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Ultrastructural localisation of proteoglycans in the odontoblast-predentin region of rat incisor.

The localization of proteoglycans in the predentin of the rat incisor was investigated by ultrastructural histochemistry. Ruthenium red stained the cell coat of the odontoblasts as well as intracellular vesicles. There was also a staining of the extracellular matrix, but not of collagen fibers in the predentin. Treatment with the enzyme hyaluronidase prior to staining with ruthenium red abolished the staining of the vesicles and the extracellular matrix but not that of the cell coat. Bismuth nitrate and phosphotungstic acid gave similar staining of odontoblast vesicles and extracellular matrix. It is likely that the stained structures contain proteoglycans. The importance of these proteoglycans and their ultrastructural localization are discussed in relation to intracellular transport and the calcification process.

Animals↗

In vitro mineralization of a three-dimensional collagen matrix by human dental pulp cells in the presence of chondroitin sulphate.

These matrices were used as cell culture substrates to investigate the influence of extracellular molecules on mineralization. Pulp cells seeded in type I collagen or type I collagen-chondroitin-4-sulphate sponges were able to grow and were morphologically similar to cells responsible for reparative dentine formation in vivo. In sponges consisting of collagen only, the cells elaborated an abundant new matrix which became organized with time and consisted of collagen fibres surrounded by fibrillar material, but no mineralization was observed. In collagen-chondroitin sulphate sponges, cells deposited less and poorly organized matrix; in these, calcification occurred, increasing with time, and at the ultrastructural level, small needle-like crystals containing calcium and phosphorus were scattered throughout the sponge fibres. These observations suggest that chondroitin sulphate might influence in vitro calcification induced by pulp cells.

Cells, Cultured↗

Iron overloading inhibits dentine mineralization.

The present study reveals the inhibitory effect of iron intoxication on the process of dentine mineralization. Wistar rats were injected intraperitoneally with iron dextran at 0.88 g/kg body weight per day for 10 days during the period of odontogenesis. An age-matched group was injected intraperitoneally with bisodium etidronate (EHBP) at 20 mg/kg body weight per day for 10 days. Another age-matched group was treated with similar amounts of saline intraperitoneally and considered as control. At the end of the experimental period the animals intoxicated with iron exhibited non-mineralized dentine and mineralized bone. The animals treated with EHBP showed non-mineralized dentine and bone. These findings would suggest the existence of different mineralization mechanisms for bone and dentine.

Alveolar Process↗

Expression of osteocalcin in cementoblasts forming acellular cementum.

To determine the phenotypic expression of cementoblasts responsible for acellular cementum, an immunohistochemical study was performed using a polyclonal antibody raised against the aminoterminal peptide of rat osteocalcin (OC). Maxillary first molars of Wistar male rats aged 2 and 3 wk were used for observations. Serial sections of decalcified paraffin embedded specimens were stained either with hematoxylin and eosin or with the anti-OC antibody. In 2-wk-old rats, apical roots were lined with the epithelial root sheath. A thin layer of acellular cementum was seen at most of the root surface, but was not seen near to root apex. In 3-wk-old rats, cellular cementum began to be formed at root apex, and acellular cementum became more thick than in 2-wk-old rats. Acellular and cellular cementum were lined with the fibroblast-like cells. Osteocalcin staining was detected in cells lining root surface in both 2- and 3-wk-old rats. Almost all cells lining cellular cementum were positive for OC. In contrast OC positive cells lining acellular cementum and root surface devoid of cementum appeared at a specific site of the root. The cells at the interradicular area of root surface were positive but the cells at the outer area (the opposite side of the interradicular area) were negative for OC. Osteoblasts and odontoblasts were positive with the antibody. The present results suggest that the OC expression of cementoblasts forming acellular cementum is similar to that of cells forming cellular cementum as well as osteoblasts and odontoblasts, and has a role for calcification of acellular cementum.

Animals↗

[Relationships between mandibular canine calcification stages and skeletal maturity].

OBJECTIVE: To study the relationship between mandibular canine calcification and skeletal maturity. METHODS: Hand wrist and mandibular canine radiographs of 106 boys and 152 girls in Beijing area were assessed. RESULTS: Mandibular canine calcification coincides with the initiation of puberty in male (93.8%) and with the deceleration of puberty in female(89.7%). However the coincidence with the indicators of other pubertal stages was less than 50%. CONCLUSION: Mandibular canine calcification just can give an initial assessment for estimating the timing of puberty.

Adolescent↗

Ca-ATPase and ALPase activities at the initial calcification sites of dentin and enamel in the rat incisor.

