Effect of microstructure upon elastic behaviour of human tooth enamel

Tooth enamel is the stiffest tissue in the human body with a well-organized microstructure. Developmental diseases, such as enamel hypomineralisation, have been reported to cause marked reduction in the elastic modulus of enamel and consequently impair dental function. We produce evidence, using site-specific transmission electron microscopy (TEM), of difference in microstructure between sound and hypomineralised enamel. Built upon that, we develop a mechanical model to explore the relationship of the elastic modulus of the mineral–protein composite structure of enamel with the thickness of protein layers and the direction of mechanical loading. We conclude that when subject to complex mechanical loading conditions, sound enamel exhibits consistently high stiffness, which is essential for dental function. A marked decrease in stiffness of hypomineralised enamel is caused primarily by an increase in the thickness of protein layers between apatite crystals and to a lesser extent by an increase in the effective crystal orientation angle.     

Dental wear,wear rate,and dental disease in the African apes

The African apes possess thinner enamel than do other hominoids, and a certain amount of dentin exposure may be advantageous in the processing of tough diets eaten by Gorilla. Dental wear (attrition plus abrasion) that erodes the enamel exposes the underlying dentin and creates additional cutting edges at the dentin‐enamel junction. Hypothetically, efficiency of food processing increases with junction formation until an optimal amount is reached, but excessive wear hinders efficient food processing and may lead to sickness, reduced fecundity, and death. Occlusal surfaces of molars and incisors in three populations each of Gorilla and Pan were videotaped and digitized. The quantity of incisal and molar occlusal dental wear and the lengths of dentin–enamel junctions were measured in 220 adult and 31 juvenile gorilla and chimpanzee skulls. Rates of dental wear were calculated in juveniles by scoring the degree of wear between adjacent molars M1 and M2. Differences were compared by principal (major) axis analysis. ANOVAs compared means of wear amounts. Pearson correlation coefficients were calculated to compare the relationship between molar wear and incidence of dental disease. Results indicate that quantities of wear are significantly greater in permanent incisors and molars and juvenile molars of gorillas compared to chimpanzees. The lengths of dentin–enamel junctions were predominantly suboptimal. Western lowland gorillas have the highest quantities of wear and the most molars with suboptimal wear. The highest rates of wear are seen in Pan paniscus and Pan t. troglodytes, and the lowest rates are found in P.t. schweinfurthii and G. g. graueri. Among gorillas, G. b. beringei have the highest rates but low amounts of wear. Coefficients between wear and dental disease were low, but significant when all teeth were combined. Gorilla teeth are durable, and wear does not lead to mechanical senescence in this sample. Am. J. Primatol. 72:481–491, 2010. © 2010 Wiley‐Liss, Inc.     

High-resolution imaging of proteins in human teeth by scanning probe microscopy

High-resolution studies of dental tissues are of considerable interest for biomedical engineering and clinical applications. In this paper, we demonstrate the application of piezoresponse force microscopy (PFM) to nanoscale imaging of internal structure of human teeth by monitoring the local mechanical response to an electrical bias applied via a conductive tip. It is shown that PFM is capable of detecting dissimilar components of dental tissues, namely, proteins and calcified matrix, which have resembling morphology but different piezoelectric properties. It is demonstrated that collagen fibrils revealed in chemically treated intertubular dentin exhibit high piezoelectric activity and can be visualized in PFM with spatial resolution of 10 nm. Evidence of the presence of protein inclusions of 100-200 nm wide and several micrometers long in tooth enamel has been obtained. Furthermore, it is found that the peritubular dentin and intertubular dentin exhibit different piezoelectric behavior suggesting different concentration of collagen fibrils. The obtained results demonstrate a high potential of PFM in providing an additional insight into the structure of dental tissues. It is suggested that the PFM approach can be used to study the structure of a wide range of biological materials by monitoring their electromechanical behavior at the nanoscale.     

