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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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Structured Abstract Author – Radlanski RJ Objectives – The formation of the dental primordial should be visualized with special reference to characteristic differences for each single primordium. Until today, it has not been clear how traffic of the ameloblasts is controlled, how the folding pattern of the occlusal relief is generated or how the enamel production is terminated at the enamel surface. Design – Using computer-aided reconstructions from histological serial sections, the dental primordial were visualized and, using fractured enamel specimens, the traces of each single ameloblast were followed by means of scanning electron microscopy. In this way, the developmental movements of the inner enamel epithelium can be reconstructed. Results – Gathering morphological knowledge, three-dimensional polygon sets of shape data were input into a computer workstation and animated by means of the software Soft Image (Microsoft). Conclusions – The development of the human primary and permanent dentition was animated to simulate growth. [ABSTRACT FROM AUTHOR]

Structured Abstract Author – Radlanski RJ Objectives – Basic research concerning craniofacial development presently runs along two pathways, namely the molecular and the morphometric. This gap needs to be bridged. Design – Using histological serial sections of human fetuses computer-aided three-dimensional reconstructions were made (Software Analysis, SIS) with special focus given to all anatomical structures of the orofacial region of the growing head. Results – All reconstructions can be viewed from any rotation and they are available for virtual dissection according to anatomical rules. As an example, the prenatal development of the human mandible with the formation of the mental foramen therein is described. Furthermore, the spatial arrangement of bone, cartilage and nerves is presented in three dimensions in different developmental stages. The interaction of tissues with possible morphogenetic interaction is discussed. Conclusions – This work serves as a reference system for prenatal development in comparison with pathological development. [ABSTRACT FROM AUTHOR]

Background: During physiological function of the temporomandibular joint, we have to rely only on elastic structures (in particular the bilaminar zone) for repositioning of the articular disc. A real muscle, however, would be functionally more reasonable. In patients, a decrease of this elasticity is possibly one of the reasons for temporomandibular joint malfunctions, which affect between 16% and 36% of the population., Method: This study assesses the morphogenesis of the murine (Mus musculus) temporomandibular joint with particular regard to the masticatory muscles, to throw light on this topic. To that end, a collection of 11 murine heads ranging from prenatal stages E13.25 to E20 was used and early postnatal stages P0 to P4, which were prepared as histological sections (thickness 8-10 µm) and stained conventionally in order to examine them with light microscopy. Next, the temporomandibular joint and selected surrounding structures, along with the masticatory muscles, were threedimensionally reconstructed using analySIS® software. Subsequently, specific morphometric analyses were performed., Results and Conclusions: The evaluation of the results led to the following conclusions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier GmbH.)

