The Enamel Microstructure of the Early Eocene Pantodont Coryphodonand the Nature of the Zigzag Enamel

An additional method for the investigation of the microstructure of enamel is described using the teeth of Coryphodon, Uintatherium, Entelodon, and Crocuta. Under low magnification natural surfaces or sections of teeth display details of the enamel microstructure when the light guide effect of prisms is used. Under the same low magnification even more details were obtained from sputter-coated surfaces of sections. The method is of particular significance for the investigation of large teeth with thick enamels when structures are somewhat irregular. The new method provides a better general survey, where scanning electron microscope images often show confusing details. The enamel of Coryphodon shows oblique lines of nested chevrons that are similar, to some degree, to the zigzag enamel in Crocuta, but a distinct asymmetry between ascending and descending lineaments was observed. This specific Coryphodon -enamel was also found in Uintatherium and Entelodon. This enamel type, which evolved several times in parallel, cannot be attributed to a specific diet, but must be regarded as one of the several ways to strengthen the enamel against breakage.     

The absence of muscle segment homeobox 2 leads to the pyroptosis of ameloblasts by inducing squamous epithelial hyperplasia in the enamel organ

Muscle segment homeobox 2 (MSX2) has been confirmed to be involved in the regulation of early tooth development. However, the role of MSX2 has not been fully elucidated in enamel development. To research the functions of MSX2 in enamel formation, we used a Msx2^−/− (KO) mouse model with no full Msx2 gene. In the present study, the dental appearance and enamel microstructure were detected by scanning electron microscopy and micro-computed tomography. The results showed that the absence of Msx2 resulted in enamel defects, leading to severe tooth wear in KO mice. To further investigate the mechanism behind the phenotype, we performed detailed histological analyses of the enamel organ in KO mice. We discovered that ameloblasts without Msx2 could secrete a small amount of enamel matrix protein in the early stage. However, the enamel epithelium occurred squamous epithelial hyperplasia and partial keratinization in the enamel organ during subsequent developmental stages. Ameloblasts depolarized and underwent pyroptosis. Overall, during the development of enamel, MSX2 affects the formation of enamel by regulating the function of epithelial cells in the enamel organ.     

On the nature of the opaque and translucent enamel regions of some macropodinae (Macropus giganteus,Wallabia bicolor and Peradorcas concinna)

Summary Teeth of three macropod species, M. giganteus, W. bicolor and P. concinna, have been studied using the techniques of light microscopy, scanning- and transmission-electron microscopy and hardness measurement. Light microscope observations showed that the teeth of these species had a translucent enamel region close to the dentine and an outer opaque enamel region at the tooth's surface. These regions were not related to the presence or absence of tubules which are a characteristic feature of marsupial enamel. Hardness tests showed that the opaque enamel was softer than the translucent enamel. Scanning electron microscope observations revealed that there was no correlation between any particular prism packing or orientation and the opaque and translucent enamel regions. Transmission electron microscope observations showed that the translucent enamel region consisted of well defined prisms and well packed, lath-like crystals, whereas the opaque enamel was disrupted by voids (which ranged in size from enlarged micropores to about 2 m in diameter in extreme cases) between crystals and some randomly oriented, loosely packed crystals. This disruption within the opaque enamel region was more common at prism boundaries but pockets of disrupted enamel were also found within prisms and interprismatic regions. The opacity of the enamel was caused by scattering of light from the voids. The ultrastructure of the opaque enamel region indicated that this region was hypomineralized; hardness tests and polarized light microscope observations were consistent with these results.     

