Spectrally enhanced imaging of occlusal surfaces and artificial shallow enamel erosions with a scanning fiber endoscope

An ultrathin scanning fiber endoscope, originally developed for cancer diagnosis, was used to image dental occlusal surfaces as well as shallow artificially induced enamel erosions from human extracted teeth (n=40). Enhanced image resolution of occlusal surfaces was obtained using a short-wavelength 405-nm illumination laser. In addition, artificial erosions of varying depths were also imaged with 405-, 404-, 532-, and 635-nm illumination lasers. Laser-induced autofluorescence images of the teeth using 405-nm illumination were also obtained. Contrast between sound and eroded enamel was quantitatively computed for each imaging modality. For shallow erosions, the image contrast with respect to sound enamel was greatest for the 405-nm reflected image. It was also determined that the increased contrast was in large part due to volume scattering with a smaller component from surface scattering. Furthermore, images obtained with a shallow penetration depth illumination laser (405 nm) provided the greatest detail of surface enamel topography since the reflected light does not contain contributions from light reflected from greater depths within the enamel tissue. Multilayered Monte Carlo simulations were also performed to confirm the experimental results.     

Expression survey of genes critical for tooth development in the human embryonic tooth germ.

In the developing murine tooth, the expression patterns of numerous regulatory genes have been examined and their roles have begun to be revealed. To unveil the molecular mechanisms that regulate human tooth morphogenesis, we examined the expression patterns of several regulatory genes, including BMP4, FGF8, MSX1, PAX9, PITX2, and SHOX2, and compared them with that found in mice. All of these genes are known to play critical roles in murine tooth development. Our results show that these genes exhibit basically similar expression patterns in the human tooth germ compared with that in the mouse. However, slightly different expression patterns were also observed for some of the genes at certain stages. For example, MSX1 expression was detected in the inner enamel epithelium in addition to the dental mesenchyme at the bell stage of the human tooth. Moreover, FGF8 expression remained in the dental epithelium at the cap stage, while PAX9 and SHOX2 expression was detected in both dental epithelium and mesenchyme of the human tooth germ. Our results indicate that, although slight differences exist in the gene expression patterns, the human and mouse teeth not only share considerable homology in odontogenesis but also use similar underlying molecular networks.     

On the chipping and splitting of teeth

One of the most frequent fracture modes in teeth is chipping. It can lead to deterioration and ultimate loss of tooth function. Chips in enamel can also be used to gain insight into the evolutionary history of extant animal and fossil hominin species. In this study, chipping tests are performed on the surfaces of as-received or flattened human molars using hard indenters. The chips exhibit a characteristic scallop shape, with some influence from tooth curvature as well as from enamel anisotropy and inhomogeneity. Chipping fracture tends to follow easy interprism pathways, but inevitably involves breakage of bundles of mineralized prisms in the last stages of spallation. A simple relation describes how critical loads for chipping scale with distance of the occlusal contact from the specimen edge. Measured loads fall well within the range of biting forces exerted during normal oral function. A transition from chipping to splitting occurs at higher loads for contacts nearer the central axis of the tooth.     

Three-dimensional direction and interrelationship of prisms in cuspal and cervical enamel of dog tooth

The three-dimensional architecture of enamel prisms was examined in cuspal enamel and compared with that in cervical enamel by light and electron microscopy as well as computer-assisted reconstruction using the developing enamel of several dog teeth. Dog tooth enamel consists of two groups of alternately arranged enamel prisms oriented in opposite sideward directions basically forming thick horizontal rings, partly branching off from the stem. Along a 8–10 enamel prism-wide group, the enamel prisms emerge in parallel tilting uniformly to the same sideward direction. In cervices, groups of enamel prisms are arranged nearly in parallel displaying a regular arrangement of prisms. Approaching the cusp of tooth, the groups of enamel prisms fuse to a concentric cusp-centered arrangement and the prisms exhibit no periodic arrangement as shown in the cervical enamel. It is suggested that the three-dimensional structure of enamel becomes complicated close to the cusp, contributing to the chewing stress of tooth. Anat. Rec. 252:355–368, 1998. © 1998 Wiley-Liss, Inc.     

