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T Aoba

Publications and source records attributed to T Aoba.

At least 55 records · Page 3Linked to original sources
PubMed1989

Possible function of matrix proteins in fluoride incorporation into enamel mineral during porcine amelogenesis.

The present study was undertaken to elucidate the mechanism of fluoride incorporation into secretory enamel mineral, with porcine enamel used as a model. Although the fluoride content in the enamel varied greatly among the animals, we observed that the fluoride-to-calcium ratio in the enamel tissue was maximal at the beginning of the secretory stage; the F/Ca ratio decreased (and leveled off) with the advancement of mineralization. In vitro work showed that some of the fluoride in the secretory enamel tissue was removed with the extraction of organic matter, mostly amelogenins. Furthermore, coating hydroxyapatite crystals with enamel matrix proteins resulted in a retardation of fluoride incorporation into the crystals when exposed to fluoride solutions, as a result of an inhibition of apatite reprecipitation. We also confirmed that the growth kinetics of fluoridated apatite onto HA seeds decreased with increasing coverage of the seed surface with the enamel proteins. All the results of the present study strongly suggest that the fluoride incorporation into enamel mineral during the secretory stage may be regulated by the kinetics of mineralization, which is highly dependent on the driving force for precipitation and the presence of proteinaceous inhibitors, mainly amelogenins.

Amelogenesis↗
PubMed1989

Possible roles of partial sequences at N- and C-termini of amelogenin in protein-enamel mineral interaction.

The purpose of this study was to assess the functional significance of homologous sequences of mammalian amelogenins at their N- and C-termini. A porcine 5-kDa fragment corresponding to the N-terminal 45 residues of amelogenins was purified from the secretory enamel. The decapeptide TDKTKREEVD corresponding to the C-terminal 10 residues of amelogenins was synthesized according to conventional solid-phase procedures. The inhibitory activity of both moieties on apatite crystal growth was determined in a supersaturated solution having an ionic composition similar to that of the fluid phase separated from porcine secretory enamel. The 5-kDa amelogenin fragment was sparingly soluble in neutral solutions and (in condensed forms because of aggregation) showed no significant inhibition of crystal growth, whereas the fragment molecules pre-adsorbed onto the seed crystals yielded modest inhibition of hydroxyapatite precipitation. However, their inhibitory activity was significantly lower than that of parent porcine amelogenin (25-kDa molecular mass). The high solubility of synthesized decapeptide allowed us to determine the adsorption isotherm onto hydroxyapatite at 37 degrees C, at an ionic strength similar to that of the enamel fluid. The obtained adsorption isotherm was described by a Langmuir model; the adsorption affinity and the maximum adsorption sites were 6.2 mL/mumol and 0.53 mumol/m2, respectively. As expected from the low adsorption affinity, the peptide showed a much weaker inhibition of apatite crystal growth than the parent amelogenin.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗
PubMed1988

Effect of fluoride in the apatitic lattice on adsorption of enamel proteins onto calcium apatites.

The selective adsorption of enamel proteins onto crystalline calcium apatites having different specific surface areas and various degrees of fluoride substitution was investigated. The proteins were obtained from the outer (close to the ameloblast) layer of secretory enamel of porcine permanent incisors. The adsorption of the enamel proteins was not affected markedly by the variation of specific surface area of the hydroxyapatites used as adsorbents, but it was enhanced substantially with increasing fluoride content in the crystalline lattice. Through the use of SDS- and two-dimensional polyacrylamide gel electrophoresis, it was shown that the originally secreted amelogenin (25 kd) as well as 60-90-kd and 5-6-kd molecules adsorbed most selectively onto the hydroxyapatites and that additional moieties having 21-23-kd and 14-18-kd molecular masses commenced to adsorb onto the apatitic surfaces with increasing degrees of fluoride substitution in the lattice. In contrast, the 20-kd amelogenin, a product partially degraded from the 25-kd amelogenin, showed no significant adsorption, even onto the fluoridated apatites. These results suggest that the retention of proteinaceous matrix in the developing enamel might be affected by the nature of the forming crystals.

Adsorption↗
PubMed1987

Selective adsorption of porcine-amelogenins onto hydroxyapatite and their inhibitory activity on hydroxyapatite growth in supersaturated solutions.

