Carsten Witzel

Insights from the inside: Histological analysis of abnormal enamel microstructure associated with hypoplastic enamel defects in human teeth

We studied the abnormalities in enamel microstructure associated with enamel hypoplasia in human teeth from the early medieval (5th-7th century AD) cemetery of Barbing, Germany, using light and scanning electron microscopy. The main aim of the study was to test the hypothesis that by analyzing the microstructure of fully formed enamel it is possible to reconstruct the reaction pattern of secretory ameloblasts to stress events leading to enamel hypoplasia. From the histological findings, a sequence of increasing impairment of secretory ameloblast function involving three thresholds was deduced. Surpassing of each of these thresholds is assumed to result in characteristic changes in enamel microstructure attributable to specific functional/morphological alterations of secretory ameloblasts. Based on our results we propose a model identifying the principal factors influencing the reaction of secretory ameloblasts to stress. The present study demonstrates that by including microscopic analysis in the study of enamel hypoplasia, it is possible to obtain a more complete picture of the formation of these developmental defects than is possible by inspection of crown surface features alone, and to draw more substantiated conclusions about the possible nature of developmental defects of enamel.

Reconstructing impairment of secretory ameloblast function in porcine teeth by analysis of morphological alterations in dental enamel.

We studied the relationship between the macroscopic appearance of hypoplastic defects in the dental enamel of wild boar and domestic pigs, and microstructural enamel changes, at both the light and the scanning electron microscopic levels. Deviations from normal enamel microstructure were used to reconstruct the functional and related morphological changes of the secretory ameloblasts caused by the action of stress factors during amelogenesis. The deduced reaction pattern of the secretory ameloblasts can be grouped in a sequence of increasingly severe impairments of cell function. The reactions ranged from a slight enhancement of the periodicity of enamel matrix secretion, over a temporary reduction in the amount of secreted enamel matrix, with reduction of the distal portion of the Tomes’ process, to either a temporary or a definite cessation of matrix formation. The results demonstrate that analysis of structural changes in dental enamel allows a detailed reconstruction of the reaction of secretory ameloblasts to stress events, enabling an assessment of duration and intensity of these events. Analysing the deviations from normal enamel microstructure provides a deeper insight into the cellular changes underlying the formation of hypoplastic enamel defects than can be achieved by mere inspection of tooth surface characteristics alone.

Enamel hypoplasia in molars of sheep and goats, and its relationship to the pattern of tooth crown growth

Enamel is the most highly mineralized and durable tissue of the mammalian body. As enamel does not undergo remodeling or repair, disturbances of enamel formation leave a permanent record in the tissue that can be used for life history reconstruction. This study reports light and scanning electron microscope findings on hypoplastic enamel defects, and on the chronology of crown growth in the molars of sheep and goats. A marked reduction of enamel extension rates in cervical compared with more cuspal crown portions of sheep and goat molars was recorded, with formation of the cervical 25% of the crown taking about the same time as that of the upper 75% of the crown. This explains the more frequent occurrence of enamel hypoplasia in cervical compared with upper and middle crown portions. Regarding the identification of hypoplastic enamel defects by external inspection, our results suggest a dependence on the type of defect and the associated presence of smaller or larger amounts of coronal cementum. Defects considered to reflect a slight to moderate impairment of secretory ameloblast function can normally be correctly diagnosed as they are not occluded by thick layers of cementum. In contrast, defects denoting a severe impairment of enamel matrix secretion can typically not be correctly identified because they are occluded by large amounts of cementum, so that neither depth nor extension of the defects can be assessed on external inspection. In these cases, microscopic analysis of tooth sections is required for a correct diagnosis of the hypoplastic enamel defects.