Modulation of activin/bone morphogenetic protein signaling by follistatin is required for the morphogenesis of mouse molar teeth.

Teeth form as ectodermal appendages, and their morphogenesis is regulated by conserved signaling pathways. The shape of the tooth crown results from growth and folding of inner dental epithelium, and the cusp patterning is regulated by transient signaling centers, the enamel knots. Several signal proteins in the transforming growth factor-beta (TGF beta) superfamily are required for tooth development. Follistatin is an extracellular inhibitor of TGF beta signaling. To investigate the roles of follistatin during tooth development, we analyzed in detail the expression patterns of follistatin, activin beta A, as well as Bmp2, Bmp4, and Bmp7 during tooth morphogenesis. We also examined the tooth phenotypes of follistatin knockout mice and of transgenic mice overexpressing follistatin in the epithelium under the keratin 14 (K14) promoter. The folding of the dental epithelium was aberrant in the molars of follistatin knockout mice, and the cusps were shallow with reduced cell proliferation and lack of anteroposterior polarization. The functions of both primary and secondary enamel knots were apparently disturbed. In K14-follistatin transgenic mice, the molar cusp pattern was also seriously affected (although different from the follistatin knockouts) and the occlusal surfaces of the molars were whorled. Their enamel was prematurely worn. In addition, all of the third molars were missing. Our results indicate that follistatin regulates morphogenesis and shaping of the tooth crown. We propose that finely tuned antagonistic effects between follistatin and TGF beta superfamily signals are critical for enamel knot formation and function, as well as for patterning of tooth cusps.     

Immunohistochemical localization of connexin 43 in the developing tooth germ of rat

Distribution of gap junction protein in maxillary tooth germs of 1-day-old rats was examined by immunohistochemistry, using an affinity-purified antibody specific to residues 360–376 of rat connexin (CX) 43. In 1-day-old rats, the maxillary second molar formed the shape of the cusp, but neither dentine nor enamel was formed between the cells of the dental papilla and the inner enamel epithelium. In the tooth germ, CX 43 was expressed in the cells of the stratum intermedium and the inner enamel epithelium. Labelling in the stratum inter-medium was extensive and showed an increasing gradient from peripheral to cuspal regions. CX 43 detected in the inner enamel epithelium was at cell surfaces facing the interface between the dental papilla and the inner enamel epithelium. The cells of the dental papilla and the inner enamel epithelium began differentiation as odontoblasts and secretory ameloblasts respectively, in the cusps of the first molars, where predentine and dentine were formed but enamel matrix was not secreted. CX 43 was present in the stratum intermedium, inner enamel epithelium, preodontoblasts, odontoblasts and subodontoblasts. The incisors showed the most advanced stage of development, where the enamel matrix and calcified dentine were formed in the labial part of the teeth. The CX 43 epitope was seen in the stratum intermedium, inner enamel epithelium, preameloblasts, preodontoblasts, odontoblasts, and subodontoblasts. Immunolabelling was more extensive in the stratum intermedium and subodontoblasts than in preameloblasts, preodontoblasts, and odontoblasts. The immunolabelling in preameloblasts and preodontoblasts was accumulated at cell surfaces facing the predentine. Further, the labelling in preameloblasts and preodontoblasts disappeared or was reduced at the initiation of enamel matrix secretion and calcification of dentine matrix.The present results suggest that gap junctional cell communication has important roles in tooth development. Further, the extensive CX 43 expression in the stratum intermedium and the subodontoblast layer suggests that gap junctions have an important role in amelogenesis and dentinogenesis.     