The selective adsorption of amelogenins onto synthetic hydroxyapatite (HA) and their inhibitory activity on the seeded HA crystal growth were investigated using enamel proteins obtained from the outer layer of immature porcine-enamel (soft, cheeselike in consistency) of developing permanent incisors. Special interests were paid to the effect of a postsecretory degradation of the original amelogenin(s) on their adsorption and inhibitory activity. In the adsorption studies, it was apparent that the originally secreted amelogenin (25 K), proline, and histidine-rich protein (2a), as well as the higher molecular weight components (60-90 K), showed a strong adsorption affinity onto the HA. This adsorption of protein 2a was related to its inhibition of the crystal growth of seeded HA in a dilute supersaturated solution. On the other hand, the partially degraded product (20 K) of amelogenins, protein 2b, lost the high adsorption affinity onto the HA, and consequently showed no significant inhibitory activity. The observed selective adsorption of protein 2a onto HA was apparent at pH 6.0 and pH 7.4 even in the presence of dissociative solvents, such as 3 M urea or 2 and 4 M guanidine-HCl; however, this selective behavior was sensitive to changes in pH, and was not displayed at pH values of 7.8 or 10.8. The results suggest that the originally secreted amelogenin 2a may play an active role in amelogenesis, and that enamel mineralization could be regulated by the secretion of amelogenins and their inactivation through partial enzymic degradation, prior to their complete removal.

Adsorption↗
PubMed1987

The enamel fluid in the early secretory stage of porcine amelogenesis: chemical composition and saturation with respect to enamel mineral.

The present study reports on the separation of fluid from soft, "cheeselike" enamel of porcine permanent teeth in the secretory stage, and the determination of its chemical composition. The enamel tissues were dissected from mandibles of 5 to 6-month-old piglets and pooled under mineral oil in centrifuge tubes, and then centrifuged at 1.9-2.4 X 10(5) g for 1-1.5 hours. The yields of the fluid were 44.8 +/- 2.3 (mean +/- standard error) microliter/g of enamel tissue at 1.9 X 10(5) g, and 53.9 +/- 1.9 microliter/g at 2.4 X 10(5) g. A significant finding was that the total Ca concentration of the enamel fluid (3.9-6.0 X 10(-4) M) was lower than that of porcine serum (2.9 X 10(-3) M), reflecting a distinct, compartmentalized microenvironment, isolated from the circulating blood. Another significant finding was that the ionic calcium activity (5.3 X 10(-5) M) in the enamel fluid was one order of magnitude lower than the total Ca concentration. The averaged results of other determinations were: pH, 7.26; total [P], 3.9 mM; [Mg2+], 0.8 mM; [Na+], 140 mM; [K+], 20 mM; [Cl-], 150 mM; [F-], 5 X 10(-3) mM; and osmolality of the fluid, 312 mosmol/kg H2O (in the same range as that of the serum, 310 mosmol/kg H2O). The apparent electrical unbalance of the analytical data, 8.65 meq excess of positive charges, was ascribed to the presence of HCO3- in the fluid; the computed ionic strength was 164 mM.(ABSTRACT TRUNCATED AT 250 WORDS)

Amelogenesis↗
PubMed1987

Proteins in the enamel fluid of immature porcine teeth.

The fluid was separated from the immature soft enamel of porcine permanent teeth in the secretory stage according to procedures reported previously (Aoba and Moreno, 1987). The protein content of the fluid was about 2.8% w/v; its amino-acid composition was characterized by high contents of Pro, Glx, Leu, and His, showing composition similar to that of the 20 kilo-dalton (kd) amelogenin or its C-terminal segments. The two major protein species in the fluid had apparent molecular weights of 13 kd and 11 kd, as determined by SDS electrophoresis; the N-terminal residue of the former was Leu, while that of the latter was Ala. The C-terminal sequence of both of them was -Met-Phe-Ser. By comparison with the published sequence of 20-kd porcine amelogenin, it is concluded that the main fluid constituents were derived by cleavages of N-terminal segments from the 20-kd amelogenin.

Amelogenesis↗
PubMed1984

Inhibition of apatite crystal growth by the amino-terminal segment of human salivary acidic proline-rich proteins.

Inhibition of seeded apatitic crystal growth by human salivary acidic proline-rich phosphoproteins (PRP) has been related to their adsorption onto the apatite seeds. The amino-terminal 30-residue segment of the PRP makes an important contribution to this adsorption. This peptide (PRP1(T1] and its dephosphorylated analogue from PRP3 (PRP3(T1)DP) were prepared. They have identical sequences, except the phosphates at residues 8 and 22 in PRP1(T1) are absent from PRP3(T1)DP. Adsorption of these peptides onto hydroxyapatite and their effect on crystal growth from a defined supersaturated solution was studied. Adsorption behavior was adequately described by the Langmuir adsorption isotherm. The adsorption affinity constant of PRP1(T1) (K = 20,200 ml/mumol) was more than 10 times the corresponding value for PRP3(T1)DP (1,800 ml/mumol), and similar to that of the parent protein, PRP1 (26,200 ml/mumol). Inhibition of crystal growth by the peptides was interpreted in terms of the fractional coverage of the maximum number of adsorption sites (as derived from the adsorption isotherms), suggesting that the molecules block, by adsorption, specific growth sites on these surfaces. Comparison of precipitation kinetics showed that PRP1(T1) is a more effective inhibitor than PRP3(T1)DP at the same initial concentration (10(-6)-10(-7)M). However, on the basis of per mol adsorbed, PRP3(T1)DP displays a greater inhibitory activity; such a behavior is consistent with a more open molecular structure which blocks more growth sites per mol adsorbed than PRP1(T1).(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗
PubMed1984

Preparation of hydroxyapatite crystals and their behavior as seeds for crystal growth.

