SEM analysis contains researches of tooth enamel surfaces of two populations. First
group of samples is tooth enamel of prehistorically ancestor from Vu~edol and the second
group of samples is enamel of modern Croatian citizen. Even on small number of
human teeth samples from cooperage site of Vu~edol (3,000 BC) and today’s Croatian
people, we can conclude about chewing biometry of prehistorically ancestors and today’s
modern Croatian people, comparing interspecifically the morphology of enamel microdefects.
With the interspecific comparison of morphology changes on tooth occlusal surfaces,
we can connect the size and shape of abrasive particles and diet with microdefects
of tooth enamel

Ana Azinović

Dino Buković

Jadranka Keros

Zoran Azinović

English

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Coll. Antropol. 27 (2003) 1: 381–386UDC 616.314.11-074Original scientific paperSEM Analysis of Tooth EnamelZoran Azinovi}1, Jadranka Keros2, Dino Bukovi}3 and Ana Azinovi}41 Department of Dental Pathology, School of Dental Medicine, University of Zagreb, Zagreb,Croatia2 Department of Dental Anthropology, School of Dental Medicine, University of Zagreb,Zagreb, Croatia3 Department of Dental Prosthodontics, School of Dental Medicine, University of Zagreb,Zagreb, Croatia4 Croatian Restoration Institute, Department of Restoration of Archaeological Founds,Zagreb, CroatiaA B S T R A C TSEM analysis contains researches of tooth enamel surfaces of two populations. Firstgroup of samples is tooth enamel of prehistorically ancestor from Vu~edol and the sec-ond group of samples is enamel of modern Croatian citizen. Even on small number ofhuman teeth samples from cooperage site of Vu~edol (3,000 BC) and today’s Croatianpeople, we can conclude about chewing biometry of prehistorically ancestors and today’smodern Croatian people, comparing interspecifically the morphology of enamel micro-defects. With the interspecific comparison of morphology changes on tooth occlusal sur-faces, we can connect the size and shape of abrasive particles and diet with microdefectsof tooth enamel.Key words: enamel, microwear, dental adaptation, dietIntroductionTaking the fact that there are no twosame histological structured teeth or equal-ly chemically built, we were intrigued bythe comparison of morphological specificsof human enamel during five thousandyears. How much can the results of the re-search and the interspecifical comparisonmake the judgment of some future evolu-tion and involution changes and patholog-ical cases easier? From the time whenman began to grow corn, produce alcohol,cultivate sugar and salt, evolution of diethabits started so fast that our anatomyand physiology could not adapt, so thereare many chronic diseases of modern peo-ple caused by diet disorder: diabetes, can-cer, heart diseases, tooth decay (caries),and periodontal disease1.381Received for publication April 2, 2003Legovi} et al. (1998)2,3 affirmed in theirlongitudinal studies the connection be-tween orthodontics malocclusion with ge-neral body growing and chronically respi-ration disease in Croatian population.Other group of Croatian authors withtheir researches affirmed the role of cer-tain bacteria, Veillonela and Streptococ-cus mutans, in etiology of infective endo-carditis4. The connection between change-able diet habits and the level of dental ad-aptation is the most in a bigger and fasterloss of teeth and abrasive changes on thebiting teeth surfaces. ]atovi} et al. (1998)5affirmed the assumption that abrasionaffects the lower teeth than the upperones. Prpi} Mehi~i} et al. (1988)6 havealso researched the abrasion changes inworkers at the textile industry, and theyaffirmed that the most defects are in theenamel of frontal teeth with clear aes-thetics and functional problems.Dental microdeffects analysis are usedin scientific literature for comparison be-tween the died out mammals and modernpeople in their biometry of chewing. Tea-ford et al. (1996)7 researched the dentalmicrodeffects in three branches of an-thropoids in early Oligocene from Fayum,Egypt. The results affirmed the connec-tion between all the three anthropoid bran-ches and modern primate who mostly eatfruits. The research also discovered veryimportant differences in enamel micro-structure at the three researched bran-ches, and the author concluded that themicrostructure elements with differentdiet are response for difference enamelmicrodefects.Hojo et al. (1989)8 is researching SEMenamel microdefects connected with twopopulations of one archaeological site inwest Japan. SEM analysis shows that thetooth microdefects of late Stone Age hori-zon are much bigger than enamel micro-defects in modern population horizon.Walker’s1 hypothesis from 80’s lastcentury are suggesting that diet habitsall kinds are leaving deep effects on dif-ferent biological aspects. We