Craniofacial morphogenesis is an intricate developmental process in 3D, which therefore merits visualization and investigation in 3D. To better understand the process, we utilize μCT imaging, and describe a method to calibrate each cone beam μCT individually. Calibration is necessary, because during development, fetuses undergo tissue differentiation, which affects the acquisition process for radiographic images. Additionally, tissue fixation and conservation agents may influence the physical properties of the specimens and may affect image acquisition. After taking a μCT scan from each specimen, we separated a horizontal slice from each neck (which is inconsequential to our question with relation to the whole head). These neck specimens were prepared as horizontal histological serial sections and stained. With these as a reference, the μCT visualization parameters could be adjusted until they matched the selected virtual section planes, which correspond exactly to the planes of the histological sections with a precision (pixel size) of 0.69μm., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Radlanski RJ, Renz H. An atlas of prenatal development of the human orofacial region. Eur J Oral Sci 2010; 118: 321–324. © 2010 The Authors. Journal compilation © 2010 Eur J Oral Sci 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. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to investigate the thickness of acellular extrinsic fibre cementum (AEFC) at four root positions of anterior and posterior teeth with special focus on functional aspects. Furthermore, the correlations between cementum thickness and chronological age and sex are investigated., Background: While numerous studies confirm continuous cementum apposition with age, masticatory forces as well as physiological and orthodontically induced tooth movements also have the potential to affect tooth cementum thickness., Materials and Methods: Undecalcified teeth were embedded in resin and transverse-sectioned in the cervical third of the root. Two sections per root were selected, and digital images at four positions were obtained (mesial, distal, oral, and vestibular) using light microscopy. The AEFC thickness of 99 teeth (anterior = 66, posterior = 33, male = 54, female = 45) were measured in both sections. The differences in mean values between root positions and the association of root position variation with tooth type, age, sex, and subject as well as the overall effects of age and sex were analysed using a mixed model., Results: First incisors and canines showed the greatest mean AFEC thickness, in contrast to premolars which had the lowest values. Differences were found across the four root positions, with a pattern varying considerably between anterior and posterior teeth and between maxilla and mandible in the anterior teeth. An interaction between root position and subject pointed to the existence of an individual component in the variation of AEFC thickness across the four root positions. There was an age trend with an almost linear increase in cementum thickness of 1 μm per year. Overall, females tended to exhibit a significantly lesser AEFC thickness compared to males., Conclusions: Distinct differences in the pattern of thickness values across the four root positions in anterior and posterior teeth support the assumption that the AEFC is strongly affected by functional processes. In addition to sex-specific differences and age-related trends, the root position variation of AEFC thickness varies from individual to individual., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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., (Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.)

Background: Cleft lip and palate (CLP) represents a group of malformations of unknown etiology but similar phenotypes. This implies consequences for the diagnostics, therapy, prevention, prognosis and risk estimation., Objective: Definition of CLP subtypes and the embryonic development, clarification of correlations and differences between entities using epidemiological data, overview of the present state of genetic analyses, correlation to syndromes, sequences and associations and resulting consequences for clinical practice., Material and Methods: Update on embryological development of the face, summary of epidemiological and genetic studies and considerations on pedopathological and forensic aspects., Results: Syndromic and non-syndromic CLP exhibit different and highly variable etiologies, therapeutic needs and prognosis. A thorough understanding is mandatory to distinguish between the different subgroups. In addition to specific aspects of CLP for the pediatric (forensic) pathologist this article provides an overall view of the topic which aims to help understand these malformations.

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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., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

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., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Radlanski RJ, Renz H. Developmental movements of the inner enamel epithelium as derived from micromorphological features. Eur J Oral Sci 2006; 114 (Suppl. 1): 343–348 © Eur J Oral Sci, 2006 --> 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. [ABSTRACT FROM AUTHOR]

Dental hard tissues are formed particularly by odontoblasts (dentin) and ameloblasts (enamel). Whereas the reparation of dentin is often observed, enamel does not regenerate in most species. However, in mouse incisor, a population of somatic stem cells in the cervical loop is responsible for the incisor regeneration. Understanding of the specificities of these cells is therefore of an interest in basic research as well as regenerative therapies. The Myb transcription factors are involved in essential cellular processes. B-Myb is often linked to the stem cell phenotype, and c-Myb expression marks undifferentiated and proliferating cells such as the stem cells. In the presented study, temporo-spatial expression of B-Myb and c-Myb proteins was correlated with localisation of putative somatic stem cells in the mouse incisor cervical loop by immunohistochemistry. B-Myb expression was localised mostly in the zone of transit-amplifying cells, and c-Myb was found in the inner enamel epithelium, the surrounding mesenchyme and in differentiated cells. Taken together, neither B-Myb nor c-Myb was exclusively present or abundant in the area of the incisor stem cell niche. Their distribution, however, supports recently reported novel functions of c-Myb in differentiation of hard tissue cells., (© 2014 Blackwell Verlag GmbH.)

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., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

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., (© 2015 S. Karger AG, Basel.)