Tooth development in a scincid lizard, Chalcides viridanus (Squamata), with particular attention to enamel formation

Comparative analysis of tooth development in the main vertebrate lineages is needed to determine the various evolutionary routes leading to current dentition in living vertebrates. We have used light, scanning and transmission electron microscopy to study tooth morphology and the main stages of tooth development in the scincid lizard, Chalcides viridanus, viz., from late embryos to 6-year-old specimens of a laboratory-bred colony, and from early initiation stages to complete differentiation and attachment, including resorption and enamel formation. In C. viridanus, all teeth of a jaw have a similar morphology but tooth shape, size and orientation change during ontogeny, with a constant number of tooth positions. Tooth morphology changes from a simple smooth cone in the late embryo to the typical adult aspect of two cusps and several ridges via successive tooth replacement at every position. First-generation teeth are initiated by interaction between the oral epithelium and subjacent mesenchyme. The dental lamina of these teeth directly branches from the basal layer of the oral epithelium. On replacement-tooth initiation, the dental lamina spreads from the enamel organ of the previous tooth. The epithelial cell population, at the dental lamina extremity and near the bone support surface, proliferates and differentiates into the enamel organ, the inner (IDE) and outer dental epithelium being separated by stellate reticulum. IDE differentiates into ameloblasts, which produce enamel matrix components. In the region facing differentiating IDE, mesenchymal cells differentiate into dental papilla and give rise to odontoblasts, which first deposit a layer of predentin matrix. The first elements of the enamel matrix are then synthesised by ameloblasts. Matrix mineralisation starts in the upper region of the tooth (dentin then enamel). Enamel maturation begins once the enamel matrix layer is complete. Concomitantly, dental matrices are deposited towards the base of the dentin cone. Maturation of the enamel matrix progresses from top to base; dentin mineralisation proceeds centripetally from the dentin–enamel junction towards the pulp cavity. Tooth attachment is pleurodont and tooth replacement occurs from the lingual side from which the dentin cone of the functional teeth is resorbed. Resorption starts from a deeper region in adults than in juveniles. Our results lead us to conclude that tooth morphogenesis and differentiation in this lizard are similar to those described for mammalian teeth. However, Tomes processes and enamel prisms are absent.     

牙体预备对高嵌体修复下颌第一磨牙力学影响的初步研究

目的：通过有限元法,探讨高嵌体修复根管治疗后的下颌第一磨牙后,牙体预备形式对牙体组织受力的影响,为临床提供力学理论依据。方法：模拟右侧下颌第一磨牙的三维有限元模型,磨牙存在合面I类洞缺损,根管治疗后采用高嵌体修复,分别设计三种牙体预备形式,即覆盖全部牙尖、覆盖全部功能尖和覆盖部分功能尖的高嵌体修复,对模型进行加载,观察牙体组织的应力大小及分布。结果：覆盖全部牙尖高嵌体修复时牙釉质出现一个应力集中区,其他两种设计出现两个应力集中区。垂直载荷下,釉质的最大主应力在全部牙尖组分别低于其他两组92．29％和89．76％;舌颊向载荷下比其他两组分别降低80．91％和76．53％。三组牙本质的应力集中区趋于一致。垂直载荷下,牙本质的最大主应力在全部牙尖组分别高出其他两组12．92％和14．73％;舌颊向载荷下较其他两组高1．26％和5．08％。结论：从生物力学角度,覆盖全部牙尖的高嵌体更有利于牙体硬组织的受力,可以更好的对牙釉质起到保护作用。     

Size dependent elastic modulus and mechanical resilience of dental enamel

Human tooth enamel exhibits a unique microstructure able to sustain repeated mechanical loading during dental function. Although notable advances have been made towards understanding the mechanical characteristics of enamel, challenges remain in the testing and interpretation of its mechanical properties. For example, enamel was often tested under dry conditions, significantly different from its native environment. In addition, constant load, rather than indentation depth, has been used when mapping the mechanical properties of enamel. In this work, tooth specimens are prepared under hydrated conditions and their stiffnesses are measured by depth control across the thickness of enamel. Crystal arrangement is postulated, among other factors, to be responsible for the size dependent indentation modulus of enamel. Supported by a simple structure model, effective crystal orientation angle is calculated and found to facilitate shear sliding in enamel under mechanical contact. In doing so, the stress build-up is eased and structural integrity is maintained.     

Changes in the matrix proteins, fibronectin and collagen, during differentiation of mouse tooth germ.

