Development of the primary and secondary jaw joints in the mouse.

This study assesses the morphogenesis of the primary and secondary jaw joints. A collection of 11 murine heads, ranging from prenatal stage E13.5 to postnatal stage P10, were prepared as histological serial sections (thickness 8-10 µm) and stained conventionally in order to examine them with light microscopy. Next, the regions of the developing temporomandibular joint and the middle ear ossicles were three dimensionally reconstructed using AnalySIS® software. This study gained new insight into the spatio-temporal development of the temporomandibular joint and the auditory ossicles. Furthermore, we newly visualized in 3D that during the developmental period from stages E16 to P4 two morphologically well-functional joints (the primary and secondary jaw joints) exist on either side and are mechanically connected via Meckel's cartilage. Potential separation mechanisms of these two joints are discussed and options for mathematical analysis are suggested.

Numerical investigations of bone remodelling around the mouse mandibular molar primordia.

The formation of the alveolar bone, which houses the dental primordia, and later the roots of tooth, may serve as a model to approach general questions of alveolar bone formation. In this respect, this study aimed to investigate the potential interactions between the alveolar bone formation and tooth eruption by using finite element (FE) methods, and to figure out whether the expanding tooth systems induce shear stresses that lead to alveolar bone formation. 3D geometric surface models were generated from the 3D histological data of the heads of mice (C57 Bl/6J) ranging from stages embryonic (E) to postnatal (P) stages E15 to P20 using the reconstruction software 3-Matic. Bone, dentin, enamel and dental follicle around the primordia were generated and converted into 3D FE models. Models were imported into the FE software package MSC.Marc/Mentat. As material parameters of embryonic dentine, pulp, enamel, dental follicle, and bony structures basically are unknown, these were varied from 1% to 100% of the corresponding known material parameters for humans and a sensitivity analysis was performed. Surface loads were applied to the outside surface of dental follicle ranging from 0.1 to 5.0N/mm2. The validity of the model was analysed by comparing the activity pattern of the alveolar bone as determined in the histological study with the loading pattern from the numerical analysis. The results show that when varying the surface loads, the distribution of shear stresses remained same, and while varying the material properties of the hard tissues, the location of highest shear stresses remained stable. Comparison of the histologically determined growth regions with the distribution of shear stresses computed in the numerical model showed a very close agreement. The results provide a strong proof to support Blechschmidt's hypothesis that the bone in general is created under the influence of shear forces.

Structural, mechanical and chemical evaluation of molar-incisor hypomineralization-affected enamel: A systematic review.

OBJECTIVES: To systematically assess and contrast reported differences in microstructure, mineral density, mechanical and chemical properties between molar-incisor-hypomineralization-affected (MIH) enamel and unaffected enamel. METHODS: Studies on extracted human teeth, clinically diagnosed with MIH, reporting on the microstructure, mechanical properties or the chemical composition and comparing them to unaffected enamel were reviewed. Electronic databases (PubMed, Embase and Google Scholar) were screened; hand searches and cross-referencing were also performed. RESULTS: Twenty-two studies were included. Fifteen studies on a total of 201 teeth investigated the structural properties, including ten (141 teeth) on microstructure and seven (60 teeth) on mineral density; six (29 teeth) investigated the mechanical properties and eleven (87 teeth) investigated the chemical properties of MIH-affected enamel and compared them to unaffected enamel. Studies unambiguously found a reduction in mineral quantity and quality (reduced Ca and P content), reduction of hardness and modulus of elasticity (also in the clinically sound-appearing enamel bordering the MIH-lesion), an increase in porosity, carbon/carbonate concentrations and protein content compared to unaffected enamel. FINDINGS: were ambiguous with regard to the extent of the lesion through the enamel to the enamel-dentin junction, the Ca/P ratio and the association between clinical appearance and defect severity. CONCLUSIONS: There is an understanding of the changes related to MIH-affected enamel. The association of these changes with the clinical appearance and resulting implications for clinical management are unclear. CLINICAL SIGNIFICANCE: MIH-affected enamel is greatly different from unaffected enamel. This has implications for management strategies. The possibility of correlating the clinical appearance of MIH-affected enamel with the severity of enamel changes and deducing clinical concepts (risk stratification etc.) is limited.

