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Characterization of glycosaminoglycans during tooth development and mineralization in the axolotl, Ambystoma mexicanum.

Glycosaminoglycans (GAGs) involved in the formation of the teeth of Ambystoma mexicanum were located and characterized with the cuprolinic blue (CB) staining method and transmission electron microscopy (TEM). Glycosaminoglycan-cuprolinic blue precipitates (GAGCB) were found in different compartments of the mineralizing tissue. Various populations of elongated GAGCB could be discriminated both according to their size and their preferential distribution in the extracellular matrix (ECM). GAGCB populations that differ in their composition could be attributed not only to the compartments of the ECM but also to different zones and to different tooth types (early-larval and transformed). Larger precipitates were only observed within the dentine matrix of the shaft of the early-larval tooth. The composition of the populations differed significantly between the regions of the transformed tooth: pedicel, shaft and dividing zone. In later stages of tooth formation, small-sized GAGCBs were seen as intracellular deposits in the ameloblasts. It is concluded that the composition of GAGCB populations seems to play a role in the mineralization processes during tooth development in A. mexicanum and influence qualitative characteristics of the mineral in different tooth types and zones, and it is suggested that GAGs might be resorbed by the enamel epithelium during the late phase of enamel formation.

Ambystoma mexicanum↗

The management of postbleaching cervical resorption.

Cervical resorption has been repeatedly associated with intracoronal bleaching procedures. This report reviews literature associated with postbleaching resorption, a case, and a rationale for management. We observed cervical resorption upon recall of an intracoronal bleaching case. It was successfully treated with a calcium hydroxide "recalcification" procedure. Twenty-three months later, no further resorption has been observed.

Adolescent↗

Effect of retinoic acid on osteopontin expression in rat clonal dental pulp cells.

We studied the effect of retinoic acid on osteopontin synthesis and the mRNA expression in rat clonal dental pulp cells, RPC-C2A. An immunoprecipitation assay clarified that retinoic acid caused an increase in phosphorylated osteopontin synthesis that was dose-dependent, and marked increases were observed at retinoic acid concentrations of 10(-6) to 10(-5) M (1.7-fold). A Northern blotting analysis revealed a similar pattern of increase in osteopontin mRNA expression of up to 6.2-fold of control levels. Because osteopontin has an important role in the mineralization process, these results suggest that retinoic acid regulates mineralization, which takes place in the pulp cavity, including reparative dentin formation.

Animals↗

[Problems raised by the use of tetracyclines in the first years of life].

Tetracyclines administered during the first years of life may produce serious unwanted side-effects, such as intracranial hypertension and, chiefly, abnormalities of the teeth and bones. The tetracyclines have a special affinity for certain metal cations, notably calcium, with which they form relatively stable complexes. When given to children or pregnant women they stain the deciduous and permanent teeth a lasting yellowish-brown or give rise to dental abnormalities; they may also slow down skeletal growth. For these reasons tetracyclines should not be prescribed to children and pregnant women unless absolutely necessary, and other classes of antibiotics should be preferred whenever possible.

Age Factors↗

In vivo study of the pulp reaction to Fuji IX, a glass ionomer cement.

OBJECTIVE: Our aims were to investigate the pulp biocompatibility of Fuji IX, a glass ionomer cement (GIC) used as a restorative material in cavities prepared in rat's upper molars, and to assess the value of this in vivo model for testing dental biomaterials. METHOD: Half-moon class V-like cavities were drilled on the mesial aspect of 26 rat upper first molars. Half of the experimental rats whose molars were restored with the GIC were killed after 8days and the second half after 30days. They were compared with two control groups, also submitted to cavity preparation, but with cavities left unfilled. Again, half of the control rats were killed at 8 days and the second half after 30days. Following intracardiac perfusion with the fixative solution, the specimens were processed to histologic procedures. RESULTS: After 8 days, in both groups a few inflammatory cells were observed. The odontoblastic layer was disrupted and dilated blood vessels were seen in the pulp area related to the cut tubules. The experimental group displayed a moderate inflammatory reaction whereas only a slight reaction was detected in the control group. In few teeth, bacteria were visualized in dentine tubules beneath the GIC restoration. Such colonies were not observed in unfilled molars.After 30days, in both groups, the pulp tissue recovered and displayed a normal appearance. Disruptions of the odontoblast layer were not visible anymore. Bacteria penetration into dentine tubules was reduced compared with the 8-day situation. A thick layer of reparative osteodentine was formed. However no difference in thickness was detected between the experimental and control groups, supporting that the formation of reparative dentine is not impaired. Irregular mineralizations including calcospherites were induced by the GIC. CONCLUSIONS: This study demonstrates that, despite small alterations in the mineralization processes, the GIC Fuji IX has a good biocompatibility and does not induce any harmful effect on pulp cells.

Animals↗

Craniofacial structure related to inheritance pattern in amelogenesis imperfecta.

