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Calcium ion activity and pH in the odontoblast-predentin region: ion-selective microelectrode measurements.

Ca2+ ion activities and pH were measured in the odontoblast/predentin region of rat incisors by means of the microelectrode technique. In Ringer solution, the apparent resting membrane potential of odontoblasts was determined to be -24 +/- 4 mV (mean +/- SE), whereas the odontoblast intracellular pH was found to be 6.66 +/- 0.02. The values obtained are within the range of other cell types, as measured in similar incubating solutions. The pH in the extracellular predentin was higher than the intracellular pH, 7.00 +/- 0.02. The Ca2+ ion activity in predentin (pCa = 2.94 +/- 0.15) was found to be significantly (P less than 0.001) higher than that in the dental pulp extracellular fluid (pCa = 3.37 +/- 0.14). The 2-3 times higher calcium activity extracellularly in predentin, compared with the dental pulp, implies the existence of some ion-concentrating mechanism across the odontoblast layer in the direction of the mineralization front.

Animals↗

Qualitative electron probe analysis of secretory ameloblasts and odontoblasts in the rat incisor.

Rapidly frozen growing rat incisors were freeze fractured and freeze dried in preparation for energy dispersive X-ray emission microanalysis in a scanning electron microscope. Ca levels were found to be elevated in the distal cell body of odontoblasts, whereas Ca was uniformly low over all parts of the cell body of secretory ameloblasts. The results suggest fundamental differences in the mechanisms by which these two cell types process Ca, and that Ca possibly diffuses through the secretory ameloblast layer on its way to the enamel.

Ameloblasts↗

Epitaxial overgrowth of apatite crystals on the thin-ribbon precursor at early stages of porcine enamel mineralization.

The aim of the present work was to investigate changes in cross-sectional morphologies of enamel crystallites as a function of location in secretory porcine enamel. Enamel tissues were obtained from 5- to 6-month-old slaughtered piglets. For examination by electron microscopy, a portion of the secretory enamel was embedded in resin and ultrathin sections were prepared with a diamond knife. In parallel studies, compositional and structural changes of enamel mineral were assessed by chemical analysis and Fourier transform infrared (FTIR) spectroscopy. For this purpose, two consecutive layers of the outer secretory enamel, each approximately 30 microns thick, were separated from the labial side of permanent incisors. Using high-resolution electron microscopy, early events of enamel crystal growth were characterized as the epitaxial growth of small apatite units on the lateral surfaces of the initially precipitated thin ribbon. These apatite units had regular triangle or trapezoid cross-sections. After fusions of those isolated trapezoids on both lateral sides of the platy template, the resulting enamel crystallites had the well-documented flattened-hexagonal shapes in cross-sections. The initially precipitated thin plate was buried inside the overgrown apatite lamella and then retained as a central dark line. Similar morphological evidence for the epitaxial nucleation and overgrowth of carbonatoapatite on the platy template was obtained in vitro. Chemical and FTIR analyses of the enamel layer samples showed that the characteristics of the youngest enamel mineral were distinct from those of enamel crystals found in older secretory enamel.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Plasma fluoride and enamel fluorosis.

It is postulated that tissue fluid F concentrations are the primary determinants of flouride effects on bones and developing teeth and that these concentrations are dependent on, or mirrored by, blood plasma F. It has earlier been shown that the plasma F levels are dependent on the dietary F supply as well as on skeletal F concentration. Fasting and post-ingestion or postinjection plasma F levels have been determined in rats on F doses that cause different degrees of enamel fluorosis. The results indicate that temporary peak values rather than elevated fasting values are responsible for the occurrence of enamel fluorosis and that the peak values must approach about 10 muM in order to block enamel formation by the ameloblasts. The diagnostic and prognostic importance of plasma F determinations is discussed.

Amelogenesis↗

Proteoglycans of dentine and predentine.