can see dif-ferent kinds of diet through the amountand shape of microdefects.Harmon et Rose (1988)9 and Pastor(1992)10 are telling us about the role ofdental microdefects in diet reconstruc-tion, the research results are affirmingthe hypothesis of the connection betweenway of diet and enamel microdefects inteeth.Material and MethodsThe aim of this research is to compareultramicroscopic changes of human ena-mel, who lived in Vu~edol in the cupperage (3,000 BC) and today’s Croatian citi-zen. SEM analysis contains research ofenamel surface on the biting surface inthe lower jaw molars of males.In purpose of comparable research weprepared two sample groups of enamel.In the first group there were samples ofocclusal surface enamel of first molar inthe lower jaw from copper age – Vu~edol.The second group samples were preparedof occlusal surface of the first molar in thelower jaw of a modern Croatian citizen.Samples of enamel’s chewing surfaceswere taken for microstructural analysis.A part of samples were treated by 36%ortophosphoric acid during 60 seconds,then washed and dried with compressedair.All the samples were treated in sameway, steamed with layer of gold in a de-vice S 150 Sputter Coater-Edwards. Thesample’s surface is coated with a goldlayer which thickness is 10–15 nano me-ters to get a better electrically conductionof the samples. The samples are re-searched in PLIVA d.d., Department forQuality Control, on a scanning electronicmicroscope (SEM) JSM – 5,800 JEOL, 15kV. SEM analysis are very suitable forsurface morphology research. In this tech-nique the reflected and secondary elec-382Z. Azinovi} et al.: SEM Analysis, Coll. Antropol. 27 (2003) 1: 381–386trons are transformed into electric sig-nals with certain detector. Because theyare electric charged they can be divertedin a focus with an electromagnet and theyare brought over in a cathode pipe wherewe get a picture. To avoid any eventuallyatmosphere effects, those researches aremade in a devoid of air space. The amountof reflected and secondary electrons de-pends on the tension we use, on the detec-tors position and on the samples surface.The picture of sample’s surface based onthe electrons has a deep sharpness andvery clear relief. The results are evalu-ated by computer treated microphoto-graphs, device HEWLETT PACKARD ul-tra VGA 1,600.Results and DiscussionSEM analysis covered the research ofenamel occlusal surface. On the samplesof two groups, results of research aregrouped by the level of abrasive changesof occlusal surfaces. On Figure 1 is micro-graphy of enamel surface in cupper agetooth. (Vu~edol, 3,000 BC). Top of the Fig-ure 1 shows that the enamel surface lookssmoother even on a small enlargement.In the middle there is a line that sepa-rates enamel from dentine surface. Themacroscopic research shows a totally lossof an enamel part on the occlusal surface,so this scanning microphotography ismade on the passing part from enamel tothe exponated dentine on biting surfacesof teeth.Figure 2 shows the same sample butonly the enamel surface on a bigger en-largement. The enamel surface is rough,with surface defects wide fissures andniches, covered with different size andnumber particles, but homogeny shapedwith a base. This figure is suggestinghuge and different types of abrasivechanges on the both biting surfaces. Ourresults correspond completely with onesgiven by M. F. Teaford from 1991 and199411,12.Figure 3 shows enamel surface of to-day’s modern Croatian citizen. Macrosco-pic researches did not discover the ex-ponated dentine of occlusal surface. Theenamel surface is not totally smooth, dif-ferently to Figures 1 and 2, it has lesssurface defects and roughness.The biggest difference is discovered ona big enlargement on Figure 4. Surfacedefects are minimal, fissures are follow-ing the natural shape of enamel prisms,and they cannot be connected with theabrasion happened by changes and de-fects.Figure 5 shows enamel structure ofcupper age tooth. Enlargement is 13,000 and shows many enamel prisms, theirsize and orientation. Also, we may followthe arcade form of enamel prism orienta-tion, with an enamel dark zone on theinterprismatic part. The ratio betweenprisms and the interprismatic space tellsus that enamel contains much more an-organic parts and in the enamel structuremore enamel prisms are represented.Figure 6 shows enamel structure of mod-ern Croatian citizen tooth. Here shouldbe mention that we have prepared sam-ples from Figures 5 and 6 treating themchemically (etching enamel – regular wayof modern laboratory researches of ena-mel and dentine structures). The way theenamel prisms look and its shape afteretching by an acid agents is suggestingthat the enamel prism centers are savedand their shape is not changed, the prism’speriphery which is poor with anorganiccomponents is lost and the interprismaticspace is bigger13.Comparing these two figures of micro-structure changes of enamel prism mor-phology, we can suppose that both sam-ple, despite huge time distance, are verysimilar in their chemistry and structurespecifications of tooth enamel14.383Z. Azinovi} et al.: SEM Analysis, Coll. Antropol. 