Invasive versus Non Invasive Methods Applied to Mummy Research: Will This Controversy Ever Be...

Review ArticleInvasive versus Non Invasive Methods Applied to MummyResearch: Will This Controversy Ever Be Solved?

Despina Moissidou,1 Jasmine Day,2 Dong Hoon Shin,3 and Raffaella Bianucci4,5,6

!Department of Histology and Embryology, Medical School, National Kapodistrian University of Athens,"#M. Asias Street, !!#$" Athens, Greece$%e Ancient Egypt Society of Western Australia Inc., P.O. Box !&', Ballajura, WA (&((, Australia'Division of Paleopathology, Institute of Forensic Science, Seoul National University College of Medicine,Seoul !!&-")), Republic of Korea*Department of Public Health and Paediatric Sciences, Legal Medicine Section, University of Turin,Corso Galileo Galilei $$, !&!$( Turin, Italy#Center for Ecological and Evolutionary Synthesis (CEES), Department of Biosciences, University of Oslo,P.O. Box !&((, Blindern, &'!( Oslo, Norway(Anthropologie Bioculturelle, Droit, Ethique et Sante, Faculte de Medecine-Nord, Aix-Marseille Universite,!# boulevard Pierre Dramard, !''**Marseille Cedex !#, France

Correspondence should be addressed to Dong Hoon Shin; [email protected] Ra!aella Bianucci; [email protected]

Received "# December $%"&; Accepted $" April $%"'

Academic Editor: Timothy G. Bromage

Copyright © $%"' Despina Moissidou et al. (is is an open access article distributed under the Creative Commons AttributionLicense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properlycited.

Advances in the application of non invasive techniques to mummi)ed remains have shed new light on past diseases. (e virtualinspection of a corpse, which has almost completely replaced classical autopsy, has proven to be important especially when dealingwith valuable museum specimens. In spite of some very rewarding results, there are still many open questions. Non invasivetechniques provide information on hard and so* tissue pathologies and allow information to be gleaned concerningmummi)cationpractices (e.g., ancient Egyptian arti)cial mummi)cation). Nevertheless, there are other )elds of mummy studies in which theresults provided by non invasive techniques are not always self-explanatory. Reliance exclusively upon virtual diagnoses cansometimes lead to inconclusive and misleading interpretations. On the other hand, several types of investigation (e.g., histology,paleomicrobiology, and biochemistry), although minimally invasive, require direct contact with the bodies and, for this reason, areo*en avoided, particularly by museum curators. Here we present an overview of the non invasive and invasive techniques currentlyused in mummy studies and propose an approach that might solve these con+icts.

1. Introduction

Mummies represent a unique source of information aboutpast diseases and their evolution. (e question as to how tobest maintain the integrity of archaeological and anthropo-logical specimens in the course of examining this evidencehas been a major cause for dispute among scholars.

(e advent of non invasive techniques (e.g., X-ray, CATscanning, and MRI) for examining mummi)ed remains

has been a breakthrough in paleopathology as retrospectivediagnoses can now be achieved without dissection.

However, mainly because of the structural di!erencesbetweenmodern and ancient so* tissues, the e,ciency of noninvasive techniques has been questioned repeatedly. Manyscholars insist that an accurate diagnosis can be correctlymade only through direct examination of the corpse (i.e.,autopsy, endoscopy). However, this approach creates concernamong curators and archaeologists.

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Article ID 192829

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Here we address the debate from a broader perspectiveconsidering the advantages and disadvantages of both inva-sive and non invasivemethods and propose the creation of anexamination protocol for the analysis of ancient mummi)edremains based upon strict scienti)c and ethical criteria.

2. The Use of Non InvasiveTechniques in Paleopathology

A new era in mummy studies began when the )rst group ofEgyptian mummies was subjected to computed tomography(CT) in "-.- ["]. Scientists were given the opportunity toinspect the ancient Egyptians’ bodies without resorting tothe use of invasive methods [$, /]. Both hard and so*tissues could be di!erentiated from multiple textile layersand artifacts (amulets, death masks, or portraits) and theirpathologies diagnosed.

Although diagenetic alterations of ancient tissues o*engenerate interpretative biases, CT scans have allowed di!er-entiation between tissue structures and embalmingmaterials.Similarly, antemortem traumas could be distinguished frompostmortem manipulations associated with the embalmingprocess [&], generating greater knowledge about the ways inwhich ancient populations treated and preserved their dead.

Arti)cial mummi)cation is the deliberate act of preserva-tion of a body a*er death ['].(is practice is aimed at slowingand/or halting so* tissues’ degradation [0]. Di!erent types oftreatments (e.g., evisceration, use of natron, and coating withcomplex mixtures with antibacterial and antiputrefactiveproperties) allowed long-term preservation of the Egyptianmummies [., #].