A density gradient system is presented by which dentine and predentine are separated, leaving an intermediate fraction, which contains material from the mineralization front. From the fractions thus obtained the proteoglycans were extracted with 4 M guanidinium chloride and further purified in urea on a DEAE column. The glycosaminoglycans extracted from dentine appeared to be protein-bound, as judged from papain digestion experiments. The polydispersity of dentine proteoglycans seemed to depend, at least partly, upon the polydispersity of its glycosaminoglycans. The materials extracted from the three tissue fractions were ellted in a similar way from Sepharose 6 B, and the amino acid composition of the preparation was determined. The differing proteoglycan patterns of the three tissue fractions indicate a metabolism related to the mineralization front.

Amino Acids↗

Effects of fluoride, calcium, and phosphate administration on mineralization in rats.

Seven days before a fluoride injection of 20 mg sodium fluoride per kg body weight, 3-month-old rats grown on a standard pellet diet containing 0.8% calcium and 1.4% phosphate were given a diet of rice with only 0.025% calcium and 0.1% phosphate. Microradiographs of the continuously growing incisors showed a hypermineralized and subsequent hypomineralized zone. Blood analysis demonstrated a decrease and a subsequent reestablishment of plasma calcium concentration. In some experiments calcium and phosphate were administered to compensate the hypocalcemia which prevented the hypomineralized zone from arising. A delay of calcium and phosphate administration led to formation of a mineralized band within the hypomineralized zone. The results are discussed with reference to calcium homeostasis.

Administration, Oral↗

Variations in the composition of developing rat incisor enamel.

The developing enamel of rat incisors was dissected into a series of samples extending from the newly-formed partially-mineralised matrix to the mature.enamel. Chemical analysis showed that, on a dry weight basis, the tissue achieved the composition of mature enamel well before the completion of mineral uptake. The enamel at this stage was porous and relatively soft. As more mineral was acquired, its hardness increased. Throughout the developing region, the Ca:P ratio remained fairly constant, but the CO2:P and Mg:P ratios both decreased due, apparently, to dilution by an influx of relatively C02- and Mg-free mineral.

Amelogenesis↗

Effect of matrix bound phosphate and fluoride on mineralization of dentin.

Completely demineralized bovine dentin mineralized to a high degree, but only after extraction of about half of its organic phosphate. Initiation of mineral formation depended on the concentrations of Ca2 plus, Pi, HCO3 minus and H plus ions in the incubation solution. After nucleation, mineral accretion by the dentin continued in solutions that did not, themselves, initiate mineralization. The nucleation phase, but not the subsequent accretion of mineral, was sensitive to small changes in temperature. Inclusion of 0.05 mM NaF in the solution markedly reduced the amount of mineral deposited during the nucleation phase but apparently enhanced subsequent mineralization.

Acetates↗

Remineralization of dentin in vitro.

The remineralization of completely demineralized bovine dentin was studied in vitro by monitoring the velocities of fall of small particles through calcifying solutions. The density of a particle of dentin may be found from its velocity of fall through a fluid using Stokes' law. The minimum concentration product of calcium and acid phosphate ions of the solution in which remineralization would take place was 3.6 (mM)2 in the presence of 22 mM bicarbonate, pH 7.35, and ionic strength 0.1. This just above the solubility product of brushite (CaHPO4-2H2O). Incubation of decalcified dentin in a phosphoprotein removed from dentin during demineralization, or in phosvitin, had no effect on remineralization. The rates of remineralization and of the fraction remineralized were inversely proportional to particle size. This inverse correlation may be due to deposition of mineral in a surface layer of constant depth irrespective of particle size. The fraction of a particle remineralized was greatly increased by the use of highly supersaturated calcifying solutions or by the incorporation of fluoride into the solutions. The empirical reaction order of remineralization for both calcium and phosphate ions was found to be unity, which is, within the error limits, equal to the order of growth of seed crystals of hydroxyapatite in calcifying solutions of the same composition.

Animals↗

The secretory stage of amelogenesis in rat mandibular incisor teeth observed by scanning electron microscopy.