Chondral ossification centers next to dental primordia in the human mandible: A study of the prenatal development ranging between 68 to 270mm CRL.

The human mandible is said to arise from desmal ossification, which, however, is not true for the entire body of the mandible: Meckel's cartilage itself is prone to ossification, at least its anterior part in the canine and incisor region. Also, within the coronoid and in the condylar processes there are cartilaginous cores, which eventually undergo ossification. Furthermore, there are a number of additional single cartilaginous islets arising in fetuses of 95mm CRL and more. They are located predominantly within the bone at the buccal sides of the brims of the dental compartments, mostly in the gussets between the dental primordia. They become wedge-shaped or elongated with a diameter of around 150-500µm and were also found in older stages up to 225mm CRL, which was the oldest specimen used in this study. This report is intended to visualize these single cartilaginous islets histologically and in 3-D reconstructions in stereoscopic images. Although some singular cartilaginous tissue within the mandible may be remains of the decaying Meckel's cartilage, our 3-D reconstructions clearly show that the aforementioned cartilaginous islets are independent thereof, as can be derived from their separate locations within the mandibular bone. The reasons that lead to these cartilaginous formations have remained unknown so far.

The remodeling pattern of human mandibular alveolar bone during prenatal formation from 19 to 270mm CRL.

The underlying mechanisms of human bone morphogenesis leading to a topologically specific shape remain unknown, despite increasing knowledge of the basic molecular aspects of bone formation and its regulation. The formation of the alveolar bone, which houses the dental primordia, and later the dental roots, may serve as a model to approach general questions of bone formation. Twenty-five heads of human embryos and fetuses (Radlanski-Collection, Berlin) ranging from 19mm to 270mm (crown-rump-length) CRL were prepared as histological serial sections. For each stage, virtual 3D-reconstructions were made in order to study the morphogenesis of the mandibular molar primordia with their surrounding bone. Special focus was given to recording the bone-remodeling pattern, as diagnosed from the histological sections. In early stages (19-31mm CRL) developing bone was characterized by appositional only. At 41, in the canine region, mm CRL bony extensions were found forming on the bottom of the trough. Besides general apposition, regions with resting surfaces were also found. At a fetal size of 53mm CRL, septa have developed and led to a compartment for canine development. Furthermore, one shared compartment for the incisor primordia and another shared compartment for the molars also developed. Moreover, the inner surfaces of the dental crypts showed resorption of bone. From this stage on, a general pattern became established such that the compartmentalizing ridges and septa between all of the dental primordia and the brims of the crypts were noted, and were due to appositional growth of bone, while the crypts enlarged on their inner surfaces by resorption. By 160mm CRL, the dental primordia were larger, and all of the bony septa had become reduced in size. The primordia for the permanent teeth became visible at 225mm CRL and shared the crypts of their corresponding deciduous primordia.

Osteogenic Profile of Mesenchymal Cell Populations Contributing to Alveolar Bone Formation.

Teeth develop within the surrounding periodontal tissues, involving the alveolar bone, periodontal ligament and cementum. The alveolar bone originates through the process of intramembranous ossification involving mesenchymal cells from the tooth germ. As most available data are related to endochondral ossification, we examined the molecular background of alveolar bone development. We investigated the osteogenic profile of mesenchymal cells dissected from mouse mandible slices at the stage of early alveolar bone formation. Relative monitoring of gene expression was undertaken using PCR Arrays; this included the profiles of 84 genes associated with osteogenesis. To examine the tooth-bone interface, stages with detectable changes in bone remodelling during development (E13.0, E14.0 and E15.0) were chosen and compared with each other. These results showed a statistically significant increase in the expression of the genes Fgf3, Ctsk, Icam-1, Mmp9, Itga3 and Tuft1, and of a wide range of collagens (Col1a2, Col3a1, Col7a1, Col12a1, Col14a1). Decreased expression was detected in the case of Col2a1, Sox9, Smad2 and Vegfb. To confirm these changes in gene expression, immunofluorescence analyses of Mmp9 and Sox9 proteins were performed in situ. Our research has identified several candidate genes that may be crucial for the initiation of alveolar bone formation and is the basis for further functional studies.