The distribution of the matrix protein fibronectin was studied by indirect immunofluorescence in differentiating mouse molars from bud stage to the stage of dentin and enamel secretion, and compared to that of collagenous proteins procollagen type III and collagen type I. Fibronectin was seen in mesenchymal tissue, basement membranes, and predentin. The dental mesenchyme lost fibronectin staining when differentiating into odontoblasts. Fibronectin was not detected in mineralized dentin. Epithelial tissues were negative except for the stellate reticulum within the enamel organ. Particularly intense staining was seen at the epithelio-mesenchymal interface between the dental epithelium and mesenchyme. Fibronectin may here be involved in anchorage of the mesenchymal cells during their differentiation into odontoblasts. Procollagen type III was lost from the dental mesenchyme during odontoblast differentiation but reappeared with advancing vascularization of the dental papilla. Similarly, procollagen type III present in the dental basement membrane during the bud and cap stages disappeared from the cuspal area along with odontoblast differentiation. Weak staining was seen in predentin but not in mineralized dentin. The staining with anti-collagen type I antibodies was weak in dental mesenchyme but intense in predentin as well as in mineralized dentin.     

Nanoindentation study of human premolars subjected to bleaching agent

Bleaching of teeth is gaining popularity due to cosmetic reasons. However, the effect it has on teeth is still largely unknown. This paper seeks to evaluate the effect of a bleaching agent, 30% hydrogen peroxide, on the nanomechanical properties of dentin and enamel using the nanoindentation technique. The Young's modulus and hardness obtained from nanoindentation before and after bleaching were compared. Five newly extracted human premolars were used. Nanoindentation was first done on the sliced enamel and dentin regions to determine their mechanical properties. One batch of samples was kept in Hank's balanced salt solution as control while the other was bleached in 30% hydrogen peroxide for 24h. The same number of nanoindentations was then done near the previously indented regions for both the control and bleached samples and the results compared. Using paired sample t-tests with alpha=0.05, it was found that there were no significant differences in both the Young's modulus and hardness of dentin and enamel kept in control. However, the mechanical properties of the bleached dentin were significantly decreased. For intertubular dentin, the mean hardness decreased by 29-55% and the mean Young's modulus decreased by 19-43%. For enamel, the mean hardness decreased by 13-32% while the mean Young's modulus decreased by 18-32%. The exact mechanism by which hydrogen peroxide affects the dentin and enamel has yet to be fully elucidated. However, it is observed to have an undermining effect on the nanomechanical properties of teeth.     

Synchrotron microComputed Tomography of the mature bovine dentinoenamel junction

The mature dentinoenamel junction (DEJ) is viewed by some investigators and the current authors, not as a fossilized, sharp transition between enamel and dentin, but as a relatively broad structural transition zone including the mantle dentin and the inner aprismatic enamel. In this study, the DEJ structure in bovine incisors was studied with synchrotron microComputed Tomography (microCT) using small cubes cut parallel to the tooth surface. The reconstructions revealed a zone of highly variable punctate contrast between bulk dentin and enamel; the mean linear attenuation coefficients and their standard deviations demonstrated that this zone averaged less mineral than dentin or enamel but had more highly variable structure than either. The region with the punctuate contrast is, therefore, the mantle dentin. The thickness of the mantle dentin seen in a typical data set was about 30 microm, and the mantle dentin-enamel interface deviated +/-15 microm from the average plane over a distance of 520 microm. In the highest resolution data ( approximately 1.5 microm isotropic voxels, volume elements), tubules in the dentin could be discerned in the vicinity of the DEJ. Contrast sensitivity was high enough to detect differences in mineral content between near-surface and near-DEJ volumes of the enamel. Reconstructions before and after two cubes were compressed to failure revealed cracks formed only in the enamel and did not propagate across the mantle dentin, regardless of whether loading was parallel to or perpendicular to the DEJ.     

FAM20C Plays an Essential Role in the Formation of Murine Teeth

FAM20C is highly expressed in bone and tooth. Previously, we showed that Fam20C conditional knock-out (KO) mice manifest hypophosphatemic rickets, which highlights the crucial roles of this molecule in promoting bone formation and mediating phosphate homeostasis. In this study, we characterized the dentin, enamel, and cementum of Sox2-Cre-mediated Fam20C KO mice. The KO mice exhibited small malformed teeth, severe enamel defects, very thin dentin, less cementum than normal, and overall hypomineralization in the dental mineralized tissues. In situ hybridization and immunohistochemistry analyses revealed remarkable down-regulation of dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein in odontoblasts, along with a sharply reduced expression of ameloblastin and amelotin in ameloblasts. Collectively, these data indicate that FAM20C is essential to the differentiation and mineralization of dental tissues through the regulation of molecules critical to the differentiation of tooth-formative cells.     