In the scanning electron microscope (SEM), the structural organization of rat incisor enamel was divided into five regions: the initial prismless layer at the dentin-enamel junction; the decussating rods of inner enamel; intermediate enamel, where the structure changes from inner to outer enamel; outer enamel with parallel rods; and the surface prismless layer. There were alterations in the orientation of the ameloblasts and characteristic changes in the morphology of the Tomes' processes during the secretion of each of these regions. Initially, the developing Tomes' processes were seen within shallow pits which also contained fibrous material from dentin matrix. During the deposition of inner enamel, alternate rows of Tomes' processes were held at right angles to each other by cytoplasmic projections which originated from the proximal regions of the Tomes' processes. The cytoplasmic projections reorganized to perpendicular orientation relative to the enamel surface during the synthesis of outer enamel. Tomes' processes of outer enamel appeared as elongated flaps extending parallel to the long axis of the tooth. In the late secretory stage, the microvillous projections disappeared and the Tomes' processes were withdrawn into button-shaped structures which coalesced to form a smooth surface that rested on the enamel.

Ameloblasts↗

Changes in heated and in laser-irradiated human tooth enamel and their probable effects on solubility.

Enamel of intact human teeth laser irradiated in vitro under certain conditions is known to have less subsurface demineralization than unirradiated enamel on exposure to acid; consequently, the potential use of laser irradiance to reduce caries is apparent. The laser-induced physical and/or chemical changes that cause this reduced subsurface demineralization are not known. A laser-irradiated tooth enamel surface will have a temperature gradient that decreases towards the dentin junction. Dependent on irradiant conditions, the temperature may range from greater than 1400 degrees C at the surface to near normal at the dentin-pulp junction. Along this steep temperature gradient, different compositional, structural, and phase changes in the tooth enamel are to be expected. Identification of changes occurring along this gradient has bearing on understanding the dissolution reduction mechanism and, in turn, optimizing its effect. Changes in laser-irradiated material from the highest temperature region have been characterized, but those occurring in sequential layers of decreasing temperatures have not. Since the laser-induced changes are expected to primarily arise from localized heating, previously reported thermally induced changes in tooth enamel on heating in conventional furnaces were utilized to infer corollary changes along the gradient in laser-irradiated tooth enamel. These thermally inferred changes which resulted in modifications in the tooth enamel apatite and/or newly formed phases were correlated with their probable effects on altering solubility. A temperature gradient range from 100-1600 degrees C was considered with subdivisions as follows: I, 100-650 degrees C; II, 650-1100 degrees C; and III, greater than 1100 degrees C. Two of the products formed in range III, alpha-Ca3(PO4)2 and Ca4(PO4)2O, and also identified in the fused-melted material from laser-irradiated tooth enamel, are expected to markedly increase solubility in those regions that contain considerable amounts of these compounds. Products and changes occurring in range II, separate phases of alpha- and/or beta-Ca3(PO4)2 and a modified phase of apatite, may increase or decrease the solubility depending on the Ca/P ratio and the resultant amounts of alpha-, beta-Ca3(PO4)2 formed. Modifications in tooth enamel apatite effected in range I are expected to decrease its solubility; the formation of pyrophosphate in this range may have a substantial effect on reducing the solubility rate.(ABSTRACT TRUNCATED AT 400 WORDS)

Biological Transport, Active↗

The extraction of phosphoproteins from bovine dentin.

The phosphoprotein obtained by the neutral pH tris buffer extraction of acetic acid demineralized bovine dentin has been compared with the phosphoprotein extracted directly during the neutral pH EDTA demineralization process. The phosphoproteins isolated by DEAE-cellulose chromatography from the neutral pH EDTA demineralization extract are not identical to those isolated by the same procedure from the dentin which had been subjected to acid demineralization. The two demineralization procedures yield phosphoproteins different in amino acid content and in presence of 260 nm UV absorbing moiety. Even after sequential acid demineralization, trisbuffer extraction and EDTA extraction, the residual dentin contains phosphoprotein. A peptide fragment containing both collagen and phosphoprotein moieties has been isolated following digestion and cleavage of the insoluble dentin collagen with cyanogen bromide. The acid demineralization process appears to be accompanied by degradation which removes both protein and non-protein components from the phosphoprotein.

Acetates↗