Morphogenesis of the compartmentalizing bone around the molar primordia in the mouse mandible during dental developmental stages between lamina, bell-stage, and root formation (E13-P20).

Despite increasing knowledge of the basic molecular aspects of bone formation and its regulation, the mechanisms of bone morphogenesis leading to a topologically specific shape remain unknown. The formation of the alveolar bone, which houses the dental primordia and later, the dental roots, may serve as a model to understand the formation of bone form in general. Thirty-eight heads of mice (C57 Bl/6J) ranging from stages E13-P20 were used to prepare histological serial sections. For each stage, virtual 3D-reconstructions were made in order to study the morphogenesis of the mandibular molar primordia concomitantly with their surrounding bone. Special focus was given to recording the remodeling pattern. It has been shown that, in early stages (E13, E14), bone formation is characterized by apposition only. In stage E15, the bony crypt around the dental primordia is remodeled mostly by resorption of bone. In stage E18, the bone remodeling pattern shows resorption all along the bony gutter, which houses the molar primordia. The medial and lateral margins are characterized by apposition. At birth (stage P0), a bony septum has begun to form between the primordium m1 and of m2, arising from both sides and characterized by pure apposition of bone. In stage P4, the crypts of m1 and m2, and also that of m3, show bone resorption inside, while the medial and lateral bony margins show apposition of bone throughout. Generally, during development, the bone gradually encapsulates the dental primordia, in such a way that the bone reaches over the dental primordia and leaves only a continuous longish opening of about 200µm width. The opening at the occlusal surface of m1, at the time of eruption, starting at stage P14, appears to have increased in size again. The distance between bone and dental primordium undergoes change during development. In erupted molars, it is around 100µm, during early developmental stages, it may be as less as 20µm. These data show the inevitability of bone remodeling.

Effect of methotrexate upon antigen-induced arthritis of the rabbit temporomandibular joint.

BACKGROUND: Juvenile idiopathic arthritis (JIA) of the temporomandibular joint (TMJ) can cause severe growth disturbances of the craniomandibular system. Antigen-induced arthritis (AIA) of the rabbit TMJ is simulating the inflammatory process of the TMJ in JIA. The aim of this study was to investigate the effect of a systemic administration of methotrexate (MTX) on AIA in rabbits by means of three different histological staining methods. METHODS: After sensitization, a bilateral arthritis of the TMJ was induced by an intra-articular administration of ovalbumin in 12 New Zealand white rabbits aged 10 weeks. From the 13th week of age, six of the 12 rabbits received weekly intramuscular injections of MTX, and the other six animals remained without therapy. Another six animals served as controls, receiving no treatment or intra-articular injections at all. After euthanasia at the age of 22 weeks, all TMJs were retrieved en bloc. Sagittal sections were cut and stained with haematoxylin-eosin (H-E), Safranin-O for the evaluation of the Mankin score and tartrate-resistant acid phosphatase (TRAP). RESULTS: In the arthritis group, a chronic inflammation with degeneration of the articular cartilage was visible. In the MTX group, the signs of cartilage degeneration were significantly reduced compared with the arthritis group. In contrast, the joints in the control group were inconspicuous. A correlation between the Mankin score and TRAP-positive cells could be found. CONCLUSIONS: Systemic administration of MTX seems to have a positive effect upon the inflammatory process in the rabbit TMJ but fails to eliminate the sign of arthritis completely.