Biomechanical Evaluation of Equine Masticatory Action: Position and Curvature of Equine Cheek Teeth and Age‐Related Changes

The equine cheek tooth battery is part of a very dynamic system. The aim of this study was to investigate whether the curvature and position of the teeth are also involved in such dynamical processes. The alveolar crest was labelled with a radiodense marker (48 cadaver heads, 15 skulls) and laterolateral radiographs were taken. Then a geometrical method was elaborated to determine a cheek tooth's curvature and its position by means of specific angles. This method respects the remarkable changes of the equine dentition throughout life by considering two items: (1) the alveolar crest was taken as a constant landmark, (2) the central axis of the curved dental crown was determined by calculation of a linear regression equation. This equation considered several geometrically determined points on the curved dental crown which had been marked in the radiographs. Our study yielded the following results: Mandibular cheek teeth became more curved with age, but their positions (represented by the so‐called mesio‐occlusal angle between tooth and alveolar crest) did not change significantly. In maxillary cheek teeth, however, the mesio‐occlusal angle became larger with age (indication of change of dental position), while their curvature did not change. Even though changes of the dental position were not always statistically significant, they are discussed as being biologically/functionally relevant. The mandibular anticlinal tooth, i.e. the tooth positioned at a mesio‐occlusal angle of about 90°, was not in contact with the maxillary anticlinal tooth. Interestingly, the maxillary anticlinal tooth is known to cause most clinical dental problems. Anat Rec, 291:565–570, 2008. © 2008 Wiley‐Liss, Inc.     

Regulation of enamel hardness by its crystallographic dimensions

Enamel is a composite biomaterial comprising a minor organic matrix (~2%) and a hierarchically organized inorganic ultrastructure (~96-98%). Surprisingly, to date there is no available information in the literature regarding the possible role of the enamel ultrastructure on the nanoscale level in tooth macroscopic properties. Understanding this relationship is of special interest for restorative purposes in dentistry. Accordingly, this study was designed to investigate how enamel nanocrystals regulate its hardness. We performed microindentation analysis on 100 extracted human teeth. The tooth enamel hardness was quantified and correlated with changes in enamel chemical composition and crystallographic dimensions obtained from Fourier transform infrared spectroscopy and X-ray diffraction, respectively. Enamel hardness was not related to the variability in organic content, but was associated with the size of apatite crystals along the c-axis. This association followed the Hall-Petch model for polycrystalline materials, indicating that the optimal size of apatite nanocrystals (larger than the critical size) provides enamel with the greatest hardness, which enables teeth to survive the heavy wear over a human lifetime.     

胎儿牙齿硬组织发育的临床应用解剖

目的：探讨国人胎儿牙齿硬组织发育状况。方法：对正常胎龄10-38周的胎儿上、下颌骨作连续切片,观察乳、恒牙胚及其相关结构发生规律。结果：牙体硬组织形成于14-20周,牙齿的增殖、钙化、萌出同时进行,牙釉质、牙本质厚度随胎龄增长而增加,各乳磨牙牙尖总是近颊尖最先发牛,恒牙板总是位于乳牙胚舌侧。结论：国人牙体硬组织发育成熟较低,牙釉质、牙本质厚度与胎龄呈正相关。     

Tooth development in a model reptile: functional and null generation teeth in the gecko Paroedura picta

This paper describes tooth development in a basal squamate, Paroedura picta. Due to its reproductive strategy, mode of development and position within the reptiles, this gecko represents an excellent model organism for the study of reptile development. Here we document the dental pattern and development of non-functional (null generation) and functional generations of teeth during embryonic development. Tooth development is followed from initiation to cytodifferentiation and ankylosis, as the tooth germs develop from bud, through cap to bell stages. The fate of the single generation of non-functional (null generation) teeth is shown to be variable, with some teeth being expelled from the oral cavity, while others are incorporated into the functional bone and teeth, or are absorbed. Fate appears to depend on the initiation site within the oral cavity, with the first null generation teeth forming before formation of the dental lamina. We show evidence for a stratum intermedium layer in the enamel epithelium of functional teeth and show that the bicuspid shape of the teeth is created by asymmetrical deposition of enamel, and not by folding of the inner dental epithelium as observed in mammals.     