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 the tumor necrosis factor-alpha (TNF-α) antagonist etanercept on AIA in rabbits by means of three different histological staining methods., Methods: After sensitization, a bilateral arthritis of the TMJ was induced and maintained by repeated intra-articular administrations of ovalbumin in 12 New Zealand white rabbits aged 10 weeks. From the 13th week of age, 6 of the 12 rabbits received weekly subcutaneous injections of etanercept, and the other 6 animals remained without therapy. Another 6 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 hematoxylin-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 etanercept group, the signs of cartilage degeneration were significantly reduced but present. In contrast, the joints in the control group were inconspicuous. A strong correlation between the Mankin score and TRAP-positive cells could be found., Conclusions: Antigen-induced arthritis causes severe damage in the TMJ of young rabbits. An improvement seems to be achievable by a systemic administration of etanercept., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Molar–incisor hypomineralization (MIH) is a qualitative enamel defect that is highly prevalent in children. It has been reported that patients with MIH have higher caries occurrence with an increased need and frequency of dental treatment compared to patients without MIH. The objective of this study was to analyze the association between MIH and a series of factors related to maternal health status during pregnancy and children´s medical history in early childhood. Methods: A retrospective study of cases (patients with MIH) and controls (patients without MIH) was designed between 2023 and 2024. A total of 280 children (cases = 140; controls = 140) aged 6 to 14 years (138 boys and 142 girls) were examined according to the European Academy of Pediatric Dentistry (EAPD) criteria for MIH. A survey was carried out with mothers regarding the potential exposure of their children to etiological factors of MIH. Possible prenatal and postnatal etiological factors were obtained through a personal interview with the patients' mothers. The statistical analysis was carried out with the contrast test and the chi-square test. Results: During pregnancy, folic acid consumption, alcohol intake, systemic viral and/or bacterial infections, and gestational diabetes were statistically significantly related to MIH, as were breastfeeding, asthma, and corticosteroid consumption during childhood. Conclusions: Although there are different factors that may have statistically significant relationships with MIH, they cannot be predicted. Therefore, longitudinal studies, with a large sample size, are needed to determine the influence of prenatal and postnatal factors on the prevalence and severity of MIH in children. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the association between the frequency and severity of dental fluorosis and Molar Incisor Hypomineralization (MIH) in 8-12-year-old schoolchildren living in an area with a high concentration of fluoride in the drinking water. Methods: The present cross‑sectional study was conducted on Mexican children (n = 573) selected from one community presenting a drinking water fluoride concentration of 1.39 ppm/F. The prevalence of dental fluorosis was ascertained using the Thylstrup and Fejerskov Index (TFI). The presence and severity of MIH was evaluated using the European Academy of Pediatric Dentistry (EAPD) criteria. A multinomial regression model was used to estimate the odds ratio (OR) and the 95% confidence intervals (CI), using the severity of MIH as the result. Results: The prevalence of MIH was 37.7% and, by severity, was 16.1% mild, 14.3% moderate, and 7.3% severe. The prevalence of dental fluorosis in permanent dentition was 70.9% (TFI ≥1) and, by severity, was 29.2% (TFI = 0), 45.6% (TFI 1–3) and 25.3% (TFI ≥4), while 54.5% of subjects were found to have poor oral hygiene. Schoolchildren with fluorosis (TFI ≥4) were 49% less likely [OR = 0.51; p = 0.025] to present mild MIH than children with fluorosis (TFI <4). Similarly, children with fluorosis (TFI ≥4) were 53% [OR = 0.47; p = 0.019] and 62% [OR = 0.38; p = 0.036] less likely to present moderate and severe MIH than children with fluorosis (TFI <4). Conclusion: An inverse relationship between the presence of fluorosis and MIH was found. The results obtained by the present study may contribute to both the early identification of disorders affecting the enamel and the creation and implementation of long-term oral health prevention, promotion, and intervention programs in the affected population. [ABSTRACT FROM AUTHOR]