Distribution of BMP6 in the alveolar bone during mouse mandibular molar eruption.

Eruption requires synchrony of the tooth with the surrounding tissues, particularly the bone. One important step during eruption is remodelling of the alveolar bone at the base of the tooth and along the roots. Expression of BMP6 was reported to be increased in the basal half of the dental follicle prior to eruption and inhibition of BMP6 affected bone formation at the base of the alveolar crypt. The aim of this study was to further investigate BMP6 protein in relation to tooth eruption and the corresponding bone remodelling using temporospatial correlations of BMP6 localization with morphogenetic events (proliferation, differentiation, apoptosis and bone apposition/resorption), other BMPs (BMP2 and BMP7) and three-dimensional images of tooth-bone development. BMP6 expression pattern was mapped in the mandibular molar teeth and related structures around eruption. Localization of BMP6 dominated in osteoblasts, in regions of bone formation within the alveolar crypt. These findings positively correlated with proliferation at the tooth base region, osteocalcin expression in the osteoblasts/osteocytes and BMP2 and BMP7 presence in the alveolar bone surrounding the tooth. Osteoclast activity and apoptotic elimination in the root region gradually decreased before eruption and totally ceased at eruption stages. Generally, BMP6 positively correlated with BMP2, BMP7 and osteocalcin-positive osteoblasts, and areas of bone remodelling. Moreover, BMP6 was found in the periodontium and cementoblasts. BMP6 expression in the alveolar bone accompanied tooth eruption. Notably, the expression pattern of BMP6 in the bone did not differ around individual molar teeth at the same stage of development. The expression of BMP6 in periodontal ligaments may contribute to interaction between the tooth and bone during the eruption and anchoring process.

Fluorosed mouse ameloblasts have increased SATB1 retention and Gαq activity.

Dental fluorosis is characterized by subsurface hypomineralization and increased porosity of enamel, associated with a delay in the removal of enamel matrix proteins. To investigate the effects of fluoride on ameloblasts, A/J mice were given 50 ppm sodium fluoride in drinking water for four weeks, resulting serum fluoride levels of 4.5 µM, a four-fold increase over control mice with no fluoride added to drinking water. MicroCT analyses showed delayed and incomplete mineralization of fluorosed incisor enamel as compared to control enamel. A microarray analysis of secretory and maturation stage ameloblasts microdissected from control and fluorosed mouse incisors showed that genes clustered with Mmp20 appeared to be less downregulated in maturation stage ameloblasts of fluorosed incisors as compared to control maturation ameloblasts. One of these Mmp20 co-regulated genes was the global chromatin organizer, special AT-rich sequence-binding protein-1 (SATB1). Immunohistochemical analysis showed increased SATB1 protein present in fluorosed ameloblasts compared to controls. In vitro, exposure of human ameloblast-lineage cells to micromolar levels of both NaF and AlF3 led to a significantly increase in SATB1 protein content, but not levels of Satb1 mRNA, suggesting a fluoride-induced mechanism protecting SABT1 from degradation. Consistent with this possibility, we used immunohistochemistry and Western blot to show that fluoride exposed ameloblasts had increased phosphorylated PKCα both in vivo and in vitro. This kinase is known to phosphorylate SATB1, and phosphorylation is known to protect SATB1 from degradation by caspase-6. In addition, production of cellular diacylglycerol (DAG) was significantly increased in fluorosed ameloblasts, suggesting that the increased phosphorylation of SATB1 may be related to an effect of fluoride to enhance Gαq activity of secretory ameloblasts.

Sexual dimorphism in teeth? Clinical relevance.