Microbiological and microhardness evaluation of artificial enamel fissures worn intraorally by humans.

Devices simulating occlusal fissures were constructed from dental enamel and bonded to human maxillary molar teeth for 3 weeks. Facilitation of in vivo caries research in humans is the long-range goal for this model system. In the present investigation, microbial compositions of plaque in the fissural space of the model and natural fissural plaque from teeth bearing the devices were compared. Plaque from models constructed from either bovine or human enamel was also compared. In addition, microhardness of the enamel surfaces was examined before and after oral exposure. Plaque in both bovine and human enamel models differed significantly from natural fissural plaque in several microbial categories. Differences appeared to be related to the increased accessibility of the deep fissural contents of the models. For example, levels of the aciduric Streptococcus mutans and Veillonella species were higher in the models, although levels of a salivary inhabitant, Streptococcus salivarius, were lower. Plaque in bovine and human enamel models was similar, and both models showed significant decreases in enamel microhardness after oral exposure for 3 weeks. The model system should be useful in caries research in that carieslike activity was simulated within a relatively short period of time, and the fissural space became colonized by high levels of cariogenic bacteria.     

The Enamel Microstructures of Bovine Mandibular Incisors

Bovine teeth have been considered as an excellent substitute for human teeth for dental research, however, the enamel microstructures of bovine incisors that include arrangements of prisms and interprisms, and their spatial relationships have not been well described. The aim of this study was to investigate the detail enamel microstructures of bovine incisors. Eight bovine mandibular incisors were cut into 77 pieces at eight equal intervals either in the longitudinal direction or in the horizontal direction before each piece had been tangentially cut (parallel to enamel–dentin junction) through the middle of the enamel thickness. All the sectioned surfaces were treated 1 M HCl for 10 sec to expose the prisms and interprisms before observation by scanning electron microscopy. The parallel enamel prisms were located in all the outer enamel, the cervical region and the incisal ridge of the bovine incisors. Most labial inner enamel and the cingulum of lingual inner enamel were composed of the Hunter–Schreger bands with the characteristics of decussating groups of prisms and decussating planes between interprisms and prisms. The interprisms were thicker in the inner enamel than in the outer enamel. Anat Rec, 2012. © 2012 Wiley Periodicals, Inc.     

Exploring metameric variation in human molars: a morphological study using morphometric mapping

Human molars exhibit a type of metameric variation, which is the difference in serially repeated morphology within an organism. Various theories have been proposed to explain how this variation is brought about in the molars. Actualistic data that support the theories, however, are still relatively scarce because of methodological limitations. Here we propose new methods to analyse detailed tooth crown morphologies. We applied morphometric mapping to the enamel–dentine junction of human maxillary molars and examined whether odontogenetic models were adaptable to human maxillary molars. Our results showed that the upper first molar is phenotypically distinct among the maxillary molars. The average shape of the upper first molar is characterized by four well‐defined cusps and precipitous surface relief of the occlusal table. On the other hand, upper third molar is characterized by smooth surface relief of the occlusal table and shows greater shape variation and distinct distribution patterns in morphospace. The upper second molar represents an intermediate state between first and third molar. Size‐related shape variation was investigated by the allometric vector analysis, and it appeared that human maxillary molars tend to converge toward the shape of the upper first molar as the size increases. Differences between the upper first molar and the upper second and third molar can thus be largely explained as an effect of allometry. Collectively, these results indicate that the observed pattern of metameric variation in human molars is consistent with odontogenetic models of molar row structure (inhibitory cascade model) and molar crown morphology (patterning cascade model). This study shows that morphometric mapping is a useful tool to visualize and quantify the morphological features of teeth, which can provide the basis for a better understanding of tooth evolution linking morphology and development.     