Background: The molecular mechanisms and signaling pathways involved in tooth morphogenesis have been the research focus in the fields of tooth and bone development. However, the cell population in molars at the late bell stage and the mechanisms of hard tissue formation and mineralization remain limited knowledge. Results: Here, we used the rat mandibular first and second molars as models to perform single-cell RNA sequencing (scRNA-seq) analysis to investigate cell identity and driver genes related to dental mesenchymal cell differentiation during the late bell hard tissue formation stage. We identified seven main cell types and investigated the heterogeneity of mesenchymal cells. Subsequently, we identified novel cell marker genes, including Pclo in dental follicle cells, Wnt10a in pre-odontoblasts, Fst and Igfbp2 in periodontal ligament cells, and validated the expression of Igfbp3 in the apical pulp. The dynamic model revealed three differentiation trajectories within mesenchymal cells, originating from two types of dental follicle cells and apical pulp cells. Apical pulp cell differentiation is associated with the genes Ptn and Satb2, while dental follicle cell differentiation is associated with the genes Tnc, Vim, Slc26a7, and Fgfr1. Cluster-specific regulons were analyzed by pySCENIC. In addition, the odontogenic function of driver gene TNC was verified in the odontoblastic differentiation of human dental pulp stem cells. The expression of osteoclast differentiation factors was found to be increased in macrophages of the mandibular first molar. Conclusions: Our results revealed the cell heterogeneity of molars in the late bell stage and identified driver genes associated with dental mesenchymal cell differentiation. These findings provide potential targets for diagnosing dental hard tissue diseases and tooth regeneration. [ABSTRACT FROM AUTHOR]

Purpose Adhesive remnants removal is the last key step influencing orthodontic treatment outcomes. Four different clearance methods (CM) of orthodontic adhesive were evaluated to determine, which achieved the smoothest enamel surface in the shortest time. Materials and Methods 75 intact premolars extracted for orthodontic purposes were included, sixty had an orthodontic bracket bonded and subsequently removed, and fifteen served as the control group. Four CMs were used to clear the tooth surface of 15 premolars each: carbide bur (CB), carbide bur with titanium nitride surface treatment + fine carbide bur (CBCB), glass fiber-reinforced composite instrument (GFCB), zirconia bur + glass fiber-reinforced composite bur (ZBCB). The processing time was recorded. In ten premolars from each group, the enamel surface was evaluated by atomic force microscopy estimating mean roughness (Ra), roughness profile value (Rq), and roughness depth (Rt). Enamel Damage Index (EDI) was assessed with a scanning electron microscope on 5 remaining premolars. Results Significant differences were observed in all evaluated parameters - Ra (p<0.0001), Rq (p<0.0001), and Rt (p<0.0001). GFCB exhibited the smoothest surface in all parameters. The lowest EDI exhibited teeth treated by GFCB, however, the differences were not significant. Working with GFCB took the longest time (mean 116 s), and the shortest with CBCB (mean 49 s). Conclusion Using CB is the fastest clearance method, but the enamel surface roughness was highest. Clearing with a set of instruments CBCB proved to be a fast method with satisfying remaining enamel roughness. [ABSTRACT FROM AUTHOR]

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.

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.

Background: Past investigations of prenatal craniofacial growth have largely relied on histological sections. Few studies have taken measurements on three-dimensional representations (3D reconstruction, 3D CT, postmortem) or varying depth levels (ultrasound), and we know of no craniofacial growth studies done on cleared-and-stained specimens of whole fetal heads., Materials and Methods: This study comprised 14 human fetal head specimens cleared and stained with alizarin red and alcian blue. They had been stored in glycerol and represented weeks 8-12 of gestation, with crown-rump lengths ranging from 23-145 mm. These specimens were cephalometrically analyzed in norma frontalis and norma lateralis, which notably included the opportunity for side-to-side comparison., Results: As the cranial membrane bones progressively approached each other, the orbits, maxilla, and mandible gradually grew wider. Likewise, the sagittal dimensions of the maxilla and mandible increased continuously and synchronically. We noted side-to-side differences ranging from 2-5 mm. Another notable finding concerned the inclination of the maxilla relative to the cranial base, which increased more on the right than on the left side., Conclusion: This is the first investigation presenting side-to-side comparative measurements of human fetal head specimens. Such measurements are essential in the quest toward validating the findings of other imaging techniques such as CT or MRI and-most importantly-intrauterine sonography.