Many morphometric studies show a sexual dimorphism in human teeth. We wanted to know whether it is possible to determine the sex of an individual if only the anterior teeth are visible. Fifty intraoral photographs showing the front tooth region of female and male individuals (age: from 7 to 75 years) were randomly arranged in actual size on a questionnaire. The lip region was covered in each case. Besides "female" and "male", one was also able to check "?" if undecided. The questionnaires were distributed to 50 expert test persons (dentists, dental technicians, dental assistants, and students of dental medicine) and to 50 laymen and were all returned for evaluation. Although the correct sex was recognized on single photographs to a maximum of 76%, it was incorrect in 69% on other photographs. Altogether, the statistical evaluation showed that in most cases, the sex was only recognized correctly by one half, and incorrect by the other half. It can be concluded that a sexual dimorphism of human teeth-although measurable morphometrically-could not be recognized visually on the basis of photographs of the front tooth region. Neither experts in the field of dentistry nor laymen were able to properly distinguish between male and female teeth.

Morphogenesis and bone integration of the mouse mandibular third molar.

The mouse third molar (M3) develops postnatally and is thus a unique model for studying the integration of a non-mineralized tooth with mineralized bone. This study assessed the morphogenesis of the mouse M3, related to the alveolar bone, comparing M3 development with that of the first molar (M1), the most common model in odontogenesis. The mandibular M3 was evaluated from initiation to eruption by morphology and by assessing patterns of proliferation, apoptosis, osteoclast distribution, and gene expression. Three-dimensional reconstruction and explant cultures were also used. Initiation of M3 occurred perinatally, as an extension of the second molar (M2) which grew into a region of soft mesenchymal tissue above the M2, still far away from the alveolar bone. The bone-free M3 bud gradually became encapsulated by bone at the cap stage at postnatal day 3. Osteoclasts were first visible at postnatal day 4 when the M3 came into close contact with the bone. The number of osteoclasts increased from postnatal day 8 to postnatal day 12 to form a space for the growing tooth. The M3 had erupted by postnatal day 26. The M3, although smaller than the M1, passed through the same developmental stages over a similar time span but showed differences in initiation and in the timing of bone encapsulation.

Tooth-bone morphogenesis during postnatal stages of mouse first molar development.

The first mouse molar (M1) is the most common model for odontogenesis, with research particularly focused on prenatal development. However, the functional dentition forms postnatally, when the histogenesis and morphogenesis of the tooth is completed, the roots form and the tooth physically anchors into the jaw. In this work, M1 was studied from birth to eruption, assessing morphogenesis, proliferation and apoptosis, and correlating these with remodeling of the surrounding bony tissue. The M1 completed crown formation between postnatal (P) days 0-2, and the development of the tooth root was initiated at P4. From P2 until P12, cell proliferation in the dental epithelium reduced and shifted downward to the apical region of the forming root. In contrast, proliferation was maintained or increased in the mesenchymal cells of the dental follicle. At later stages, before tooth eruption (P20), cell proliferation suddenly ceased. This withdrawal from the cell cycle correlated with tooth mineralization and mesenchymal differentiation. Apoptosis was observed during all stages of M1 postnatal morphogenesis, playing a role in the removal of cells such as osteoblasts in the mandibular region and working together with osteoclasts to remodel the bone around the developing tooth. At more advanced developmental stages, apoptotic cells and bodies accumulated in the cell layers above the tooth cusps, in the path of eruption. Three-dimensional reconstruction of the developing postnatal tooth and bone indicates that the alveolar crypts form by resorption underneath the primordia, whereas the ridges form by active bone growth between the teeth and roots to form a functional complex.

Interdental spacing and orthodontic treatment in competitive athletes: clues to doping with growth hormones?