Variation of the Schmelzmuster and Other Enamel Characters in Molars of the Primitive Pliocene Vole Ogmodontomys from Kansas

Variation in enamel microhistology (the schmelzmuster) and other features were examined in adult Ogmodontomys poaphagus molars from the Meade Basin of southwestern Kansas. Measurements of total enamel thickness and thickness of tangential enamel were taken on individual triangles of first upper molar samples from the superposed Ripley B, Hornet and Rexroad 2A local faunas. Although thickness of total enamel layers and the tangential zone were not statistically different between triangles and sites, the quality of tangential enamel on the trailing edges increased, from incipient tangential and well-developed tangential enamel (Ripley B and Hornet) to solely well-developed tangential enamel (Rexroad 2A). The fundamental schmelzmuster of O. poaphagus is radial enamel on the leading edges and radial plus incipient or well-developed tangential enamel on the trailing edges. These results support the assertion that the schmelzmuster is a consistent and genetically controlled character mosaic, and confirm suggestions in the literature that Ogmodontomys is distinct from Mimomys. Paired sample Student’s t-tests of trailing/leading edge samples for each triangle were used to test for differentiation. Although the molars of Ogmodontomys poaphagus appear undifferentiated under the light microscope, and are indeed separable from the distinctly negatively differentiated molars of Mimomys by this technique, our results show that the M1s of O. poaphagus are slightly negatively differentiated, with various triangles being either undifferentiated or negatively differentiated. O. sawrockensis has a more primitive schmelzmuster than O. poaphagus, and the presence of discrete (primitive) lamellar enamel on the apices of at least one triangle in 55% of the sample from Fallen Angel B, combined with the absence of lamellar enamel in molars of O. poaphagus from Fox Canyon and all subsequent Ogmodontomys samples, conclusively demonstrates that the Fallen Angel B population of O. sawrockensis was not ancestral to O. poaphagus in the Meade Basin. A unique mosaic of dental and size features indicates that the early Blancan O. pipecreekensis from Indiana is a sister species to O. sawrockensis and not ancestral to O. poaphagus.     

Fine structural and immunohistochemical detection of collar enamel in the teeth of <Emphasis Type="Italic">Polypterus senegalus</Emphasis>, an actinopterygian fish

This is the first detailed report about the collar enamel of the teeth of Polypterus senegalus. We have examined the fine structure of the collar enamel and enamel organ of Polypterus during amelogenesis by light and transmission electron microscopy. An immunohistochemical analysis with an antibody against bovine amelogenin, an antiserum against porcine amelogenin and region-specific antibodies or antiserum against the C-terminus, middle region and N-terminus of porcine amelogenin has also been performed to examine the collar enamel matrix present in these teeth. Their ameloblasts contain fully developed Golgi apparatus, rough endoplasmic reticulum and secretory granules. During collar enamel formation, an amorphous fine enamel matrix containing no collagen fibrils is found between the dentin and ameloblast layers. In non-demineralized sections, the collar enamel (500 nm to 1 μm thick) is distinguishable from dentin, because of its higher density and differences in the arrangement of its crystals. The fine structural features of collar enamel in Polypterus are similar to those of tooth enamel in Lepisosteus (gars), coelacanths, lungfish and amphibians. The enamel matrix shows intense immunoreactivity to the antibody and antiserum against mammalian amelogenins and to the middle-region- and C-terminal-specific anti-amelogenin antibodies. These findings suggest that the proteins in the enamel of Polypterus contain domains that closely resemble those of bovine and porcine amelogenins. The enamel matrix, which exhibits positive immunoreactivity to mammalian amelogenins, extends to the cap enameloid surface, implying that amelogenin-like proteins are secreted by ameloblasts as a thin matrix layer that covers the cap enameloid after enameloid maturation.     

Electron-microscope study of the dentine-enamel junction of kangaroo (Macropus giganteus) teeth using selected-area argon-ion-beam thinning

Summary Transmission electron microscopy of selected-area argon-ion-beam thinned kangaroo (Macropus giganteus) enamel revealed a complex ultrastructure in the region of the dentine-enamel junction (DEJ). Characteristic features were multiple branching of dentinal tubules, rejoining of enamel tubules, elongated defects, extended protrusions of dentine into enamel, two types (A and B) of hypomineralized enamel and a continuity between dentinal and enamel tubules. In the intertubular regions of the DEJ a complex intermingling of finer enamel and dentine crystals, similar to that found in human enamel, was observed. The varicosities observed in the light microscope were a combined optical effect caused by the hypomineralized (type A) enamel and the branching and rejoining of the enamel tubules.     