Apart from exceptional cases, in which some steps of themummi)cation procedure were documented (i.e., the co,nof Djedbastiuefankh, Pelizaeus Museum, Hildesheim, LatePeriod; the Rhind Magical Papyrus, ca. $%% BC; three papyriin Cairo, Durham Oriental and Louvre Museums, around "stcenturyAD), the Egyptians did not leavewritten or illustratedrecords of their mummi)cation methods ['].

Gaps in direct evidence have, therefore, been )lledwith information derived from numerous written sources.Herodotus ('th century AD) provided the earliest writtenaccounts of mummi)cation (Book II of %e Histories). (isis coupled with the records of Diodorus Siculus ("st centuryBC) and further augmented by the writings of Porphyry (/rdcentury AD). (ese principal sources have long providedthe basis of modern knowledge about Egyptian mummi)-cation techniques [-]. CT scans have thus helped scientistsand Egyptologists to increase their knowledge, which hadhitherto been biased by the cultural stereotyping of Egypt inclassical sources. New and more detailed knowledge aboutthe evolution of arti)cialmummi)cation has emerged ["%, ""].

Over the last decade, a new generation of CAT scannerswith increased power of resolution has been released andvirtual autopsy has become one of the basic steps in anyscienti)c investigation of mummi)ed remains ["$]. Visual-ization technology is an e,cient tool in hard and so* tissuepaleopathology ["/]. Dental diseases (e.g., severe teeth abra-sion, carious lesions, cists/abscesses, in+ammation, and toothloss) ["&] and many degenerative disorders (e.g., rheumatoid

arthritis of the Iceman ["'], anthropo-paleopathological inEgyptian mummies ["0], bone and so* tissue malignanttumours and/or so* tissue adenomas [".], or atherosclerosis["#, "-]) can now be diagnosed.

(e latest developments inCT resolution (MicroCT) haveeven enabled the observation of architectural structures ofbones [$%].

Variations in wavelength radiation or use of Terahertzimaging have also been applied to mummi)ed remains.Depending on the degree of hydration of a mummi-)ed corpse, this technique enables scientists to distinguishbetween features of so* tissues or bone and various artifacts,identifying objects [$"] wrapped within textiles.

MRI (magnetic resonance imaging) has been similarlyadvantageous, especially in the study of hydrated mummies(i.e., bog bodies, South Korean mummies) [$$]. MRI appli-cation on a ".th century Korean body showed unique clearorgan structures, which could not be visualized by CT [$/].Less satisfactory results were obtained from MRI applied todehydrated and embalmed bodies (i.e., Egyptian mummies)[$&–$0].

Despite the increased use of non invasive techniques,scholars still debate whether virtual inspection should beregarded as the “gold standard” in mummy studies or not.

In general, the CT methodological reference standardsapplied to the study of ancient remains are those determinedfrom living patients [$.]. Any kind of modi)cation to CTscanning methodology (e.g., slice thickness or the introduc-tion of other non invasive methods such as ionizing radiationon mummi)ed cells) is still on an experimental level and isapplied mainly in pilot studies of uncertain potential [$#].

As a result of the di!erences betweenmodern and ancienttissues and the absence of well-established methodologicalstandards in mummy studies, misdiagnosis can occur. Grav-itational force modi)es both the morphology and locationof the organs that elapses between burial and exhumation.Organs are displaced to the dorsal portion and their con-traction is severe. While the di!erence in radiodensity isvery important at a diagnostic level for living patients, radio-density does not di!er from organ to organ in mummies.Diagenetic processes can be misidenti)ed as pathologicalconditions and vice versa [$%]. To avoid misinterpretations,the complementary use of invasive methods (i.e., endoscopy,histology) is of utmost importance, either to support or toreject an initial diagnosis.

Another problem associated with the exclusive use ofnon invasive techniques is the lack of multidisciplinarityin teams involved in the interpretations of the data. Validscienti)c research should include trained radiologists, whoseexperience lies mostly in diagnosing living patients, physicalanthropologists or paleopathologists, and archaeologists whoprovide background information [&, "-, $$, $-].

To some extent concerning the scope of non invasivetechniques applied to the study of ancient mummies gen-erates confusion; therefore, the scienti)c purpose of noninvasive methods o*en loses its meaning.

Museum curators and conservation experts usually preferto resort to CT scanning in order to avoid specimen sampling

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and usually disregard the need for an overall anthropopa-leopathological investigation. As a result, the broader useof non invasive techniques has become a fad, a spectaclemisused by some scientists and curators for “infotainment”or advertising purposes. In many cases, the motivation forthe employment of visual imaging is to take a curious glimpseinside amummy ["$] and perform animated /D rendering forpublic display in exhibits rather than acquire sound scienti)cdata.

