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Biomedical subjects

E C Moreno

Publications and source records attributed to E C Moreno.

At least 55 records · Page 3Linked to original sources
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↗
PubMed1986

Saturation of human salivary secretions with respect to calcite and inhibition of calcium carbonate precipitation by salivary constituents.

The state of saturation of human salivary secretions with respect to calcite has been investigated. This property cannot be calculated exactly because of uncertainties in the values of the solubility product constant of calcite, the dissociation constants of carbonic acid, and PCO2 values of saliva. Minimum and maximum limits for this saturation, however, can be established using appropriate values for the constants and salivary PCO2. Values that give the minimum degree of saturation show that 8 of the 70 samples of human saliva investigated would be supersaturated with respect to calcite, while 64 of the 70 samples appeared to be supersaturated when values giving the maximum degree of saturation were used. In the latter case, the ratio of ionic activity products to solubility product was above 10 for several samples and over 18 for the most supersaturated sample. Since these results show that supersaturation of saliva with respect to calcite may be a common condition, human salivary secretions were investigated for the presence of inhibitors of calcite precipitation. Inorganic phosphate and the acidic proline-rich proteins, known to be inhibitors of calcite precipitation, and human salivary statherin, now shown to have a similar activity, are present in saliva at concentrations considerably higher than those required to inhibit calcite precipitation under salivary conditions. Quantitatively, phosphate is by far the most important inhibitor of calcite precipitation present in saliva, suggesting that inhibition of calcite precipitation by the macromolecules may be of secondary significance. It seems more likely that the function of these molecules is to inhibit precipitation of calcium phosphate salts, as previously proposed. These different inhibitory activities, however, are likely to be factors in the differences in composition of oral and dental calculi in different species, and may need to be considered in the formation of calcite stones in the pancreas.

Calcium Carbonate↗
PubMed1986

Effect of low levels of fluoride in solution on enamel demineralization in vitro.

The effect of low levels of fluoride in solution on in vitro enamel demineralization has been studied. Extracted human teeth were exposed to 0.1 M lactate solutions (at pH 4.3) which were partially saturated with respect to enamel mineral and which contained between 0.004 and 1 ppm fluoride. Enamel demineralization was monitored using SEM and polarized light microscopy. It was observed that, in the absence of fluoride, rapid enamel demineralization occurred, resulting in the formation of cavitations within 72 hours. Using the same demineralizing medium containing as little as 0.024 and 0.054 ppm fluoride, we observed a remarkable protection of the enamel surface. Subsurface enamel demineralization was, however, observed under these conditions as well as in a solution containing 0.154 ppm fluoride. When the demineralizing solution containing 1 ppm fluoride was used, no mineral loss was detected. The observed inhibition of enamel demineralization was also shown to be associated with a significant uptake of fluoride by the enamel mineral. These observations have been noted to correlate with an increase in solution supersaturation with respect to fluoridated apatitic species. The results obtained were found to be consistent with a proposed hypothesis that the net rate of enamel demineralization will be reduced in a demineralizing medium supersaturated with respect to less soluble fluoridated phases, due to the enhancement of the precipitation rate of fluoridated apatitic phases relative to the rate of dissolution of the original enamel surface.

Dental Caries↗
PubMed1985

Evaluation of 2-phosphono-butane 1,2,4 tricarboxylate as a crystal growth inhibitor in vitro and in vivo.

The inhibitory activity of 2-phosphono-butane 1,2,4 tricarboxylic acid (PBTA) in the formation of hydroxyapatite (HA) was studied in vitro and in vivo. PBTA, at 4 ppm, inhibited the spontaneous formation of HA in vitro from a supersaturated solution. PBTA, at 2 ppm, completely inhibited the crystal growth of HA; at lower concentrations, a direct relationship was found between the reduction of the initial precipitation rates and PBTA concentrations in the solution. The effects of PBTA on human dental enamel in vitro were also evaluated at pH 5.0 and 7.5. Equimolar levels of EDTA served as the controls. In comparison with EDTA, the dissolution of enamel induced by PBTA was negligible. One percent solution of PBTA was evaluated in a rat calculus assay. In comparison with a placebo solution, it significantly (p = 0.05) reduced calculus formation when applied topically. It was also tested against calculus formation in beagle dogs. A topical application once a day of a 1% solution at pH 7.0 reduced calculus formation by 84% for 16 weeks. Analysis of these data suggests that the agent effectively reduces calculus formation in vivo.