Enzymatic activities of calcium-magnesium dependent adenosine triphosphatase (Ca-ATPase) and nonspecific alkaline phosphatase (ALPase) were localized at the initial calcification sites of dentin and enamel of rat incisor teeth using electron-microscopic cytochemistry. Ca-ATPase was localized in the Golgi cisternae, cytoplasmic vesicles and along the outer surface of the presecretory and secretory ameloblasts, whereas it was totally absent from the odontoblasts in the pulp. Inversely, ALPase reaction was localized along the outer surface of the odontoblasts, but almost completely absent from the ameloblasts. Diffuse extracellular reactions of both enzymes were distributed throughout the unmineralized fibrous matrix of mantle dentin in which a large number of matrix vesicles were scattered. Both Ca-ATPase and ALPase reactions, which appeared in the matrix vesicles in the process of formation of mantle dentin, became most conspicuous at the site of initial dentin calcification. At this stage, an intense Ca-ATPase reaction also appeared along some of the collagen fibrils adjacent to the reactive matrix vesicles. No ALPase reaction was localized along these Ca-ATPase reactive collagen fibrils. Our observations suggest strongly that Ca-ATPase in the matrix vesicles originates from the inner enamel epithelium and/or preameloblasts whereas ALPase originates from the odontoblasts in the pulp. The importance of the coexistence of both enzymes for the control of initial calcification of dental hard tissues is suggested.

Alkaline Phosphatase↗

Delayed calcification and congenitally missing teeth. Case report.

A case of delayed differentiation and/or calcification, that is, development, followed by accelerated growth of the maxillary second premolars is presented. This condition was accompanied by the congenital absence of mandibular second premolars even though maxillary and mandibular third molar germs were present. The case was followed over a five-year period. It seems that agenesis of any tooth might be found in conjunction with delay in development of any other tooth. Moreover, full development of a tooth germ (followed by an accelerated growth) as well as pathological changes may occur, despite an initial marked delay in tooth development.

Anodontia↗

Phosphopeptides of enamel matrix.

Although the tripeptides Glu-O-Phosphoserine-Tyr and Glu-O-Phosphoserine-Leu have been identified in embryonic bovine enamel proteins, 1, 2 the issue of whether both sequences occur in each of the phosphopeptides, or whether certain sequences occur in specific peptides only, has recently been resolved by isolating homogeneous samples of E33 and E44. All three of the Ser residues of both peptides are phosphorylated. All three in E3 are in the sequence Glu-O-Phosphoserine-Leu, and all three in E4 are in the sequence Glu-O-Phosphoserine-Tyr. It was not possible to sequence either of the polypeptide chains directly by automatic peptide sequencing. However, a partial sequence of E4 was constructed from data derived from peptides isolated after cyanogen bromide, trypsin and chymotrypsin digestions. The presence of Glu, Tyr and Leu adjacent to and near the O-Phosphoserine [Ser(L)] residues and the 2 degrees, 3 degrees and higher ordered structures of the enamel phosphopeptides may be important in calcium binding and mineralization.

Amino Acid Sequence↗

Dysplastic enamel in odontoma: a light microscopic, microradiographic and SEM study.

Dysplastic enamel and calcifications at the enamel surface in 7 odontomas were studied, using correlated light microscopy of decalcified and undecalcified material, microradiography and SEM. Much of the dentin in the odontomas was not covered with enamel. When present, the enamel was immature and assumed a prismatic structure. The prisms were irregular distributed and associated with spherical calcifications. The calcifications adhering to the enamel surface or separated from it presented variations in size, morphology, staining reactions and radiodensity. The correlated techniques of light microscopy, microradiography and SEM indicated that all the calcifications adhering to the enamel surface and part of those separated from it may be related to a pathological process of amelogenesis. Most of the calcifications separated from enamel and often formed around nidi of ghost cells, are the result of a dystrophic mineralizing process, definitely distinct from amelogenesis.

Calcinosis↗

Transmission electron microscopy of reparative dentin in rat molar pulps. Primary mineralization via extracellular matrix vesicles.

A reparative dentin bridge was induced in rat molar teeth by pulp exposure and capping with calcium hydroxide. Transmission electron-microscopic examination after 10 days revealed the presence of odontoblastic cells and collagenous matrix with focal calcifications. The calcifying fronts were composed of hydroxyapatite crystals. Numerous extracellular matrix vesicles were scattered between the forming cells and the calcifying fronts. Some of the vesicles contained electron-dense material and in others, apatite crystals were detected. Matrix vesicles could not be identified in normal, mature calcifying dentin matrix. In view of the present observations and studies on surgical manipulations in articular cartilage, it is concluded that matrix vesicle calcification may result from alterations in the metabolic state of mesenchymal tissues. These changes can be induced surgically or chemically.

Animals↗