[The role and possibilities of an original calcium therapy in dentistry].
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The bone, dentin, and cementum of the mature individual are comprised from a dense collagenous fiber network into which the carbonate-apatite mineral phase is deposited. It is hypothesized that a set of collagen-interactive acidic phosphoproteins are secreted by the osteoblasts, odontoblasts, and cementoblasts into the preformed collagenous matrix. These proteins then interact specifically with the collagen and nucleate apatite formation on and within the fibrils. These phosphoproteins may also regulate the morphology, rate of growth, and stability of the mineral phase crystals. The acidic matrix phosphoproteins may thus be considered as the crucial regulators of mineralization and tissue stability. In the dentin system, these regulatory proteins are synthesized, posttranslationally modified, and secreted in vesicles different from the collagen secretory vesicles. Mineralization occurs as the regulatory proteins are deposited on the preformed fibrils. This model requires testing in the bone system. In dentin, in the absence of tissue turnover, the resident phosphoproteins are degraded in situ over time, perhaps changing the properties of the tissue. Regulation of synthesis, secretory pathways and retention of integrity within the matrix are thus important areas for further investigation.
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Bovin dentin, bone and tendon slices, and rat bone, readily mineralize to variable degrees after demineralization by (EDTA) at pH 7.4, but they fail to mineralize after dimeralzation with acetic acid (HAc) at pH3.0. The demineralized dentin, but neither bone nor tendon, contained organically bound phosphate. The EDTA-demineralized dentin contained less phosphate than HAc-demineralized dentin. HAc-demineralized rat dentin contained high levels of phosphate. Since the EDTA- and HAc-demineralized rat dentin contained widely different levels of phosphate, yet both mineralized, it was concluded that phosphoprotein had little effect on nucleation. The reason why HAc-demineralized tissue other than rat dentin failed to nucleate and mineralize was not clarified.
Close organic-inorganic relationships exist in all calcified tissues, the inorganic substance being linked to crystal ghosts (CGs). These are organic, crystal-like structures present in areas of initial calcification. In cartilage and bone, they form aggregates with the same morphology and distribution as the calcification nodules; in enamel, they consist of long filament- and ribbon-like structures, having the same arrangement as untreated crystals. CGs of cartilage and bone are acidic structures with histochemical properties of proteoglycans; CGs of enamel probably correspond to enamelins. The close morphologic similarity between CGs and crystals suggests that the former have a role in the formation of the latter.
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Primary culture of explants of human dental pulp tissue allows the study of the cytophysiology and differentiation of the cultured cells over a two-week period. The distribution of calcium was found in two different experimental conditions : with and without a calcium loading, by mean of a lead technique checked by microprobe analysis. The existence of two cell populations was revealed. Intra-mitochondrial ring-like granules characterize type 1 cells when overloaded, while a strong calcium storage is detected in the rough endoplasmic reticulum, Golgi apparatus and mitochondrial (without any inner organization of the deposits) of the type 2 cells. Our results also show the presence of calcium on gap-junctions (revealed) by lanthanum method), and on the extracellular matrix (collagen fibres and complex carbohydrates). The ability of some mitochondria to store calcium (ring-like granules) suggests that the type 1 cells are fully differentiated in odontoblast-like cells and perhaps engaged in mineralization processes. The calcium binding sites, localized on the extracellular matrix may therefore be considered as the earliest foci of calcification.
The aim of this study was to investigate the relationship between proteoglycans (PGs) and collagen fibrils at the early mineralization process of mantle dentin. Ten first molar dental germs of rats were removed and fixed in glutaraldehyde/formaldehyde in cacodylate buffer and post-fixed in osmium tetroxide. The samples were dehydrated and embedded in epoxy resin. Ultrathin sections were contrasted and analyzed in TEM before and after treatment with EDTA, chondroitinases AC and ABC. After EDTA treatment, a electrondense substance associated with collagen fibril was removed, and did not stain again. A high magnification of these areas showed globular structures with 15 nm diameter surrounding collagen fibrils. In advanced mineralization areas, collagen fibrils showed a banded pattern and at high magnification the fibrils presented a light 10 nm ring inside and a dark 10 nm ring outside. After chondroitinase treatment, the electrondense substance associated with collagen fibrils was removed, showing a banded pattern of clear and dark areas along them. From morphological data, the authors proposed a model of interaction between PGs and collagen fibrils, where glicosaminoglycans chains are inside the fibrils, while the protein core remains outside. That stereochemical arrangement would start the crystal nucleation.
The mineralized matrices of enamel, cementum, dentin, calcified cartilage and bone are similar in their ability to form a microenvironment that facilitates deposition of hydroxyapatite. However, they are not identical, as witnessed by the nature of apatite crystals that are formed. Enamel is devoid of collagen; and is composed of enamelins, amelogenins, tuftelin and ameloblastin, first described at this meeting. Cementum, dentin and bone matrices are composed primarily of type I collagen, however, each matrix may also contain unique moieties. The exact composition of cementum is not fully known, but in dentin there are unique matrix proteins, phosphophoryn (dentin phosphoprotein, DPP), a distinctive dentin matrix protein (DMP-1), and dentin sialoprotein (DSP). In bone, dentin and cementum, the matrix proteins include proteoglycans (versican, decorin, biglycan) and hyaluronan, glycoproteins which are often phosphorylated and sulfated (osteonectin, RGD-containing proteins) and gla-containing proteins (matrix gla protein, protein S, osteocalcin). The exact nature of all the non-collagenous proteins of calcified cartilage is not yet fully known. While there are no definitive functions for any of the mineralized matrix proteins to date, they most likely participate in regulation of cell metabolism, matrix deposition and mineralization, and bone turnover.
When calcifying tissues were extracted with hot pyridine or hot benzene and then decalcified and stained with Sudan black B, the areas where mineralization was being initiated stained strongly, the rest of the calcified tissues being unstained. Histochemical methods showed that lipids were responsible for the staining. They were isolated biochemically and found to be phospholipids, very resistant to extraction before decalcification of the tissues and consisting predominantly of phosphatidyl serine and phosphatidyl inositol. It was proposed that these phospholipids were active at nucleating sites in apatite crystal formation, since it is known that phosphatidyl serine binds calcium strongly. It has been shown that phosphatidyl serine is present in matrix vesicles.
Chronic calcitonin (CT) deficiency was shown to have several effects on the incisors of young growing rats. The incisors had a significantly wider predentin layer, interglobular dentin, and frequent pulpal exposures at the incisal edge. It was concluded that CT plays a role in the normal calcification of dentin matrix.
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