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.

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., (© 2011 Eur J Oral Sci.)

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., (© 2011 The Authors. Journal of Anatomy © 2011 Anatomical Society of Great Britain and Ireland.)

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.

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., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Not only are teeth essential for mastication, but also missing teeth are considered a social handicap due to speech and aesthetic problems, with a resulting high impact on emotional well-being. Several treatment procedures are currently available for tooth replacement with mostly inert prosthetic materials and implants. Natural tooth substitution based on copying the developmental process of tooth formation is particularly challenging and creates a rapidly developing area of molecular dentistry. In any approach, functional interactions among the tooth, the surrounding bone, and the periodontium must be established. Therefore, recent research in craniofacial genetics searches for mechanisms responsible for correct cell and tissue interactions, not only within a specific structure, but also in the context of supporting structures. A tooth crown that is not functionally anchored to roots and bone is useless. This review aims to summarize the developmental and tissue homeostatic aspects of the tooth-bone interface, from the initial patterning toward tooth eruption and lifelong interactions between the tooth and its surrounding alveolar bone.

Purpose: To compare the osseointegration process of the titanium dental implants with five different surface characteristics--sandblasted, sandblasted and acid-etched, hyaluronic acid--coated (HYA), hydroxyapatite-coated (HA), and machined--in an experimental sheep model at 1- and 3-month examinations. Materials and Methods: One hundred sixty dental implants were placed in the left and right tibias of 16 sheep. Five experimental groups were designed. Eight animals (80 implants) were used for biomechanical tests of reverse torque analysis and resonance frequency analysis (RFA). Eight of them (80 implants) were used for the evaluation of bone-to-implant contact (BIC) percentage in histomorphometric analysis. Forty of 80 implants (8 implants for each group) were used at 1-month examinations, and the remaining 40 (8 implants for each group) implants were used at 3-month examinations in the biomechanical test group and histomorphometric examination group, separately. Results: Intergroup analysis at the 3-month followup showed that the increase in the implant stability quotient (ISQ) value was statistically significant for only the HYA group (P < .05). According to ISQ values at 1 and 3 months, group HYA showed statistically higher values at the 1 and 3-month examinations (P < .05). Groups HYA and HA had statistically higher reverse torque values than other groups at the 1-month examination (P < .05). At the 3-month evaluation, the HYA group showed significantly higher reverse torque values compared to other groups (P < .05). The BIC values of the sandblasted and acid-etched, HYA, and HA groups were significantly higher than the sandblasted and machined groups at the 1- and 3-month examinations (P < .05). The BIC value for the HA group showed decreased value at the 3-month examination compared to the 1-month examination (P < .05). Conclusion: The RFA, reverse torque, and histomorphometric analysis at 1- and 3-month examinations show that dental implants coated with HYA may have increased potential for osseointegration compared to dental implants with sandblasted, sandblasted and acid-etched, machined, and HA-coated surfaces. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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.

