-tumor effects of the formulated Ce6 were investigated in vitro and in vivo. Our results demonstrate that C225-conjugated Ce6-loaded polymeric micelles (C225-PM/Ce6) with particle sizes around 91 nm were obviously uptaken by epidermal growth factor receptor (EGFR) overexpressed A431 cells comparing with the negative control HT-29 cells analysed using confocal microscope and UV-VIS spectrophotometer. In addition, C225-PM/Ce6-mediated PDT revealed lower IC50 (~0.2 μg/ml) than that of PM/Ce6- or Ce6-mediated PDT in A431 cells. Foe in vivo study, single dose C225-PM/Ce6 had higher anti-tumor effects (tumor growth inhibition % = 84.8) through inhibition of cell proliferation than free Ce6 or PM/Ce6 at an equivalent Ce6 concentration of 2 mg/kg after irradiation (100 J/cm2). No significant adverse effects on the body weight of mice using C225-PM/Ce6, PM/Ce6 or Ce6-mediated PDT were observed in vivo. Thus, C225-conjugated Ce6-loaded micelle with improved selectivity via C225/ EGF receptor interactions has the potential to reduce not only the skin photosensitivity but also the requirement of drug dose for clinical PDT.

NANO-203 Nanofiberous layer containing diamonds for tissue engineering applications J. Soukupová1, J.Horáková1, K. Mitura2,3

1Technical University of Liberec, Department of Nonwovens, Studentská 2, Liberec 46117, Czech Republic 2Koszalin University of Technology, Institute of Mechatronics, Nanotechnology and Vacuum Technology, Koszalin, Poland 3Institute of Materials Science and Engineering Technical University of Lodz, Stefanowskiego1/15, 90-924 Lodz, Poland

The study is focused on fabrication and characterization of nanofibrous scaffold with diamonds produced by detonation method. Nanodiamonds are incorporated into the nanofibers by electrospinning technique that uses high voltage for self-organization of polymer solution and subsequent production of nanofibers [1]. For nanofibrous scaffold with diamonds was used needleless electrospinning setup. The layer was characterised by SEM, FTIR and in vitro tests. [1] D. Lukáš, Physical principles of electrospinning (Electrospinning as a nano-scale technology of the twenty-first century), Vol. 41, Iss. 2, Textile Progress, 2009

NANO-204 Continuous (plastic) flow synthesis (CPFS) of phase pure HA and other calcium phosphate phases (β-TCP, CDHA and biphasic HA/β-TCP) at (near) ambient conditions A.Anwar , J.A. Darr

Clean Materials Technology Group, Department of Chemistry, University College London, London WC1H 0AJ

Synthetic Hydroxyapatite (HA) nanoparticles (<100nm) were rapidly synthesised using a novel two pump continuous (plastic) flow synthesis (CPFS) at 70°C in 5 minutes (residence time) at the conditions of pH 10-11 from aqueous solution of calcium nitrate tetrahydrate and diammonium hydrogen phosphate. The product was obtained as a phase pure material with a stoichiometric Ca:P molar ratio of 1.67, without the need for an ageing step. The lattice constants of HA were similar to reference JCPDS Pattern no. 09-432. No decomposition of HA into other phases was observed even after heating at 1000°C in air for 1h. This observation showed the superior high-temperature stability of such `biomimetic’ HA powders. Other calcium phosphate phases (β-TCP, CDHA and biphasic HA/β-TCP) were also obtained by changing the Ca/P ratio and pH of the precursor solutions. The influence of initial Ca/P molar ratio, pH and precipitation temperature on the phase evolution and crystallinity of the nanopowder were systematically investigated and optimised. The samples were characterized by techniques such as transmission electron microscopy (TEM), BET surface area analysis, X-ray powder diffraction analysis, and FTIR and Raman spectroscopy. The morphology and crystallinity of the precipitates and impurities present fit the requirement for its biomedical applications. Hence the proposed synthesis strategy provides a facile and economical pathway to obtain nano sized HA and other calcium phosphate bioceramics with high purity, suitable size and ultra low level of impurities.