Mucopolysaccharides in odontogenesis

Summary Localization of sulfomucopolysaccharides in developing teeth of Swiss albino mice was detected by S³⁵ autoradiography and histochemistry.A positive correlation was found to exist between autoradiographic and histochemical data with regard to the localization of sulfomucopolysaccharides. Autoradiography, however, revealed some sites of localization which were not detectable by histochemistry, namely, the odontoblasts and stratum intermedium.Fetuses which received the isotope via maternal injection at the cap stage of tooth development and were sacrificed after 2 hours of isotope action displayed rapid incorporation of the isotope in the components of the dental papilla. In the enamel organ, however, only moderate activity was recorded. When the time interval between injection and sacrifice of the experimental animals was increased to 20 hours, intense activity was observed in the enamel organ. With progressively longer intervals between injection and sacrifice, S³⁵ was demonstrable first in odontoblasts and later in the predentin. This occurred as a band or active zone which migrated toward the dentino-enamel junction. With the increasing intervals between injection and sacrifice, first the odontoblasts were active, then predentin was active while the odontoblasts became reduced in activity, after which the dentin matrix gained activity while the predentin decreased somewhat in activity. This pattern is consistent with appositional growth. A linear band of activity was not observed in the enamel matrix; rather, the activity was present as a diffuse stippling over a relatively large area of the matrix. The sulfomucopolysaccharide which existed in dentin matrix was postulated to have originated from the cells of both the odontoblastic layer and the dental papilla.Supported by PHS Grant No 2800-02, Tooth Germ Development, National Institute of Dental Research, National Institutes of Health.     

Comparative morphology of incisor enamel and dentin in humans and fat dormice (Glis glis)

The structure of teeth in all living beings is genetically predetermined, although it can change under external physiological and pathological factors. The author's hypothesis was to indicate evolutional shifts resulting from genetic, functional and other differences. A comparative study about certain characteristics of incisors in humans and myomorpha, the fat dormouse (Glis glis) being their representative as well, comprised measurements of enamel and dentin thickness in individual incisor segments, evaluation of external enamel index, and also assessment of histological structure of enamel and dentin. The study results involving dormice showed the enamel to be thicker in lower than in the upper teeth, quite contrary to enamel thickness in humans. In the upper incisors in dormice the enamel is the thickest in the medial layer of the crown, and in the cervical portion of the crown in the lower incisors. The thickness of dentin in dormice is greater in the oral than in the vestibular side. These findings significantly differ from those reported in reference literature, but they are based on the function of teeth in dormice. Histological characteristics of hard dental tissues in dormice are similar to those in humans, with exception of uniserial structure of enamel and appearance of dentinoenamel junction.     

Dental Phenotype in Jalili Syndrome Due to a c.1312 dupC Homozygous Mutation in the CNNM4 Gene

Jalili syndrome denotes a recessively inherited combination of an eye disease (cone-rod dystrophy) and a dental disorder (amelogenesis imperfecta), which is caused by mutations in the CNNM4 gene. Whereas the ophthalmic consequences of these mutations have been studied comprehensively, the dental phenotype has obtained less attention. A defective transport of magnesium ions by the photoreceptors of the retina is assumed to account for the progressive visual impairment. Since magnesium is also incorporated in the mineral of dental hard tissues, we hypothesized that magnesium concentrations in defective enamel resulting from mutations in CNNM4 would be abnormal, if a similar deficiency of magnesium transport also accounted for the amelogenesis imperfecta. Thus, a detailed analysis of the dental hard tissues was performed in two boys of Kosovan origin affected by Jalili syndrome. Retinal dystrophy of the patients was diagnosed by a comprehensive eye examination and full-field electroretinography. A mutational analysis revealed a c.1312 dupC homozygous mutation in CNNM4, a genetic defect which had already been identified in other Kosovan families and putatively results in loss-of-function of the protein. The evaluation of six primary teeth using light and scanning electron microscopy as well as energy-dispersive X-ray spectroscopy showed that dental enamel was thin and deficient in mineral, suggesting a hypoplastic/hypomineralized type of amelogenesis imperfecta. The reduced mineral density of enamel was accompanied by decreased amounts of calcium, but significantly elevated levels of magnesium. In dentin, however, a similar mineral deficiency was associated with reduced magnesium and normal calcium levels. It is concluded that the c.1312 dupC mutation of CNNM4 results in mineralization defects of both enamel and dentin, which are associated with significantly abnormal magnesium concentrations. Thus, we could not disprove the hypothesis that a disrupted magnesium transport is involved in the development of the dental abnormalities observed in Jalili syndrome.     