The aim of this study was to investigate the craniofacial structure in 66 children and adolescents, 34 girls and 32 boys, with known clinical manifestations and inheritance patterns for amelogenesis imperfecta (AI), and to compare the results with those obtained in a control group of age and sex matched persons with normal occlusion. The ages ranged from 6.8 to 21.2 years. Clinically, AI was divided into cases characterized by either hypoplasia or hypomineralization of the enamel. In a further subgrouping, eight clinical variants were diagnosed. Measurements of 12 angular and 15 linear, parameters on lateral cephalometric radiographs were included in comparisons between the AI and the control group. Compared with the control group, the AI group displayed statistically significant differences indicating a skeletal open bite. In the analysis of inheritance patterns and clinical manifestations, a skeletal open bite was associated with autosomal dominant (AD) and X-linked inheritance, and in the AD group with hypomineralization. When all cases except those with X-linked inheritance were pooled, deviations indicating a skeletal open bite were found in the subgroups "rough hypoplastic AI" and "hypomineralization AI." Since a skeletal open bite was found both with X-linked inheritance and in some of the subgroups connected with autosomal inheritance, the hypothesis of a pleiotropic gene effect as the cause of the simultaneous occurrence can be ruled out. The influence of modifying genes or environmental factors is suggested.

Adolescent↗

Immunochemical and biochemical characteristics of enamel proteins in hypocalcified amelogenesis imperfecta.

Amelogenesis imperfecta is a hereditary disease of the enamel that is unassociated with generalized defects. Cases of the condition are clinically classified into three groups: hypoplastic, hypomaturation, and hypocalcified. In this study, soluble protein fractions of the enamel from three patients with hypocalcified amelogenesis imperfecta were examined through the use of immunochemical and biochemical techniques. In immunochemical analyses done with a polyclonal anti-amelogenin antibody, all samples from enamel in which there was amelogenesis imperfecta were found to contain considerable amounts of amelogenin peptides. When an enamel sample from one patient was examined by Western-blot transfer and immunobinding analysis, the amelogenin fraction was found to consist of a 26-kDa molecule thought to be normally present in the outer layer of secretory-stage enamel. This enamel was also found to contain albumin as one of the major constituents of the protein fraction. These results suggest that hypocalcified amelogenesis imperfecta may in part be caused by a disturbance in matrix protein degradation during the maturation phase.

Adolescent↗

Comparative study on the chronology of third molar mineralization in a Japanese and a German population.

In Germany, a sharp increase in forensic age estimations of living persons has been observed in recent years. German law defines four legally relevant age limits: 14, 16, 18 and 21 years. In these age groups, radiographic assessment of the mineralization status of third molars is of particular importance. So far, the influence of ethnicity on the mineralization rate has been insufficiently analyzed. A total of 3031 orthopantomograms of 1597 Japanese and 1434 Germans aged between 12 and 26 years were examined. The mineralization status of third molars was evaluated on the basis of the classification proposed by Demirjian. For the individual mineralization stages, the study presents the mean values and standard deviations (SD) separately for both populations and sexes. The majority of probands from both the Japanese and the German population achieved the C stage and the late G and H stages of third molar development at similar ages. Significant differences between Japanese and Germans were observed, however, with regard to the D, E and F stages defined by Demirjian. Japanese men and women achieved the D, E and F stages approximately 2-3 years later than German men and women. In addition to forensic age determination in living persons, the presented reference data can also be used for age estimations of unidentified deceased persons and skeletons.

Adolescent↗

On the origin of intrinsic matrix of acellular extrinsic fiber cementum: studies on growing cementum pearls of normal and bisphosphonate-affected guinea pig molars.

Cementum pearls (CPs) belong to a type of acellular extrinsic fiber cementum (AEFC) that form on the maturing enamel of guinea pig molars. This study aimed to elucidate the forming process of intrinsic matrix of AEFC using the CPs of normal and bisphosphonate-affected guinea pig molars as experimental models. A group of guinea pigs were subjected to continuous administration of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) for 2 wk to inhibit mineralization of growing CPs. Fenestration of the enamel organ and migration of periodontal cells on to the exposed surface of maturing enamel appeared to be unaffected by HEBP, whereas de novo formation as well as growth of pre-existing CPs did not proceed under the same conditions. Immunoreactions for osteopontin were located exclusively on the mineralized matrix of preformed CPs, implying the absence of additional deposition or accumulation of putative intrinsic cementum matrix on the affected CPs, where the propagation of mineral phase had been arrested. In both normal and HEBP-treated groups, distinct enzymatic reactions for alkaline phosphatase appeared on the cells of the periodontal ligament associated closely with the sites of CP formation, and along the mineralization front of CPs. These observations suggest that the mineralization process per se plays a central role in the deposition of AEFC matrix and that alkaline phosphatase of periodontal cells penetrating through the enamel organ to the maturing enamel surface plays a key role in the mineralization process of CPs.

Alkaline Phosphatase↗

Enamel proteins and extracellular matrix molecules are co-localized in the pseudocystic stromal space of adenomatoid odontogenic tumor.