Distribution of actin bundles in periodontal ligament of rat lower incisor

The overall distribution of actin bundles in fibroblasts in the periodontal ligament (PDL) was examined in continuously growing incisors of the rat by confocal laser and thin-section electron microscopy to better understand the functional roles of PDL in mechanical support of the tooth and the mechanism of tooth eruption. The PDL around the incisor was divided mainly into two parts: the superior (non-enamel side of the tooth) and inferior (enamel side) parts. In the superior part, elongated fibroblasts ran obliquely near the alveolar bone and shifted direction nearly parallel to the tooth as they approached the tooth. Long and thin bundles of actin filaments were observed running along the long axis of the cells. In the inferior part, flattened cells were densely populated in the narrow space between the enamel epithelium and venous plexus with their long axis parallel to the tooth, and contained relatively thick actin bundles running parallel to the cell axis. Thus, different distribution patterns of fibroblasts and their actin bundles were recognized in different parts of the PDL. These observations suggest that different parts of the PDL of the rat lower incisor may have different functional significances such as force generation of tooth eruption and mechanical resistance to occlusal pressure and continuous growth.     

X-ray microtomographic study of mineral distribution in enamel of mandibular rat incisors

X-ray microtomography was used to study the mineral concentrations in sequential slices of enamel of 5 mandibular incisors which showed an increase from ∼ 1.0 to ∼ 2.7 g cm⁻³ from the apex towards the incisal end. For points at the same distance from the apex, there were differences up to 0.6 g cm⁻³ between the teeth. The change of mean concentrations in the slices with distance could be modelled as (different) saturating exponentials. Under the assumption of a uniform growth rate of a mandibular incisor of 0.6 mm per day and a common time origin for the start of maturation (taken as a mineral concentration of 1 g cm⁻³), the distances were transformed to a common time frame to give a pooled data set. A single saturating exponential could be fitted to this pooled transformed data; this was: C _m = 2.84−1.94exp (−0.18 d ) where Cm is the mean mineral concentration (g cm⁻³) and d the time (days) from the start of maturation. This gives an asymptotic concentration of 2.84 g cm⁻³ towards the incisal end, with a time constant of 7.7 days. The mineral concentration distribution functions were found to be more positively skewed closer to the apex, but more negatively skewed towards the incisal end. The difference between the higher mineral concentration in the outer enamel and the enamel near the amelodentinal junction (ADJ) was ∼ 3%. The direction of maximum increase in concentration from the outer enamel surface to the ADJ meets the boundary of the ADJ at ∼ 80°. Three dimensional surface rendering of isodensity contours showed that the previously described C-shaped pattern of mineralisation is not solely a surface phenomenon, but extends through the depth of the enamel.     

A simple model for enamel fracture from margin cracks

We present results of in situ fracture tests on extracted human molar teeth showing failure by margin cracking. The teeth are mounted into an epoxy base and loaded with a rod indenter capped with a Teflon insert, as representative of food modulus. In situ observations of cracks extending longitudinally upward from the cervical margins are recorded in real time with a video camera. The cracks appear above some threshold and grow steadily within the enamel coat toward the occlusal surface in a configuration reminiscent of channel-like cracks in brittle films. Substantially higher loading is required to delaminate the enamel from the dentin, attesting to the resilience of the tooth structure. A simplistic fracture mechanics analysis is applied to determine the critical load relation for traversal of the margin crack along the full length of the side wall. The capacity of any given tooth to resist failure by margin cracking is predicted to increase with greater enamel thickness and cuspal radius. Implications in relation to dentistry and evolutionary biology are briefly considered.     