Lines of Evidence–Incremental Markings in Molar Enamel of Soay Sheep as Revealed by a Fluorochrome Labeling and Backscattered Electron Imaging Study

We studied the structural characteristics and periodicities of regular incremental markings in sheep enamel using fluorochrome injections for vital labeling of forming enamel and backscattered electron imaging in the scanning electron microscope. Microscopic analysis of mandibular first molars revealed the presence of incremental markings with a daily periodicity (laminations) that indicated successive positions of the forming front of interprismatic enamel. In addition to the laminations, incremental markings with a sub-daily periodicity were discernible both in interprismatic enamel and in enamel prisms. Five sub-daily increments were present between two consecutive laminations. Backscattered electron imaging revealed that each sub-daily growth increment consisted of a broader and more highly mineralized band and a narrower and less mineralized band (line). The sub-daily markings in the prisms of sheep enamel morphologically resembled the (daily) prisms cross striations seen in primate enamel. Incremental markings with a supra-daily periodicity were not observed in sheep enamel. Based on the periodicity of the incremental markings, maximum mean daily apposition rates of 17.0 µm in buccal enamel and of 13.4 µm in lingual enamel were recorded. Enamel extension rates were also high, with maximum means of 180 µm/day and 217 µm/day in upper crown areas of buccal and lingual enamel, respectively. Values in more cervical crown portions were markedly lower. Our results are in accordance with previous findings in other ungulate species. Using the incremental markings present in primate enamel as a reference could result in a misinterpretation of the incremental markings in ungulate enamel. Thus, the sub-daily growth increments in the prisms of ungulate enamel might be mistaken as prism cross striations with a daily periodicity, and the laminations misidentified as striae of Retzius with a supra-daily periodicity. This would lead to a considerable overestimation of crown formation times in ungulate teeth.

The ideal subject distance for passport pictures

In an age of global combat against terrorism, the recognition and identification of people on document images is of increasing significance. Experiments and calculations have shown that the camera-to-subject distance - not the focal length of the lens - can have a significant effect on facial proportions. Modern passport pictures should be able to function as a reference image for automatic and manual picture comparisons. This requires a defined subject distance. It is completely unclear which subject distance, in the taking of passport photographs, is ideal for the recognition of the actual person. We show here that the camera-to-subject distance that is perceived as ideal is dependent on the face being photographed, even if the distance of 2m was most frequently preferred. So far the problem of the ideal camera-to-subject distance for faces has only been approached through technical calculations. We have, for the first time, answered this question experimentally with a double-blind experiment. Even if there is apparently no ideal camera-to-subject distance valid for every face, 2m can be proposed as ideal for the taking of passport pictures. The first step would actually be the determination of a camera-to-subject distance for the taking of passport pictures within the standards. From an anthropological point of view it would be interesting to find out which facial features allow the preference of a shorter camera-to-subject distance and which allow the preference of a longer camera-to-subject distance.

Developmental and Post-Eruptive Defects in Molar Enamel of Free-Ranging Eastern Grey Kangaroos (Macropus giganteus) Exposed to High Environmental Levels of Fluoride

Dental fluorosis has recently been diagnosed in wild marsupials inhabiting a high-fluoride area in Victoria, Australia. Information on the histopathology of fluorotic marsupial enamel has thus far not been available. This study analyzed the developmental and post-eruptive defects in fluorotic molar enamel of eastern grey kangaroos (Macropus giganteus) from the same high-fluoride area using light microscopy and backscattered electron imaging in the scanning electron microscope. The fluorotic enamel exhibited a brownish to blackish discolouration due to post-eruptive infiltration of stains from the oral cavity and was less resistant to wear than normally mineralized enamel of kangaroos from low-fluoride areas. Developmental defects of enamel included enamel hypoplasia and a pronounced hypomineralization of the outer (sub-surface) enamel underneath a thin rim of well-mineralized surface enamel. While the hypoplastic defects denote a disturbance of ameloblast function during the secretory stage of amelogenesis, the hypomineralization is attributed to an impairment of enamel maturation. In addition to hypoplastic defects, the fluorotic molars also exhibited numerous post-eruptive enamel defects due to the flaking-off of portions of the outer, hypomineralized enamel layer during mastication. The macroscopic and histopathological lesions in fluorotic enamel of M. giganteus match those previously described for placental mammals. It is therefore concluded that there exist no principal differences in the pathogenic mechanisms of dental fluorosis between marsupial and placental mammals. The regular occurrence of hypomineralized, opaque outer enamel in the teeth of M. giganteus and other macropodids must be considered in the differential diagnosis of dental fluorosis in these species.