3. The Role of Invasive Methods inMummy Studies

Prior to recent advances in paleoradiology, invasive methodswere the only available means of examining anthropologicalmaterials scienti)cally. Precious information about ancientlifestyles and diseases was acquired over decades, allowingscholars to gain a more profound historical and biologicalknowledge about populations of the past. (e )rst invasiveexamination of ancientmummies began during the early "-thcentury, albeit as a form of public entertainment [$]. Manymummy unwrappings were carried out on the basis of merecuriosity and limited scienti)c knowledge. (erefore, manyspecimens were partially or completely destroyed.

Gradually, mummy autopsy became a more meticulouspostmortem and provided scientists with information aboutboth pathologies and possible causes of death [/%].

Pioneering palaeopathologists adapted modern labora-tory techniques to tiny mummi)ed tissue biopsies in orderto identify ancient tissue structures. (ey successfully diag-nosed many diseases (e.g., tuberculosis, atherosclerosis, andparasitic diseases) [/"–/'].

Step by step, scientists have developed new methods tosample inner organ tissues, for example, endoscopy throughnatural ori)ces (i.e.,mouth, nasal cavities, and use of forceps),and have progressively reduced the damage caused to mum-mies [/0, /.].

Where endoscopes could not be introduced throughnatural or postmortem openings, a small perforation wasmade in the mummy’s back [/#], so that tissue samples couldbe taken for histological studies.

As in forensic pathology [/-], microscopic examinationof small tissue biopsies (%.. ! %.. cm) is a requisite to comple-ment non invasive methods because it allows an initial diag-nosis to be precisely con)rmed or in)rmed ["., /', &%, &"].

Similar developments in gas chromatography/mass spec-trometry (GC/MS), isotopic analysis, and synchrotron anal-ysis of minimal amounts of mummy hair have providedremarkable information about the daily lives of ancient pop-ulations within various social classes [&$–&&] and detectedchronic or acute exposure to heavy metals [&'–&.].

Advances in paleoimmunology [&#–'%] and paleomicro-biology through so*/hard tissue analysis and secretion swabsled to the retrospective diagnosis of several pathogens (e.g.,salmonellosis, tuberculosis, malaria, human leishmaniasis,and Chagas disease) in mummies ['"–0.] and revealed someof their evolutionary patterns [0#]. However, not all scientistsagree that it is possible to recover ancient endogenous humanand pathogenic DNAs from Egyptian mummies [0-–."].

Sampling of small skin tissue biopsies (%.. ! %.. cm)and textiles (" ! " cm) proved to be a reliable method forassessing potential biodeterioration of a mummi)ed bodyor its external contamination. Microorganism identi)cationthrough cultivation and molecular techniques is extremelyuseful for conservation purposes and to minimize the risk ofpotential hazards to the public, especially whenmummies areon display [.$].

Biochemical investigations (a combination of gaschromatography-mass spectrometry, GC-MS, and thermaldesorption/pyrolysis, TD/Py-GC-MS) applied to skin andtextiles and to dental calculus provide a plethora of informa-tion concerning the recipes used in embalming procedures[./–..] and the diets of ancient populations [.#, .-].

Nowadays, the use of invasive methods for examiningmummies is widely regarded with skepticism. While someresearchers consider autopsies unavoidable, many considerthem a destructive procedure [#%].

Full autopsy has o*en been performed mainly to seeinside a mummy and take samples for experimental researchrather than obtain con)rmation of a disease tentativelyidenti)ed via a non invasive method.

(e archaeological value of a human/animal specimenmust always be a primary concern, especially when itis on display. When mummies are completely wrapped,fully dressed, and accompanied by funerary equipment, theprospect of a full autopsy threatens their integrity [$%].Whereas CT imaging requires only careful transportationof the mummy, invasive examination is more complex butcan potentially be performed in a manner that respects theintegrity of the corpse [$-].

Equally signi)cant is the ethical issue concerning lack ofrespect for a human body. A mummy is a deceased person,not an artifact, and burial customs should not be ignored[#"]. If this assumption is followed, no sampling or limitedsampling should be allowed in order to respect the deceased,and it is equally true that presenting /D virtual renderingsof undressed dead bodies to a lay public also raises ethicalconcerns.

Questions concerning the analyses of anthropologicalremains have been raised in many countries and these callfor a speci)c set of bioethical guidelines [#$].