Samples of crystalline hydroxyapatite [Ca5OH(PO4)3 HA] were prepared by precipitation from aqueous media under a variety of experimental conditions (temperature, concentration of reagents, rates of addition of the reagents, and seeding). The resultant products showed a wide range of particle sizes, i.e., specific surface areas, from 6.39 to 50.1 m2/g. In these preparations, relatively large crystals were obtained with low rates of addition of the reagents or by seeding the precipitating medium. Small differences in super-saturation of the reaction medium can markedly affect the particle sizes and crystalline habits of the resulting products, possibly by altering the processes of nucleation and subsequent crystal growth. When these crystalline materials were used as seeds to study the crystal growth of HA, it was confirmed that the precipitation rate of calcium apatite on seed crystals is highly dependent on the surface areas available for growth, rather than on the particle sizes and amounts of the seed crystals. Small differences in the kinetic runs were observed between the various seed crystals, which can be attributed to differences in the surface properties of these crystals. Additionally, transmission electron microscopy of the seed crystals revealed that some projections form, possibly on the basal planes of the crystals, during crystal growth. Since the growth rate of these projections was greater than the mean growth rate calculated on the basis of changes in solution composition and total surface area, it appears that the kinetics of the growth process is determined, to some extent, by the geometry of the seeds.

Calcium Phosphates↗
PubMed1984

High-resolution electron microscopy of hydroxyapatite grown in dilute solutions.

High-resolution electron microscopy (HREM) was applied to the study of seeded crystal growth of hydroxyapatite (HA) in supersaturated solutions. The HA seed crystals were rod-shaped, elongated along the c-axis, and showed smooth contours. The seed crystals exhibited hexagonal prism facets, and one end was rhombohedrally terminated, whereas the opposite end was blunt. Growth experiments were carried out at 37 degrees C, and solution compositions were carefully selected to avoid the involvement of precursor phases during the HA growth. After five-hour growth, the total amount of precipitation was from 3 to 7% of the initial crystal mass added to the solutions; examination of the crystals by HREM disclosed the formation of projections on the end surfaces of the HA crystals. High-resolution TEM clearly showed lattice fringes with predictable spacings over the entire crystal specimen, including the formed projections. Analysis of selected regions by optical diffraction of the HRTEM fringe negatives showed that the nature and orientation of the projections were similar to those of the underlying seed crystal. High-resolution SEM of the HA crystal after five-hour growth disclosed step-like structures on the prism faces, as well as the projections localized on the ends. These results strongly suggest that more than one growth process may be involved, perhaps related to the distinct faces of the HA seed crystals.

Chemical Precipitation↗
PubMed1982

Crystallinity, solubility, and dissolution rate behavior of fluoridated CO3 apatites.

To study the effect of fluoride on the physicochemical properties of carbonate-containing apatites, two series of fluoridated CO3 apatites with various fluoride contents were synthesized at 80 degrees C and pH 7.4. The alpha-axis dimensions of these apatites decreased with the increase of both fluoride and carbonate contents. The crystallinity of the apatites showed similar patterns of variation with the degree of fluoridation irrespective of carbonate content. These patterns appear analogous to the complex pattern of the fluoridated hydroxyapatites previously reported. In contrast to this unexpected crystallinity phenomena, the apparent solubility of fluoridated CO3 apatites in acetate buffer solutions at pH 4.0 and 6.0 and 37 degrees C decreased monotonically with increasing degree of fluoridation and approached that of fluoridated hydroxyapatites at high fluoride content. Dissolution rate of fluoridated CO3 apatite pellets at pH 4.0 also showed a simple decrease with increasing degree of fluoridation and was extremely sensitive to acid at low fluoride content.

Apatites↗
PubMed1982

ESR and IR studies of carbonate-containing hydroxyapatites.

Synthetic carbonate-containing and carbonate-free hydroxyapatites(CO3-HA and CO3-free HA) have been investigated using ESR and IR absorption, together with X-ray diffraction and chemical analyses. Comparison of the IR spectra between 13C-enriched 13CO3-HA and 12CO3-HA prepared at 100 degrees C and at high pH suggested that the carbonate substitutes for PO4 but not for HPO4. The substitution of CO3 for PO4 in this case resulted in elimination of hydroxyl ions in a hexad axis. The elimination of hydroxyl ions was found to be related to the decrease in crystallinity for the c-axis direction, although so far there has been no direct evidence for the existence of hydroxyl deficiencies. In the case of CO3-free HAs prepared at 80 degrees, 60 degrees, 40 degrees, and 20 degrees C, the decrease in crystallinity accompanied by the decrease in the temperature for preparation was associated with the decrease in crystal size, and no significant elimination of hydroxyl ions would be expected in the absence of the carbonate.

Carbon Isotopes↗