The transformation of fibrinogen into fibrin is biologically activated in a complex multi-step process known as the coagulation cascade. This transformation can also be triggered by anodic surfaces. It has been suggested that this mechanism is a result of an electron transfer from the anode to the fibrinogen molecule resulting in the formation of fibrin. In this study we used this pathway to simultaneously deposit vital cells (fibroblasts and keratinocytes) and fibrin on micro structured gold electrodes. The electrodes were produced using a novel inverse inkjet-printing technology in combination with subsequent gold-sputtering, resulting in minimal structure-sizes of 35 microm (+/-6 microm). Cell deposition and fibrin-coagulation were found to occur on the anode only, following exactly the micro structured electrode surface. Successful deposition was limited by the minimal voltage (0.8 V) needed for the formation of fibrin and the maximum voltage (1.85 V) resulting in the deterioration of the Au-electrodes due to electrolysis and possible damaging of the deposited cells due to the formation of molecular chlorine. Furthermore, it was demonstrated that this technique is suitable to co-cultivate different cell types in a layered fashion. Subsequent to the electrically mediated anodic cell-protein deposition, cells were cultivated for up to 4 days and then characterized by vital fluorescence staining, methyl violet-staining and scanning electron microscopy. Cell-vitality was found to be dependent on the experimental setup; in this study non-vital cells were only observed, when sequentially depositing two different cell types. Finally, the coagulation mechanism was studied using HPLC, SDS-gel-chromatography and ATR/FTIR.

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.

As the use of clear aligners in orthodontic practice has increased significantly, interproximal reduction is frequently recommended, and is included in the planning of most cases. At the current stage of development of virtual planning, indications for locations in which to perform interproximal reduction do not consider local factors that may interfere with or even contraindicate the procedure, such as sensitivity, the occurrence of previous interproximal reduction and shape/size discrepancy. Interproximal reduction is an invasive and irreversible procedure, and must therefore be planned and executed in such a way as to re-anatomise the crowns, giving them macro- and microscopic characteristics similar to natural ones. The present article aims to shed light on some of the most important factors related to the use of interproximal reduction in clinical practice, especially when associated with clear aligner treatment, so that the treatment goals can be achieved with minimal risk of harm. [ABSTRACT FROM AUTHOR]

This article explores the evolution of mammalian teeth, jaw joints, and middle ear structures. It presents new evidence from fossil studies that sheds light on the earliest tooth diversification of mammaliaforms and the transformation of the reptilian jaw joint into the middle ear apparatus of mammals. The findings challenge previous understandings of tooth patterns and phylogenetic relationships among mammaliaforms. The article discusses the development of the dual jaw joint and the transition to the middle ear ossicles in mammals, as well as the role of Meckel's cartilage in this process. Overall, the available fossil evidence supports the gradual evolution from the dual jaw joint to the separate hearing and chewing apparatuses in mammals. [Extracted from the article]

Alveolar bone, as tooth-supporting bone for mastication, is sensitive to occlusal force. However, the mechanism of alveolar bone loss after losing occlusal force remains unclear. Here, we performed single-cell RNA sequencing of nonhematopoietic (CD45–) cells in mouse alveolar bone after removing the occlusal force. Mesenchymal stromal cells (MSCs) and endothelial cell (EC) subsets were significantly decreased in frequency, as confirmed by immunofluorescence and flow cytometry. The osteogenic and proangiogenic abilities of MSCs were impaired, and the expression of mechanotransducers yes associated protein 1 (Yap) and WW domain containing transcription regulator 1 (Taz) in MSCs decreased. Conditional deletion of Yap and Taz from LepR+ cells, which are enriched in MSCs that are important for adult bone homeostasis, significantly decreased alveolar bone mass and resisted any further changes in bone mass induced by occlusal force changes. Interestingly, LepR-Cre; Yapf/f; Tazf/f mice showed a decrease in CD31hi endomucin (Emcn)hi endothelium, and the expression of some EC-derived signals acting on osteoblastic cells was inhibited in alveolar bone. Mechanistically, conditional deletion of Yap and Taz in LepR+ cells inhibited the secretion of pleiotrophin (Ptn), which impaired the proangiogenic capacity of LepR+ cells. Knockdown in MSC-derived Ptn repressed human umbilical vein EC tube formation in vitro. More important, administration of recombinant PTN locally recovered the frequency of CD31hiEmcnhi endothelium and rescued the low bone mass phenotype of LepR-Cre; Yapf/f; Tazf/f mice. Taken together, these findings suggest that occlusal force governs MSC-regulated endothelium to maintain alveolar bone homeostasis through the Yap/Taz/Ptn axis, providing a reference for further understanding of the relationship between dysfunction and bone homeostasis. [ABSTRACT FROM AUTHOR]

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 (1874), Carey (1920), and Blechschmidt (1948). 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., ((c) 2006 Wiley-Liss, Inc.)