Rat <Emphasis Type="Italic">wct</Emphasis> mutation prevents differentiation of maturation-stage ameloblasts resulting in hypo-mineralization in incisor teeth

A recent study provided genetic and morphological evidence that rat autosomal-recessive mutation, whitish chalk-like teeth (wct), induced tooth enamel defects resembling those of human amelogenesis imperfecta (AI). The wct locus maps to a specific interval of rat chromosome 14 corresponding to human chromosome 4q21 where the ameloblastin and enamelin genes exist, although these genes are not included in the wct locus. The effect of the wct gene mutation on the enamel matrix synthesis and calcification remains to be elucidated. This study clarifies how the wct gene mutation influences the synthesis of enamel matrix and its calcification by immunocytochemistry for amelogenin, ameloblastin and enamelin, and by electron probe micro-analysis (EPMA). The immunoreactivity for enamel proteins such as amelogenin, ameloblastin, and enamelin in the ameloblasts in the homozygous teeth was the same as that in the heterozygous teeth from secretory to transitional stages, although the homozygous ameloblasts became detached from the enamel matrix in the transitional stage. The flattened ameloblasts in the maturation stage of the homozygous samples contained enamel proteins in their cytoplasm. Thus, the wct mutation was found to prevent the morphological transition of ameloblasts from secretory to maturation stages without disturbing the synthesis of enamel matrix proteins, resulting in the hypo-mineralization of incisor enamel and cyst formation between the enamel organ and matrix. This mutation also prevents the transfer of iron into the enamel.     

True enamel matrix of the newt,Triturus pyrrhogaster,contains no sulfated glycoconjugates

Summary The ultrastructural distibution and histochemical properties of sulfated glycoconjugates were investigated in the developing enamel of the adult newt, Triturus pyrrhogaster, by use of the high-iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining and enzymatic digestion methods. Development and ultrastructure of the enamel were also studied. After deposition of the mantle dentin matrix to a certain thickness, the first enamel matrix, globular in shape, appeared in juxtaposition to the dental basement membrane and tended to be intermixed with the previously deposited dentin matrix. Subsequently, enamel matrix was deposited outside (ameloblastic side) of the dental basal lamina and formed a true enamel layer. Thus, developing enamel of the newt consists of two layers: (1) an inner layer made up of a dentin-enamel mixed matrix and (2) an outer layer composed of only true enamel matrix. HID-TCH-SP precipitates resulting from the abovementioned studies were found in the mixed matrix and were identified as chondroitin sulfates; in contrast, the true enamel matrix contained no sulfated glycoconjugates.     

Enamel microstructure in lemuridae (mammalia,primates): Assessment of variability

This study describes the molar enamel microstructure of seven lemurid primates: Hapalemur griseus, Varecia variegata, Lemur catta, Lemur macaco, Lemur fulvus rufus, Lemur fulvus fulvus, and Lemur fulvus albifrons. Contrary to earlier accounts, which reported little or no prism decussation in lemurid enamel, both Lemur and Varecia molars contain a prominent inner layer of decussating prisms (Hunter-Schreger bands), in addition to an outer radial prism layer, and a thin, nonprismatic enamel surface layer. In contrast, Hapalemur enamel consists entirely of radial and, near the surface, nonprismatic enamel. In addition, for all species, prism packing patterns differ according to depth from the tooth surface, and for all species but Varecia (which also has the thinnest enamel of any lemurid), average prism area increases from the enamel-dentine junction to the surface; this may be a developmental solution to the problem of accommodating a larger outer surface area with enamel deposited from a fixed number of cells. Finally, contradicting some previous reports, Pattern 1 prisms predominate only in the most superficial prismatic enamel. In the deeper enamel, prism cross-sections include both closed (Pattern 1) and arc-shaped (Pattern 2 or, most commonly, Pattern 3). This sequence of depth-related pattern change is repeated in all taxa. It should also be emphasized that all taxa can exhibit all three prism patterns in their mature enamel. The high degree of quantitative and qualitative variation in prism size, shape, and packing suggests that these features should be used cautiously in phylogenetic studies. Hapalemur is distinguished from the other lemurids by unique, medially constricted or rectangular prism cross-sections at an intermediate depth and the absence of prism decussation, but, without further assessment of character polarity, these differences do not clarify lemurid phylogenetic relations. Some characters of enamel microstructure may represent synapomorphies of Lemuridae, or of clades within Lemuridae, but homoplasy is likely to be common. Homoplasy of enamel characters may reflect functional constraints. © 1994 Wiley-Liss, Inc.     