Animals↗
PubMed1985

Contribution of stereochemical interactions in the adhesion of Streptococcus sanguis C5 to experimental pellicles.

Parameters for the adsorption of Streptococcus sanguis C5 to experimental salivary pellicles were determined to assess the relative contributions of stereochemical and less specific hydrophobic-electrostatic interactions in this process. S. sanguis C5 cells possess hydrophobic surfaces and also an adhesin which exhibits stereochemical binding to sialic acid-containing salivary receptors. The adsorption of S. sanguis C5 to untreated pellicles was significantly better described by a two-site adsorption model than by a one-site model (p less than 0.001). One class of binding sites had an affinity for the organism that was more than 400 times that of the second class of binding sites. However, adsorption of the organism was better described by a one-site model to asialo pellicles prepared by neuraminidase treatment which destroyed the receptors for the organism's adhesin. The affinity of the organism to sites in asialo pellicles was low, and approximated that to the low-affinity sites in untreated pellicles. These observations suggest that, in the absence of stereochemical interactions involving the organism's adhesin and specific salivary receptors, S. sanguis C5 cells only exhibit low-affinity binding to experimental pellicles. This was substantiated by studies which showed that adsorption of the organism to albumin-coated hydroxyapatite and to polystyrene beads was also better described by a one-site model, and it occurred with only low affinity.

Adhesiveness↗
PubMed1985

Importance of high pKA acids in cariogenic potential of plaque.

Through the use of computer simulation, it is shown that anions of high pKA acids (e.g., acetic, propionic, butyric), present in resting plaque fluid, act as effective buffers during the production of stronger acids (e.g., lactic) by oral micro-organisms. Using reported organic acid compositions for plaque, obtained from caries-resistant and caries-susceptible individuals, analyzed at various times following sucrose exposure, it is shown that the calculated pH values of the aqueous phase of plaque describe typical Stephan curves. Furthermore, it is shown that for higher acid anion contents in resting plaque, a higher degree of saturation of the aqueous plaque phase with respect to enamel is maintained during acid production, resulting in a lower demineralization potential of plaque. Enamel demineralization experiments conducted in vitro confirmed the significance of the results of the computer simulations, thus providing evidence for a correlation between the known metabolic activity of plaque (organic acid composition), physical chemistry of plaque, and caries susceptibility.

Acetates↗
PubMed1985

Micro-analytical determination of pH, calcium, and phosphate in plaque fluid.

Micro-analytical techniques for the determination of calcium, phosphate, and pH in a small volume (less than 0.25 microliter) of plaque fluid are described and evaluated. The accuracy and the precision of the techniques were compared with those for standard macrotechniques applied to a large pooled plaque fluid sample. The results obtained for the micro-analysis of pooled plaque fluid were in excellent agreement with those obtained by macromethods. The described techniques were also used to analyze plaque fluid obtained from single quadrants of the oral cavities of five individuals. In this fashion, it was determined that, although a significant variation in plaque fluid composition exists between the quadrants, a greater variation exists between subjects. Analyses of plaque fluid obtained from six individuals, following sucrose exposure, were also conducted. The pH value of the fluid changed with time, following a typical Stephan curve, with a minimum value occurring between 15 and 30 minutes; following this, the pH increased to a value near that for resting plaque. An inverse relationship between pH and calcium and phosphorus concentrations was observed. It is noted that the described techniques are sensitive enough to carry out the above analyses both accurately and precisely using plaque obtained from a single quadrant.

Adult↗
PubMed1984

Adsorption of molecules of biological interest onto hydroxyapatite.