Interproximal contact hypoplasia in primary teeth: A new enamel defect with anthropological and clinical relevance

This study reports the prevalence, distribution, and expression of enamel defects in a sample of primary teeth (n = 225) from a prehistoric site in western India (1400–700 BC). Five enamel surfaces of individual, isolated primary teeth were observed for surface defects using a binocular stereomicroscope with variable power of magnification (8–20×). Standards for evaluating dental enamel defects (DDE) recommended by the Fédération Dentaire International (FDI) were employed. Details of defect expression were also recorded, including size, shape, and surface of tooth crown affected. Hypoplastic enamel defects were observed in 28% of teeth, but the distribution and expression of defects was not random. More than 50% of canine teeth had hypoplastic defects (HD); incisors and molar teeth exhibited far fewer HD. The buccal surface of canines was the most commonly affected crown surface. Areas of missing enamel were also common on the mesial and distal surfaces of canines and incisors and on the mesial surface of molar teeth. The high frequency of enamel defects found on interproximal crown surfaces warrants a label, and the name interproximal contact hypoplasia (IPCH) is proposed. Linear enamel hypoplasia (LEH) was absent from this primary dental sample. IPCH is more frequent in mandibular than in maxillary teeth, but no side preference was detected. In canine teeth, buccal hypoplasias (localized hypoplasia of primary canines; LHPC) were not positively correlated with interproximal hypoplastic defects. The etiology of IPCH may involve mesial compaction of developing teeth due to slow longitudinal growth of the jaws. Episodic bone remodeling results in ephemeral fenestrae in the mesial and distal walls of the dental crypt permitting tooth–tooth contact and disruption of amelogenesis. IPCH prevalence decreases across the subsistence transition from sedentary Early Jorwe agriculturalists to seminomadic Late Jorwe hunters and foragers, but the difference is not statistically significant. This may be due to underrepresentation of mandibular teeth in the sample. Am. J. Hum. Biol. 11:718–734, 1999. © 1999 Wiley-Liss, Inc.     

Analysis of fracture and deformation modes in teeth subjected to occlusal loading

An analysis of fracture and deformation modes in tooth enamel subjected to occlusal loading is presented. Several competing modes are identified: deformation by yield beneath the indenter; median cracking from the ensuing plastic zone and analogous radial cracking from the dentin–enamel junction along the load axis; and margin cracking from the cervical enamel–cement junction. The analysis, based on a simple model of tooth geometry, presents relations for the critical loads to initiate these damage modes within the enamel, and to drive ensuing cracks longitudinally around the tooth walls to failure. The relations are explicit in their dependence on characteristic tooth dimensions – enamel thickness and cuspal radius – and on material properties – modulus, hardness, toughness and strength. Provision is made to incorporate properties of the occlusal contact, whether from opposing dentition or intervening food particles. All these features are demonstrated on critical-load master diagrams. A characteristic feature of the damage evolution is the gradual evolution of each mode with increasing load, so that failure is generally a prolonged rather than abrupt event. This accounts for the remarkable damage tolerance of natural teeth. The equations may enable basic predictions of tooth responses for humans and animals under a variety of specified dietary and functional conditions.     

Surface structure study of biological calcium phosphate apatite crystals from human tooth enamel

A surface-structure study of human tooth enamel crystals has been carried out by high-resolution electron microscopy (HREM). The surfaces of several crystals have been examined and the surfaces of a single crystal are described here. The observations made on this crystal are similar to the observations made on the other crystals, although the difference of morphology in the crystals observed indicates that the growth control by matrix proteins takes only place on the [011 macro 0] surfaces of the crystals. The crystal described here is oriented along the [112 macro 0] direction, and the following surfaces have been analyzed: (11 macro 00), (011 macro 1), (01 macro 11) and (0001). A subsurface reconstruction is observed just below the surface of the crystal not bonded to the matrix and lying above the supporting film. Observation of the matrix surrounding the crystal shows the existence of poorly crystalline phases with a structure close to that of hydroxyapatite (OHAP). Finally, a comparison of the images of the (011 macro 0) surface with computer-simulated images calculated for several models of the OHAP surface structure shows that the surface itself is stoichiometric and contains both calcium and phosphate groups. The knowledge of the binding sites of proteins on biomineral crystals such as the ones found in human tooth enamel is of prime importance for the understanding of their growth process. In this study, the first structure determination of the surface of human tooth enamel crystals is presented.