Enamel Structure of Cuvieronius hyodon (Proboscidea, Gomphotheriidae) with a Discussion on Enamel Evolution in Elephantoids

Dental material of the South American elephantoid Cuvieronius hyodon from the Late Pleistocene of the Tarija Basin, Bolivia was sampled for a comprehensive analysis of the microstructure of the enamel. To examine variability at the dentition level, enamel samples of the upper incisor, second deciduous premolar, and molars were sectioned. The incisor and cheek teeth enamel is compared to that of other proboscideans in order to reveal phylogenetically and functional informative features useful to reconstruct the evolution of elephantoid enamel. Studies of the adaptations and evolution of proboscidean enamel have focused so far on molars. Nevertheless, given the possibility of an independent evolution of the enamel at different tooth positions, the variation of the enamel throughout the dentition needs to be taken into consideration when using enamel microstructural characters to infer proboscidean diversity and phylogeny. The results obtained from this study demonstrate the generality, among elephantoids, of the basic microstructural features of Cuvieronius hyodon enamel, allowing the characterization of the Elephantoid Enamel (EE). The differentiation between incisor and molar enamel seen in elephantoids is shown to represent a primitive elephantiform trait, as it also occurs in Phiomia. The three-layered enamel of the cheek teeth appears as the sole synapomorphy of the Elephantoidea, though the character might be homoplastic within the Proboscidea. Characters of the prisms cross-section might be used, on the other hand, to define less inclusive clades within the Elephantoidea.     

Unique differentiation of radial enamel in Arsinoitherium (Embrithopoda,Tethytheria)

The enamel microstructure in molars of Arsinoitherium is reinvestigated and a new modification of radial enamel (RE), ‘arsinoitheriid radial enamel (ARE)’, is defined. It is characterised by alternating stripes with different organisation of the interprismatic matrix but no prism decussation. Recognition of this new subtype leads to a reinterpretation of structure previously identified as modified radial enamel and of Hunter–Schreger bands in Arsinoitherium. The newly differentiated ARE of Arsinoitherium is more derived in relation to corresponding microstructures of Palaeoamasia and Crivadiatherium. A careful reinvestigation of RE in other Paenungulata will be required to provide additional data bearing on phylogenetic reconstruction. The enamel of Phenacolophus argues against inclusion of this genus in the Embrithopoda.     

The enamel microstructure ofAnchitheriomys (Rodentia, Mammalia) in comparison with that of other beavers and of porcupines

In the enamel microstructure of the incisors, porcupines (Hystricidae) are characterized by multiserial Hunter-Schreger bands (HSB) while beavers (Castoridae) basically have uniserial HSB. In addition, two groups can be differentiated among beavers based on enamel microstructure.Palaeocastor, Steneofiber andCastor show rather typical uniserial HSB. OnlyCastor shows a slight tendency of fusion of the bands. In contrast,Trogontherium andCastoroides are characterized by the regular occurrence of fused bands.Anchitheriomys, having uniserial HSB and a high frequency of fused bands fits well with theTrogontherium group, proving the castorid nature ofAnchitheriomys. Furthermore, the enamel differentiation observed among the Castoridae thus provides an important character for further systematic subdivision of the beavers. In the enamel of the cheek teeth, porcupines can be told apart from beavers as well.Anchitheriomys shows a two-layered schmelzmuster with inner radial enamel like other beavers. Porcupines have thick HSB throughout the enamel layer. This character in the cheek teeth facilitates the separation of isolated beaver teeth from porcupine teeth.     