Equilibrium and kinetic experiments were conducted to investigate the factors determining the adsorption of salivary macromolecules onto hydroxyapatite. Using amino acids and other small adsorbates, it was determined that the carboxyl attached to the alpha carbon does not appear to adsorb onto HA and the affinities of side-chain carboxyls are much smaller than that of the phosphate group (phosphoserine). Hydroxyl (serine) displays an extremely high affinity, but its adsorption site on HA is different and the number of such sites is much smaller than found for the rest of the functional groups investigated. It is shown that the information obtained from small molecules cannot be readily applied to prediction of the adsorption behavior of salivary macromolecules and polypeptides. The kinetics of adsorption of the salivary phosphopeptide statherin, a polyaspartate, and the salivary prolinerich phosphoprotein PRP3 are consistent with the reversibility of the adsorption process; no conclusion was possible in the case of the protein PRP1. Apparent irreversibility cannot be explained on the basis of multipoint binding or the properties of the carboxyl versus phosphate group; it appears that secondary structure determines to a significant extent the adsorption properties of the macromolecules. Calculation of the thermodynamic molar quantities of adsorption of PRP1, PRP3, and L-ASP onto HA showed that the process is entropically driven. The functional relationship between partial molar entropy and adsorption coverage is similar for the two proteins, but quite different from that for aspartate. Explanations for these results are advanced on the bases of changes in structure configurations and displacement of water from the adsorbate and the adsorbent surface, the second factor being the dominant one in the adsorption of a small molecule such as L-ASP.

Absorption↗
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

Relationship between concentration of human salivary statherin and inhibition of calcium phosphate precipitation in stimulated human parotid saliva.

Human salivary secretions are supersaturated with respect to the calcium phosphate salts which form dental enamel, a property which provides important protection for the teeth. We previously proposed that statherin, a 43-residue phosphopeptide, plays a key role in this protective system by inhibiting or delaying potentially harmful precipitation of calcium phosphate salts in the salivary glands and mouth. The purpose of the present study was to determine if the concentrations of statherin in saliva, despite their wide normal range, are high enough to fulfill this function. Concentrations of statherin in stimulated human parotid saliva samples from 36 female and 32 male subjects, aged from 17 to 30 years, were determined by a single radial immunodiffusion method. Values found ranged from 3.0 to greater than 27.3 microM, with a mean value of 12.8 (S.D. +/- 5.46) microM. At concentrations below these values, statherin inhibited spontaneous precipitation of calcium phosphate salts from an assay system which was more supersaturated with respect to dicalcium phosphate dihydrate, and comparably supersaturated with respect to hydroxyapatite, than were human saliva samples. The inhibitory activities of five of the 65 stimulated parotid saliva samples assayed were greater than would be anticipated from their statherin concentrations. This unexplained discrepancy is not associated with the presence of the acidic proline-rich proteins in saliva, although these proteins also affect calcium phosphate precipitation. The results of this study show that statherin is present in stimulated human parotid saliva at concentrations and levels of activity which are consistent with its proposed biological function, and support the proposal that statherin plays a significant role in a system which provides a protective and reparative but stable environment for the teeth.

Adolescent↗
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↗
PubMed1983

Effects of EDITEMPA on dental calculus and caries formation in vivo.

N,N,N',N'-ethylene diamine tetra (methylene phosphonic acid)--EDITEMPA--was highly effective in reducing dental calculus formation in beagle dogs when applied topically as a 1% solution or in a mouth rinse. A dose-response study in 40 beagles for 12 weeks with dentifrices containing 1, 2, 2.75, and 2.75% EDITEMPA + 0.1% NaF gave a significant (alpha = 0.05) reduction in calculus formation. Based on these results, it was also concluded that 0.1% NaF did not affect the anticalculus activity of 2.75% EDITEMPA. The effect of EDITEMPA dentifrice on dental caries in rats was also studied. A dentifrice containing 1.69% EDITEMPA significantly (alpha less than 0.01) reduced smooth and fissure caries in rats when applied topically. Collectively, the data suggested that EDITEMPA was an effective anticalculus agent and that it did not damage developing enamel in vivo.

Animals↗