Deposition of cellular cementum onto hypoplastic enamel of fluorotic teeth in wild boars (Sus scrofa L.)

The nature of deposits present in hypoplastic defects of fluorotic enamel of wild boar teeth was studied by light microscopy and scanning electron microscopy. The fluorotic enamel showed different developmental abnormalities, denoting a severe disturbance of ameloblast function during the secretory stage of amelogenesis. These abnormalities included the occurrence of grossly accentuated incremental lines with associated zones of aprismatic enamel and the presence of different forms of hypoplastic defects. Two types of deposits were present on the hypoplastic enamel: cellular cementum and posteruptively acquired, presumably partially mineralized dental plaque. Coronal cementum is not normally formed in pig teeth. Presence of this tissue in fluorotic teeth of wild boars is seen as indicative of a premature disintegration of the enamel epithelium prior to the completion of amelogenesis. This was supposed to have resulted in a contact of mesenchymal cells of the dental follicle with the surface of the immature enamel and, in consequence, in a differentiation of these cells into cementoblasts. To our knowledge, this is the first study reporting the formation of coronal cementum as part of the spectrum of pathological changes in fluorotic teeth in a species whose tooth crowns are normally free of cementum.

Cementoblastoma in a red deer (Cervus elaphus) from the Late Pleistocene of Rochedane, France

Only relatively few cases of dental abnormalities in wild ruminants have thus far been described in the paleopathological literature. This study reports a case of cementoblastoma, a benign odontogenic tumor of ectomesenchymal origin, in a red deer (Cervus elaphus) from the Late Pleistocene of Rochedane, a prehistoric site in the French Jura. The tumor was attached to the root of a heavily worn loose left maxillary third molar. CT imaging revealed several radiolucent (former soft tissue) spaces of varying shape and size within the mineralized tumorous mass. Light microscopic analysis and backscattered electron imaging in the SEM showed that the process of dental wear had reached the tumor and that the tooth and the attached tumor had undergone considerable microbial diagenesis. This is the first case of cementoblastoma described in a prehistoric animal and also the first report of this type of odontogenic tumor in a deer.

Mineral Apposition Rates in Coronal Dentine of Mandibular First Molars in Soay Sheep: Results of a Fluorochrome Labeling Study: MINERAL APPOSITION RATES IN CORONAL DENTINE

We studied the spatio‐temporal variation of mineral apposition rate (MAR) in postnatally formed coronal dentine of mandibular first molars from Soay sheep repeatedly injected with different fluorochromes. MAR declined along the cuspal to cervical crown axis, and from early to late formed dentine, that is, from the dentine at the enamel‐dentine‐junction (EDJ) to the dentine adjacent to the dentine‐pulp‐interface (DPI). Highest mean MARs (about 21 µm/day) were recorded in cuspal dentine formed in the period of 28‐42 days after birth. Lowest values (<2 µm/day) were recorded in late‐formed (secondary) dentine close to the DPI. The high MARs recorded in the dentine of the cuspal crown portions enable the formation of a large tooth crown within a relatively short period of less than one year. The established MARs in the dentine of the different crown portions of sheep molars will allow a precise determination of the timing of stress events affecting dentine formation. They are also helpful for devising sampling protocols in studies of trace element or stable isotope distributions in sheep dentine aimed at assessing temporal variation of incorporation into forming dentine. Such data are useful in a variety of contexts, including, for example, the exposure to pollutants and the reconstruction of husbandry practices or feeding regimes. Anat Rec, 301:902–912, 2018.