(e extent of invasive examination to which ancientmummies should be subjected is the cause of much debate.In the absence of speci)c laboratory guidelines and protocols,there is a lack of consistency; this has allowed people withoutsu,cient if any scienti)c background to decide how valuablesamples should be investigated. In most countries, decisionsrely mostly upon the protocols established by individualinstitutes, museums, or team supervisors. Decisions basedupon such independent judgments may therefore vary fromfull autopsy permission to total prohibition of the use of anyinvasive technique.

4. Discussion and Conclusion

*.!. Is the Examination Method the Real Issue? (e con-troversy over the necessity of invasive versus non invasive

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techniques calls for some appropriate and standardized pro-tocols to be applied to mummy research. (e issue is notthe e!ectiveness of invasive or non invasive studies but theirsuitability for mummy research, which at present does notconsistently achieve scienti)c standards.

Firstly, the purposes of many studies are inconsistent.A mummy must be investigated in order to provide schol-ars with answers related to speci)c biological or historicalquestions. An investigation performed simply to observe amummymacroscopically ormicroscopically is not useful andis not ethical. Curiously, only a limited number of studiesfocusing upon speci)c diseases or historical developmentsin funerary artifact types found upon mummies have beenperformed to date.

Secondly, arti)cial mummi)cation techniques vary con-siderably according to environmental conditions and culturalpractices. Various factors, temperature, humidity, soil acidity,and time, cause various cell system modi)cations; thesecan be pinpointed both through invasive and non invasivetechniques.(e state of preservation, fully intact or partiallypreserved, is signi)cant even for mummies of similar type,which means that every mummy is a unique case.

Despite the numerous studies performed upon mum-mies, methodological consistency and scienti)c comparisonare lacking. Validity of results cannot be cross-checked for thelack of comparative studies and when scientists from variousdisciplines collaborate inmultidisciplinary studies, con+ict ofinterests is not uncommon.

*.$. Mummy Research Guidelines: %e Need for an Inter-national Ethical and Scienti+c Committee. (e aim of thisreview is to show that the current controversy is mainlycaused by a lack of internationally established guidelines inmummy research. (is, in turn, calls for an internationalmummy research protocol to be instituted. Composed ofscholars of high repute, whose integrity is widely recognized,a committee should reestablish a series of priorities in thestudy of mummi)ed bodies.

Firstly, ethical issues should be considered [#/]. Scientistsneed to pay respect to the funerary beliefs of the deceased.With advice from cultural anthropologists, ethnologists, andbioethicists, a speci)c protocol to approach each type ofcultural/religious context should be designed.

Secondly, mummy studies should be allowed for scienti)cand educational purposes but not for business (i.e., publicentertainment or commercialmovies).(e purpose of a givenstudy, either medical or archaeological, should be disclosedbefore any kind of investigation is performed, its value beingwidely recognized by the scienti)c community. Similarly, asmany neophytes approach the )eld without proper training,strict selection criteria should be applied.

Obviously, it is impossible to apply a rigid and in+exiblescienti)c protocol to all mummy cohorts. While some uni-versal principles and rules will apply, technical protocols willnecessarily need to be adjusted depending upon the type ofmummy (i.e., dry or hydrated) and its state of preservation(i.e., fully wrapped, intact, partially destroyed, etc.). Nonethe-less, all parameters used for mummy investigations should beclearly detailed and results fully published.

More transparency should be demanded when geneticstudies are released. Entire datasets should be publishedrather than selected sequences. (is would enable otherresearchers to provide the scienti)c community with theirown interpretations and critical assessments of the data.(eabsence of transparency through selective data publicationonly gives rise to accusations of secrecy that taint the nameof science and reputation of the data.

Along with an international protocol for mummy inves-tigations, the creation of a worldwide network of tissue bankswould be an optimal solution. Scientists could be providedwith samples for laboratory research without frequent exam-ination of the original remains [#&] and their research wouldgenerate a comparative database with which to enable moretargeted scienti)c applications.

Mummies represent the most precious anthropologicalmaterial with which ancient cultures have provided us. Sincemummi)ed bodies attract scientists from di!erent )elds, aninternational protocol is now essential and required urgently.(is protocol should clearly answer three main questions:“Are we showing adequate respect to the corpse we areanalyzing?”, “Which scienti)c hypothesis necessitates ourstudy of mummi)ed remains?”, and “Do we propose to studymummies for scienti)c/cultural purposes or for business?”

With the aim of creating a scienti)c committee and,subsequently, of promoting the standardisation of a bioethicalprotocol onmummi)ed remains, the authors plan to organisea dedicated symposium within the next World Congress onMummy Studies (Lima, July $.–/%, $%"0).

Conflict of Interests

(e authors declare that there is no con+ict of interests.

Authors’ Contribution

Despina Moissidou, Jasmine Day, Dong Hoon Shin, andRa!aella Bianucci all contributed equally to this work.

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