UNLABELLED: The aim of this report is to examine clues of a suspected link between the artificial ingestion of human growth hormone (rh- GH) and resulting interdental spaces in adult athletes. We conducted an electronic search in the German-language versions of the search engines Google and Google Scholar as well as in the database PubMed. While no explicit articles could be identified in PubMed, the search in Google and Google Scholar produced 1370 and 6 hits, respectively. Original quotes from 20 sources show that in the media the wearing of orthodontic multibracket appliances among athletes is largely attributed to changes in tooth position as a consequence of the illegal ingestion of rhGH. On the other hand, there are few references to the possibility that orthodontic treatments with fixed appliances might be carried out for reasons unrelated to doping. CONCLUSION: A definitive assessment of this issue is not possible at present. In view of its major importance of the subject, the relationships depicted here should be investigated in greater depth.

An atlas of prenatal development of the human orofacial region.

There are several atlases available showing prenatal human development. However, none is focused on prenatal orofacial development during maxillary and mandibular bone formation. These events, together with dental development and formation of the temporomandibular joint, take place during several fetal stages. While photographic atlases are limited to depicting the outer shape, and atlases based on histological sections only show a series of single sections, an atlas based on three-dimensional reconstructions from serial sections can show both the outer skin and the structures underneath, which can be electronically dissected layer by layer. In this Focus article, we present our atlas on prenatal human orofacial development, which is accessible online at the Journal's website.

Ameloblast differentiation in the human developing tooth: effects of extracellular matrices.

Tooth enamel is formed by epithelially-derived cells called ameloblasts, while the pulp dentin complex is formed by the dental mesenchyme. These tissues differentiate with reciprocal signaling interactions to form a mature tooth. In this study we have characterized ameloblast differentiation in human developing incisors, and have further investigated the role of extracellular matrix proteins on ameloblast differentiation. Histological and immunohistochemical analyses showed that in the human tooth, the basement membrane separating the early developing dental epithelium and mesenchyme was lost shortly before dentin deposition was initiated, prior to enamel matrix secretion. Presecretary ameloblasts elongated as they came into contact with the dentin matrix, and then shortened to become secretory ameloblasts. In situ hybridization showed that the presecretory stage of odontoblasts started to express type I collagen mRNA, and also briefly expressed amelogenin mRNA. This was followed by upregulation of amelogenin mRNA expression in secretory ameloblasts. In vitro, amelogenin expression was upregulated in ameloblast lineage cells cultured in Matrigel, and was further up-regulated when these cells/Matrigel were co-cultured with dental pulp cells. Co-culture also up-regulated type I collagen expression by the dental pulp cells. Type I collagen coated culture dishes promoted a more elongated ameloblast lineage cell morphology and enhanced cell adhesion via integrin alpha2beta1. Taken together, these results suggest that the basement membrane proteins and signals from underlying mesenchymal cells coordinate to initiate differentiation of preameloblasts and regulate type I collagen expression by odontoblasts. Type I collagen in the dentin matrix then anchors the presecretary ameloblasts as they further differentiate to secretory cells. These studies show the critical roles of the extracellular matrix proteins in ameloblast differentiation.

[Genes, forces and forms: mechanical aspects of prenatal craniofacial development]. / Gènes, forces et formes: aspects mécaniques du développement cranio-facial prénatal.

Current knowledge of molecular signaling during craniofacial development is advancing rapidly. We know that cells can respond to mechanical stimuli by biochemical signaling. Thus, the link between mechanical stimuli and gene expression has become a new and important area of the morphological sciences. This field of research seems to be a revival of the old approach of developmental mechanics, which goes back to the embryologists His [36], Carey [13, 14], and Blechschmidt [5]. These researchers argued that forces play a fundamental role in tissue differentiation and morphogenesis. They understood morphogenesis as a closed system with living cells as the active part and biological, chemical, and physical laws as the rules. This review reports on linking mechanical aspects of developmental biology with the contemporary knowledge of tissue differentiation. We focus on the formation of cartilage (in relation to pressure), bone (in relation to shearing forces), and muscles (in relation to dilation forces). The cascade of molecules may be triggered by forces, which arise during physical cell and tissue interaction. Detailed morphological knowledge is mandatory to elucidate the exact location and timing of the regions where forces are exerted. Because this finding also holds true for the exact timing and location of signals, more 3D images of the developmental processes are required. Further research is also required to create methods for measuring forces within a tissue. The molecules whose presence and indispensability we are investigating appear to be mediators rather than creators of form.