In order to examine the functional differentiation of tumor cells of adenomatoid odontogenic tumor (AOT) as ameloblasts and to determine the participation of the extracellular matrix (ECM) in the formation of its characteristic histologic architecture, tissue samples from five cases of adenomatoid odontogenic tumor were examined by immunohistochemical staining for enamel proteins and ECM molecules. Amelogenin, enamelin, laminin, heparan sulfate proteoglycan, fibronectin, collagen type IV and type V were immunolocalized within the luminal space and along the inner rim of duct-like structures. Eosinophilic hyaline droplets within the whorled or rosette masses of tumor cells showed basically the same staining pattern as the luminal contents. High columnar tumor cells that formed duct-like structures were immunopositive for amelogenin, while the staining intensity decreased with flattening of the cells, which was a result of luminal growth. The findings suggest that the constituent cells of duct-like structures are differentiated once to ameloblasts but fail to mature further due instead to increased production of ECM molecules and due to their retention in the lumina. It is possible to regard these special structures in AOT as stromal pseudocysts.

Amelogenin↗

Distribution of non-collagenous dentin matrix proteins and proteoglycans, and their relation to calcium accumulation in bisphosphonate-affected rat incisors.

It has been reported that multiple injections of 1-hydroxyethylidene- 1,1-bisphosphonate (HEBP) to rats prevent mineralization of incisor dentin, thereby revealing high concentrations of calcium in the non-mineralized matrix of circumpulpal dentin. To identify the molecules responsible for calcium accumulation in circumpulpal dentin matrix, rats were injected daily with HEBP (8 mg P/kg) for 7 d, and the incisors processed for various histochemical and immunohistochemical staining of non-collagenous matrices of dentin. Cuprolinic blue reactions for proteoglycans (PGs) were equally distributed in non-mineralized matrix of mantle and circumpulpal dentin layers. Dentin sialoprotein (DSP) and osteopontin (OPN) immunoreactions were found in non-mineralized circumpulpal dentin matrix, but not in mantle dentin. In normal incisors, however, predentin matrix showing significant DSP immunoreactivity was negative for Ca-GBHA reactions. HEBP-affected, non-mineralized OPN immunopositive bone matrix was also non-reactive for calcium. From these observations, neither PGs, OPN nor DSP appear to be responsible for calcium accumulation in HEBP-affected circumpulpal dentin. Stains-all reactive component, possibly dentin phosphoprotein (DPP), only showed the same distribution as that of Ca-GBHA in both HEBP-affected and normal dentin matrix, implicating a possible contribution of DPP to calcium accumulation in circumpulpal dentin and, hence, to appositional mineralization of dentin.

Animals↗

Exclusion of candidate genes in two families with autosomal dominant hypocalcified amelogenesis imperfecta.

The amelogenesis imperfectas (AI) are a group of hereditary enamel defects characterized by clinical and genetic diversity. The most common AI types are inherited as autosomal traits. Three mutations of the enamelin (ENAM) gene have been found in cases of autosomal dominant hypoplastic AI. The gene(s) responsible for hypocalcified forms of AI have not been identified, although a number of autosomal genes have been proposed as candidates for AI based on their expression by ameloblasts, including ameloblastin and enamelin (chromosome 4q13.3), tuftelin (chromosome 1q21), enamelysin (chromosome 11q22.3-q23) and kallikrein 4 (chromosome 19q13.3-q13.4). To localize the gene(s) responsible for autosomal dominant hypocalcified AI, we evaluated support for/against linkage of AI to genetic markers spanning five AI candidate genes in two extended families. Our data excluded all proposed candidate gene regions as causal for autosomal dominant hypocalcified AI in these families. These linkage findings provide further evidence for genetic heterogeneity among families with autosomal dominant AI and indicate that, at least, some forms of autosomal dominant hypocalcified AI are not caused by a gene in the five most commonly reported AI candidate genes.

Amelogenesis Imperfecta↗

3D X-ray microscopic study of the extent of variations in enamel density in first permanent molars with idiopathic enamel hypomineralisation.

OBJECTIVE: To measure mineral concentration distributions within teeth with idiopathic enamel hypomineralisation, a condition in which developmental defects are seen in first permanent molars, and/or incisors. DESIGN: X-ray microtomographic and 3D x-ray microscopy. SETTING: UK University, 2001. MATERIALS AND METHODS: X-ray microtomographic measurements of the extent of hypomineralisation in two affected molars and two contralateral controls extracted from the same patient. RESULTS: The control molars were visibly normal. The affected molars showed hypomineralised yellow opaque enamel with regions of breakdown. X-ray microtomographic images showed; a 20% reduction in mineral concentration of affected enamel (most cases involved full enamel thickness); hypomineralised enamel had a mineral concentration gradient opposite to that of normal enamel; regions of hypomineralisation distributed randomly throughout affected teeth, (apart from cervical region which was less severely affected). CONCLUSIONS: The pattern of mineral concentration suggests a disturbance during the maturation process. Differences in susceptibility of the ameloblasts during different stages of dental development may explain the asymmetric distribution of the defects. Topical fluoride applications may help promote post eruption maturation of the surface layer in these teeth. The use of fissure sealants and adhesive materials appears to prevent further breakdown.

Child↗