Nanoindentation of lemur enamel: an ecological investigation of mechanical property variations within and between sympatric species

The common morphological metrics of size, shape, and enamel thickness of teeth are believed to reflect the functional requirements of a primate's diet. However, the mechanical and material properties of enamel also contribute to tooth function, yet are rarely studied. Substantial wear and tooth loss previously documented in Lemur catta at the Beza Mahafaly Special Reserve suggests that their dental morphology, structure, and possibly their enamel are not adapted for their current fallback food (the mechanically challenging tamarind fruit). In this study, we investigate the nanomechanical properties, mineralization, and microstructure of the enamel of three sympatric lemur species to provide insight into their dietary functional adaptations. Mechanical properties measured by nanoindentation were compared to measurements of mineral content, prism orientation, prism size, and enamel thickness using electron microscopy. Mechanical properties of all species were similar near the enamel dentin junction and variations correlated with changes in microstructure (e.g., prism size) and mineral content. Severe wear and microcracking within L. catta's enamel were associated with up to a 43% reduction in nanomechanical properties in regions of cracking versus intact enamel. The mechanical and material properties of L. catta's enamel are similar to those of sympatric folivores and suggest that they are not uniquely mechanically adapted to consume the physically challenging tamarind fruit. An understanding of the material and mechanical properties of enamel is required to fully elucidate the functional and ecological adaptations of primate teeth.     

Ontogenetic variations in Sr/Ca and Ba/Ca ratios of dental bioapatites from Bos taurus and Odocoileus virginianus

Sr/Ca and Ba/Ca ratios of bone are commonly used as biochemical indicators of trophic level in modern and fossil mammals. Concerns over the effects of diagenesis on Sr/Ca and Ba/Ca ratios of bone led archaeologists and paleontologists to favor tooth enamel, which is less prone to alteration. Sr/Ca and Ba/Ca ratios of bone, enamel, and dentin from three farm-raised steers (Bos taurus) and five wild white-tailed deer (Odocoileus virginianus) from central Missouri were compared. Our results show that changes in diet, discrimination, and growth rate during ontogeny can lead to significant differences in Sr/Ca and Ba/Ca ratios of different bioapatite types as well as significant differences within the same bioapatite forming at different times. Early- and late-forming tooth enamel can have significant differences in Sr/Ca and Ba/Ca ratios equivalent to almost one full trophic step. Although differences between early- and late-forming dentin are typically not significant, dentin Sr/Ca and Ba/Ca ratios are significantly greater than enamel values. This difference in Sr/Ca or Ba/Ca ratios between enamel and dentin from the same tooth can be greater than one full trophic step. These results have profound implications for the use of dental bioapatites in trophic level reconstructions. They highlight the importance of consistency in bioapatite selection, tooth selection, and relative location of sampling within the enamel cap. Furthermore, this expected difference in Sr/Ca and Ba/Ca ratios could be used as another means of checking for diagenetic alteration in ancient samples.     

Stress analysis of irradiated human tooth enamel using finite element methods

The objectives of this project were to use finite element methods to determine how changes in the elastic modulus due to oral cancer therapeutic radiation alter the distribution of mechanical stresses in teeth and to determine if observed failures in irradiated teeth correlate with changes in mechanical stresses. A thin slice section finite element (FE) model was constructed from micro CT sections of a molar tooth using MIMICS and 3-Matic software. This model divides the tooth into three enamel regions, the dentin-enamel junction (DEJ) and dentin. The enamel elastic modulus was determined in each region using nano indentation for three experimental groups namely – control (non-radiated), in vitro irradiated (simulated radiotherapy following tooth extraction) and in vivo irradiated (extracted subsequent to oral cancer patient radiotherapy) teeth. Physiological loads were applied to the tooth models at the buccal and lingual cusp regions for all three groups (control, in vitro and in vivo). The principal tensile stress and the maximum shear stress were used to compare the results from different groups since it has been observed in previous studies that delamination of enamel from the underlying dentin was one of the major reasons for the failure of teeth following therapeutic radiation. From the FE data, we observed an increase in the principal tensile stress within the inner enamel region of in vivo irradiated teeth (9.97 ± 1.32 MPa) as compared to control/non-irradiated teeth (8.44 ± 1.57 MPa). Our model predicts that failure occurs at the inner enamel/DEJ interface due to extremely high tensile and maximum shear stresses in in vivo irradiated teeth which could be a cause of enamel delamination due to radiotherapy.     