New carbide finishing burs to reduce polishing efforts of light-cured restorations.

OBJECTIVE: To compare, under laboratory conditions, the efficacy of different polishing systems for various light-curing restorative materials. METHOD AND MATERIALS: Class 5 cavities were prepared in 65 human teeth and filled with 5 different light-curing restorative materials. To increase the number of samples, an extra 585 Class 5 cavities were made in resin disks and filled with the 5 different restorative materials. All restorations were polished with 13 polishing procedures commonly used in dental practice. The restoration surfaces were evaluated micromorphologically by means of SEM. A quantitative analysis of the polishing result was performed with the surface tester with waveline profilometry. The time required to achieve a polished surface was also measured. RESULTS: Surfaces of all 5 restorative materials showed a satisfactory finish after treatment. SEM analysis and profilometry showed good surface qualities with large smooth, homogenous areas with low roughness. All polished surfaces were smoother than natural dental enamel. Sof-Lex, Super Snap, and Bush composite finished set 5430 systems showed the best areas with a smooth and homogenous surface. The extra effort required with several systems resulted in a smooth final restoration. CONCLUSION: The surface produced by the carbide finisher H134Q was rough, so it is useful for effective contouring of restorations. Application of the finishers H282K and H22ALGK led to a high share in good surface quality, which could be enhanced by using polishing paste as a final working step. The instruments H22GK and the systems One Gloss and Enhance were not recommendable. In comparison, the combination of H134Q and H22ALGK was effective, at the same time preserving tissue and saving time in finishing light-cured Class 5 restorations. The use of polishing paste as a final step is recommended.

Insular dentin formation pattern in human odontogenesis in relation to the scalloped dentino-enamel junction.

This study is a first report on the modality of early dentin formation in respect to the scalloped pattern of the dentino-enamel junction (DEJ). We applied scanning electron microscopy (SEM), transmission electron microscopy (TEM), histological serial sections, and three-dimensional (3D) reconstructions. TEM and SEM showed scallops and secondary scallops on the DEJ of deciduous dental primordia and on deciduous teeth with the enamel cap removed. This peculiar outline of the DEJ requires a specific dentin formation pattern; histological sections showed that dentin formation began at the brims of the scallops, seen as triangular spikes in serial sections. The dentin formation front was not uniform; instead, it was characterized by multiple, insular forming centers, as revealed by our 3D reconstructions. As thicker dentin layers formed, the islands became confluent. Factors are discussed, which may lead to crimpling of the inner enamel epithelium, and maintained as the scalloped pattern of the DEJ develops. Signaling patterns in accordance with the insular dentin formation are unknown so far.

Developmental movements of the inner enamel epithelium as derived from micromorphological features.

It was the purpose of this article to analyze the (micro) morphological structure of enamel at different stages of development in order to deduce movement patterns of ameloblasts during formation of the human dental primordium. Developing enamel and overlying ameloblasts were dried and fractured for scanning electron microscopy (SEM) and sectioned for transmission electron microscopy (TEM). Specimens of human permanent enamel were either fractured and/or ground and etched to visualize the enamel rods. All specimens were viewed by SEM. Moreover, three-dimensional reconstructions were made from serial ground sections of enamel blocks to follow the enamel rods for a longer distance. In addition, the outline of the dentino-enamel junction was analyzed under the SEM after removal (using nitric acid) of the enamel cap, and in serial histological sections. Two basic movements of the inner enamel epithelium can be derived from the micromorphological features: (i) the scalloped dentino-enamel junction may be a consequence of a bulged inner enamel epithelium owing to initial spatial impediment; and (ii) the undulating path of the enamel rods may be a consequence of unequal growth of the cells in the cervical loop.