27 (2003) 1: 381–386There are many different ways to findout our prehistorically ancestor’s diet.Between all methods a significant placetakes:384Z. Azinovi} et al.: SEM Analysis, Coll. Antropol. 27 (2003) 1: 381–386Fig. 3. SEM micrography of enamel surfaceof today’s modern Croatian citizen (110 ).Fig. 4. SEM micrography of enamel surfacefrom Figure 3 (13,000 ).Fig. 5. SEM micrography of etched enamel ofa cupper age human tooth (13,000 ).Fig. 6. SEM micrography of etched enamel oftoday’s modern Croatian citizen (13,000 ).Fig. 1. SEM micrography of tooth occlusalsurface of cupper age human tooth (110 ).Fig. 2. SEM micrography of enamel surfacefrom Figure 1 (13,000 ).1. Interspecific comparison of teeth mor-phology;2. Microwear researches;3. Food remains analysis from archaeo-logical sites;4. Diagnosis of metabolic diseases causedby diet.Interspecific comparison of tooth mor-phology is the base of our study and in-cludes understanding the dental adapta-tion of certain nutrition or diet.If we have in mind that mammals usetheir teeth in many purposes, and thatthe mammal teeth are just a part of di-gestion system, then it’s possible to con-clude about dental adaptation1.ConclusionsSamples used in this study are smallin number, so the results should be takenas preliminary ones. However, we can getcertain conclusions that can be a goodsuggestion for further studies:1. The interspecific comparison of themorphological changes on the surfacesof the teeth between two populationsshows the connection of size, numberand abrasive particles from the foodwith the microdefects of enamel.2. Less evidence of another kind of de-fects (caries, erosion) from Vu~edoltime suggests less cariogene diet byprehistorically ancestors.3. Fast evolution of diet habits of modernpeople was not followed by such fastdental adaptation, so the direct effectis a high incidence of caries of hardtooth tissues and chronically periodon-tal disease.R E F E R E N C E S1. WALKER, A. C., Phil. Trans. R. Soc. Lond., B292,(1981) 57. — 2. LEGOVI], M., L. MADY, S. PELLI-ZZER, Coll. Antropol., 22 Suppl. (1998) 133. — 3. LE-GOVI], M., M. LAPTER, M. [LAJ, J. [TEFANAC-PAPI], Coll. Antropol., 22 Suppl. (1998) 185. — 4.PRPI]-MEHI^I], G., T. MAR[AN, I. MILET], D.BUNTAK-KOBLER, Coll. Antropol., 22 Suppl. (1998)39. — 5. ]ATOVI], A., V. JEROLIMOV, V. BERG-MAN, Coll. Antropol., 22 Suppl. (1998) 193. — 6.PRPI]-MEHI^I], G., D. BUNTAK-KOBLER, S. JU-KI], M. KATUNARI], Coll. Antropol., 22 Suppl.(1998) 241. — 7. TEAFORD, M. F., M. C. MAAS, E. L.SIMONS, Am. J. Phys. Anthropol., 101 (1996) 527. —8. HOJO, T., Scanning Microsc., 3 (1989) 623. — 9.HARMON, A. M., J. C. ROSE, The role of dentalmicrowear analysis in the reconstruction of prehis-toric diet. In: Diet and subsistence: Current archaeo-logical perspectives. (Calgary, 1988). — 10. PASTOR,R. F., J. Hum. Ecol., 2 (Special Issue) (1992) 215. —11. TEAFORD, M. F., Dental microwear: What can ittell us about diet and dental function? In: Advancesin dental anthropology. (New York, 1991). — 12. TEA-FORD, M. F., Evol. Anthropol., 3 (1994) 17. — 13.AZINOVI], Z.: Provjeravanje efikasnosti jetkanjahumane cakline in vitro s obzirom na razli~ite kon-centracije fosforne kiseline. M.Sc. Thesis. (Universityof Zagreb, Zagreb, 1981). — 14. AZINOVI], Z., K.JORGI]-SRDJAK, B. LAZI], Coll. Antropol., 22 Suppl.(1998) 25.Z. Azinovi}Department of Dental Pathology, School of Dental Medicine, University of Zagreb,Gunduli}eva 5, 10000 Zagreb, Croatia385Z. Azinovi} et al.: SEM Analysis, Coll. Antropol. 27 (2003) 1: 381–386SEM ANALIZA ZUBNOG ENAMELAS A @ E T A KSEM analiza obuhva}a istra`ivanja caklinske povr{ine zuba dviju populacija. Prvaskupina uzoraka je caklina zuba prapovijesnog pretka stanovnika Hrvatske, a drugaskupina uzoraka caklina suvremenog stanovnika Hrvatske. Iako na malom broju uzo-raka ljudskih zuba iz bakrenodobnog lokaliteta Vu~edol (3000 g.p.K.) i dana{njegstanovnika Hrvatske, interspecifi~nom usporedbom morfologije caklinskih mikroo{te-}enja mo`emo zaklju~ivati o biometriji `vakanja prapovijesnih predaka i dana{njegstanovnika Hrvatske. Interspecifi~nom usporedbom morfolo{kih promjena okluzalnihpovr{ina zuba mo`e se dovesti u vezu veli~ina i oblik abrazivnih ~estica i prehrane smikroo{te}enjima cakline zuba. SEM analiza je potvrdila da su abrazivne promjene nacaklini zuba ~ovjeka iz Vu~edola opsegom ve}e i ~e{}e nego promjene na caklini da-na{njeg stanovnika Hrvatske.386Z. Azinovi} et al.: SEM Analysis, Coll. Antropol. 27 (2003) 1: 381–386

SEM Analysis of Tooth Enamel

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