Fine structure of the dental enamel in the Family Callitrichidae (Ceboidea,primates)

The fine structure of the dental enamel was intensively examined in the Family Callitrichidae.Leontopithecus rosalia has the nonserial pattern, butCallimico goeldii andSaguinus midas appear more or less modified from the typical nonserial pattern asSaimiri sciureus has (see Figs. 1 & 5). On the other hand,Callithrix jacchus andCebuella pygmaea attain to the primitive stage of the multiserial pattern (see Figs. 4 & 6). So far as the structural patterns of the dental enamel are concerned, a serial trend is confirmed in the Callitrichidae; extending from the genusLeontopithecus through the generaCallimico andSaguinus to the generaCallithrix andCebuella. The genusSaguinus shows extremely wide interspecific variation in the fine structure of the dental enamel, ranging fromS. leucopus with the most primitive features toS. bicolor with the most advanced (see Figs. 2 & 3). The rows of enamel prisms are slightly twisted mesiodistally in the former species, but strongly folded in the latter. This reflects transitional stages from the nonserial pattern to the multiserial. As a whole, however, the genusSaguinus is still regarded as nonserial. Thus, there is a slight, but important gap between the genusSaguinus andCallithrix, as shown by the decussation of clusters (see Fig. 5), as well as by other characters.     

Microstructural defects and enamel hypoplasia in teeth of Toxodon Owen, 1837 from the Pleistocene of Southern Brazil

Enamel hypoplasia is characterized by reduction in the enamel thickness, resulting from a disruption of ameloblast activity due to systemic physiological stress. The euhypsodont teeth of Toxodon, a notoungulate from the Pleistocene of South America, often exhibit signs of enamel hypoplasia, in the form of continuous grooves or a series of pits where the enamel is thinner than in normal areas. These defects alternate with areas of normal enamel, and sometimes more than one form of enamel hypoplasia is present on the same tooth. This study analysed teeth of Toxodon from the Pleistocene Touro Passo Formation and the coastal plain of State of Rio Grande do Sul, Southern Brazil. Six types of enamel hypoplasia were observed. Upper teeth present mainly superficial grooves on the buccal surface, and the defects are less severe than those observed in the lower teeth. In the lower incisors, deep grooves with mesiodistal rows of pits were observed, showing clearly cyclical changes, which to a lesser degree, exist in all teeth. These changes are likely related to the continuous growth of euhypsodont teeth. Seven specimens were analysed under scanning electron microscopy and optical microscopy, which showed the occurrence of microstructural changes associated with the macroscopic enamel defects. Enamel underlying in the vicinity of hypoplastic defects was aprismatic and associated with prominent pathologic striae. These pathological findings might indicate that toxodonts were exposed to some stressing conditions or that their teeth were more easily abraded due to a change in diet items, related to shifting climatic conditions.     

Molar enamel thickness in European Miocene and extant hominoidea

Based on a roentgenographic analysis, the molar enamel of certain European Miocene dryopithecines is absolutely thick (r=1.03–1.30 mm in thickness); the molar enamel of certain European pliopithecines is thin (r=0.32–0.82 mm thick). The rank order for enamel thickness in extant hominoids (from thickest to thinnest) is confirmed to beHomo, Pongo, Gorilla, Pan, andHylobates. There is a great deal of enamel thickness variability within the great ape sample. Extant analogues suggest that dryopithecines were probably adapted to a frugivorous/gramnivorous dietary regimen, while pliopithecines were probably better suited to folivory.     

The effect of rate of eruption on periodontal ligament glycosylaminoglycan content and enamel formation in the rat incisor

The rate of eruption of rat mandibular incisors was either increased by cutting one tooth out of occlusion or eliminated by means of pinning. The effects of such changes in eruption rate on the sulphated glycosylaminoglycan content of the periodontal ligaments was analysed. The length of the enamel secretory zone and the composition of the developing enamel matrix protein was also compared. Sulphated glycosylaminoglycan content of the periodontal ligament increased fourfold (P<0.001) during accelerated eruption but decreased to a corresponding extent (P<0.001) in the absence of eruption, when compared with controls. The length of the enamel secretory zone was also significantly reduced in the immobilised teeth, although the protein content was similar compared with controls. The results demonstrate the differential response to varied eruption rates of the periodontal ligament and enamel, particularly in respect of the extracellular matrix. The data are consistent with the view that the ground substance of the periodontal ligament plays a role in the generation of the eruptive force.     