In root cementum of teeth, alternating dark and light lines become visible in cross-sections under the light microscope. These lines bear an apparent resemblance to the annual rings of trees. Numerous studies have been done to correlate the number of cementum lines with the dental age by examining a great number of teeth of known age. Our study used a different approach. If lines in root cementum develop in an annual rhythm and are thus comparable to annual rings of trees, the same or at least a very similar number of these structures should be found in all areas of the root cementum of the same tooth. We counted cementum lines in the buccal, lingual, distal and mesial region of different sections, all from the middle third of the same root. This was repeated in eight teeth. To our surprise, we had immense difficulty in counting reproducible line numbers in the same cementum area at repeated counts. Nevertheless, the same tooth was found to differ markedly in the number of lines in different sections as well as in different regions of the same sections. These differences cannot be ascribed to variations caused by difficulties with reproducible line counting. Therefore, we are more than skeptical about the reliability of counting lines in root cementum as a method for determining the age of human teeth.

Establishing and maintaining mature bone at the bone-device interface is critical to the long-term success of prosthesis. Poor cell adhesion to orthopaedic and dental implants results in implant failure. Considerable effort has been devoted to alter the surface characteristics of these biomaterials in order to improve the initial interlocking of the device and skeleton. We investigated the effect of surface chemistry modification of titanium alloy (Ti-6Al-4V) with zinc, magnesium or alkoxide-derived hydroxy carbonate apatite (CHAP) on the regulation of key intracellular signalling proteins in human bone-derived cells (HBDC) cultured on these modified Ti-6Al-4V surfaces. Western blotting demonstrated that modifying Ti-6Al-4V with CHAP or Mg results in modulation of key intracellular signalling proteins. We showed an enhanced activation of Shc, a common point of integration between integrins and the Ras/Mapkinase pathway. Mapkinase pathway was also upregulated, suggesting its role in mediating osteoblastic cell interactions with biomaterials. The signalling pathway involving c-fos (member of the activated protein-1) was also shown to be upregulated in osteoblasts cultured on the Mg and CHAP modified Ti-6Al-4V. Thus surface modification with CHAP or Mg may contribute to successful osteoblast function and differentiation at the skeletal tissue-device interface.

Aim: Lengthening the mandible by distraction osteogenesis (DO) is nowadays a well recognized technique in maxillofacial surgery. In this study growth factor expression profiles were examined in biopsies taken from six patients undergoing mandibular DO and compared with findings from a sheep model for mandibular DO., Study Design: In all patients (and sheep), the ascending ramus was distracted 10-15 mm at a rate of 1mm/day using an intraoral device. Biopsies were taken from the centre of the distraction zone 21 days after completion of distraction. Using standard immunohistochemical techniques, samples were stained for platelet-derived growth factor (PDGF), transforming growth factor beta (TGF-beta), basic fibroblast growth factor (bFGF) and bone morphogenetic proteins-2, -4 and -7 (BMP-2, -4, -7), matrix metalloproteinases-1 and -3 (MMP-1, -3), the vascular endothelial growth factor (VEGF), a marker for endothelial cells (CD-31) and type IV collagen (Col IV)., Results: Positive staining for PDGF, bFGF, TGF-beta, BMP-2, -4, and -7 was noted in cells and matrix components. There was intense staining for MMP-1. Strong staining for CD-31 and COL IV was observed adjacent to vessels. VEGF staining was less specific. Similar findings were noted in the sheep model., Conclusion: Growth factor expression in the human distraction site is similar to that in the sheep model.