Ectopic expression of dentin sialoprotein during amelogenesis hardens bulk enamel

Dentin sialophosphpoprotein (Dspp) is transiently expressed in the early stage of secretory ameloblasts. The secretion of ameloblast-derived Dspp is short-lived, correlates to the establishment of the dentinoenamel junction (DEJ), and is consistent with Dspp having a role in producing the specialized first-formed harder enamel adjacent to the DEJ. Crack diffusion by branching and dissipation within this specialized first-formed enamel close to the DEJ prevents catastrophic interfacial damage and tooth failure. Once Dspp is secreted, it is subjected to proteolytic cleavage that results in two distinct proteins referred to as dentin sialoprotein (Dsp) and dentin phosphoprotein (Dpp). The purpose of this study was to investigate the biological and mechanical contribution of Dsp and Dpp to enamel formation. Transgenic mice were engineered to overexpress either Dsp or Dpp in their enamel organs. The mechanical properties (hardness and toughness) of the mature enamel of transgenic mice were compared with genetically matched and age-matched nontransgenic animals. Dsp and Dpp contributions to enamel formation greatly differed. The inclusion of Dsp in bulk enamel significantly and uniformly increased enamel hardness (20%), whereas the inclusion of Dpp weakened the bulk enamel. Thus, Dsp appears to make a unique contribution to the physical properties of the DEJ. Dsp transgenic animals have been engineered with superior enamel mechanical properties.     

High energy X-ray scattering quantification of in situ-loading-related strain gradients spanning the dentinoenamel junction (DEJ) in bovine tooth specimens

High energy X-ray scattering (80.7 keV photons) at station 1-ID of the Advanced Photon Source quantified internal strains as a function of applied stress in mature bovine tooth. These strains were mapped from dentin through the dentinoenamel junction (DEJ) into enamel as a function of applied compressive stress in two small parallelepiped specimens. One specimen was loaded perpendicular to the DEJ and the second parallel to the DEJ. Internal strains in enamel and dentin increased and, as expected from the relative values of the Young’s modulus, the observed strains were much higher in dentin than in enamel. Large strain gradients were observed across the DEJ, and the data suggest that the mantle dentin-DEJ-aprismatic enamel structure may shield the near-surface volume of the enamel from large strains. In the enamel, drops in internal strain for applied stresses above 40 MPa also suggest that this structure had cracked.     

The dentinoenamel junction in primates

Dental characteristics figure large in primate taxonomy because teeth fossilize commonly and reflect dietary adaptations. The mammalian dentinoenamel junction (DEJ) plays a crucial role throughout odontogenesis in determining the ultimate crown configuration, being the interface between the papilla and the dental cap. The final configuration of the dentin surface reflects the epithelium more closely than does the enamel surface of the crown. Enamel deposition occurs relatively late in calcification, often causing many changes from the pattern residing in the DEJ; the genetic determination of the two surfaces also differs. Comparative study of the DEJ (and its differences from the enamel crown) is a potential adjunct to dentition-based taxonomy and may help to resolve certain cusp homologies and morphogenetic order of appearance. Primate teeth were stripped of enamel after measurement, mapping, and anatomical observations on the original crown. The dentin surfaces thus revealed differ from the enamel surfaces in several respects and shed new light on such dental problem areas as the origin of the hypocone, the affinities of lorisoids and callitrichids, the monophyly of the anthropoid grade, and human affinities.     

The lingual (root analogue) and the labial (crown analogue) mouse incisor dentin promotes ameloblast differentiation

Previous studies have suggested that mouse molar ameloblast differentiation was triggered by the predentin-dentin. Knowing that enamel is absent on the lingual surface of the mouse incisor, the aim of this study was to compare in heterotopic tissue recombinations the behavior of mouse molar inner dental epithelium associated with lingual or labial mouse incisor dentin. It was shown that root-analogue and crown-analogue incisor dentin promotes ameloblast differentiation of competent molar inner dental epithelium.