Rat wct mutation induces a hypo-mineralization form of amelogenesis imperfecta and cyst formation in molar teeth

Our previous findings have demonstrated that the rat autosomal-recessive mutation, whitish chalk-like teeth (wct), induces enamel defects resembling those of human amelogenesis imperfecta (AI) in continuously growing incisor teeth. The present study clarifies the effect of the wct mutation on the morphogenesis and calcification of rat molar teeth. Formalin-fixed maxillae obtained from animals aged 4-30 days were examined by electron probe micro-analysis (EPMA) and by immunocytochemistry for amelogenin, ameloblastin, and enamelin. There were no distinct differences in the calcium and phosphorous contents and the amount of enamel between homozygous mutant and wild-type teeth during postnatal days 4–11. Although the mineral density in the enamel matrix considerably increased in the wild-type teeth until day 15, no changes occurred in mutant teeth during days 11–30. The immunoreactivity for enamel proteins in the secretory-stage ameloblasts in mutant teeth was similar to that in the wild-type teeth, and subsequently mutant maturation-stage ameloblasts became detached from the enamel surface, resulting in odontogenic cyst formation between the enamel organ and matrix until day 7 and the expansion of the cyst around the whole tooth crown on day 15. On day 30, the erupted mutant teeth presented morphological changes such as enamel destruction and tertiary dentin formation in addition to low mineral density in the enamel. Thus, the wct mutation prevents mineral transport without disturbing the synthesis of enamel proteins in molar teeth because of the absence of maturation-stage ameloblasts, in addition to the occurrence of odontogenic cysts. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. This work was supported in part by KAKENHI (B) (no. 16390523 to H.O.) and KAKENHI (C) (no. 18592002 to T.U.) from MEXT, Japan.     

Finite element analysis of stress distribution in intact and porcelain veneer restored teeth

The aim of this study was to investigate the stress distribution generated in a veneer restoration of an upper central incisor compared to intact teeth using the finite element analysis after applying a lingual buccal loading at the incisal edge. Methods: Two models were developed: one model contained enamel, dentine, cementum, periodontal ligament, cortical and trabecullar bones, and the other model was a veneer restoration; both models were developed using MSC/Nastran software (MacNeal-Schwendler Corporation, Los Angeles, CA, USA) as the pre- and post-processor. A 10-N load was applied at the incisal edge from the lingual to the buccal side to simulate oral conditions in this area (protrusion). Results: Von Mises stresses were then analysed for three different regions: A-B (enamel elements under the veneer or second enamel layer), A′-B′ (buccal enamel and/or veneer element layer) and C-D (lingual enamel elements layer). A higher stress mode was observed for both models at the lingual cervical region. Conclusions: The presence of a veneer restoration on the incisors is a good alternative to mimic the behaviour of enamel under protrusion loading conditions. The use of veneers to replace enamel during rehabilitations is recommended.     

In situ expression of the mitochondrial ATPase6 gene in the developing tooth germ of the mouse lower first molar

We previously performed cDNA subtraction between the mouse mandibles on embryonic day 10.5 (E10.5) in the pre-initiation stage of the odontogenesis and E12.0 in the late initiation stage to identify genes expressed at its beginning. Adenosine triphosphate synthase subunit a (Atpase6) is one of the highly expressed genes in the E12.0 mandible including tooth germs. In situ hybridization was conducted using the mouse mandibular first molar from E10.5 to E18.0 to determine the precise expression patterns of Atpase6 mRNA in the developing tooth germ. Atpase6 mRNA was strongly expressed in the presumptive dental epithelium and the underlying mesenchyme at E10.5, and in the thickened dental epithelium at E12.0 and E13.0. Strong in situ signals were observed in the epithelium at E14.0, and in the enamel organ excluded the area of the primary enamel knot at E15.0. Atpase6 was strongly expressed in the inner enamel epithelium, the adjacent stratum intermedium, and the outer enamel epithelium in the cervical loops from E16.0 to E18.0. In addition, strong Atpase6 signals were coincidently demonstrated in various developing cranio-facial organs. These results suggest that Atpase6 participates in the high energy-utilizing functions of the cells related to the initiation and the development of the tooth germ as well as those of the other cranio-facial organs.