Previously, we studied the clearance rates of KCl from agarose gels positioned at different locations in the mouth, and showed that the rates were much slower than when clearance was into a well-stirred solution. We designed the present in vitro study to test the effect on KCl clearance of the velocity of a 0.1-mm-thick film of water flowing over an agarose gel of the same diameter and composition as those used in vivo. The thickness of the salivary film overlying dental plaque has been estimated to be about 0.1 mm, and we assumed that when clearance rates in vitro matched those found in vivo, velocities of the fluid film (in vitro) and the salivary film (in vivo) must be equal. On this basis, it was calculated in the present experiments that when salivary flow was unstimulated, the velocity of the salivary film at the level of the teeth varied between about 0.8 mm/min (upper-anterior buccal region) and 8.0 mm/min (lower-anterior lingual region). When salivary flow was stimulated, this was estimated to increase the velocity of the salivary film from 2 to 40 times, depending on the location in the mouth. It is postulated that the slow movement of the salivary film when flow is unstimulated allows for accumulation of diffusants from dental plaque, which reduces the concentration gradient for diffusion from plaque and prolongs the clearance time of such metabolic products as acid.

Cultures of Streptococcus mutans MFe28 (serotype h) were grown with differing extracellular polysaccharide (EPS) content. Biochemical and physicochemical characteristics considered relevant to caries were measured. Acid production parameters measured in a pH-stat were: Vm = 0.76 +/- 0.14 mumol/g/sec (wet weight); apparent Km (acid production) = 100 mumol/L; molar yield = 1.97 +/- 0.25 mol acid/mol glucose. Acid anion inhibition of acid production was also noted. Buffering by the pure washed bacterial residue required approx. 112 mumol of base/g (wet weight) of residue to change the pH from 4 to 6.5, and this dropped almost to zero as the EPS content increased to 100%. Diffusion coefficients (D) in the residues were independent of EPS content over a wide range. When the effusion method was used, De (glucose) and De (acetate) were (3.26 +/- 0.6) and (5.05 +/- 0.8) x 10(-6) cm2/sec, respectively. The extracellular fluid fraction, measured by inulin exclusion, increased from 0.33 for the pure bacteria to 0.78 for the pure EPS. It is shown how, by these factors alone, and without any need for diffusion restriction, plaque EPS may lead to a lower pH at the tooth surface, thus increasing the cariogenic challenge.

By means of micro-equilibrium dialysis, calcium binding capacities and affinities were measured in three different oral bacteria, and the effects of extracellular polysaccharide, pH, and aggregation were investigated. Binding capacities of 31.0 +/- 2.1 (C. matruchotii), 34.7 +/- 3.7 (S. sanguis), and 41.5 +/- 5.4 (S. downei) mumol calcium/g wet weight of cells were found at pH 7.0, falling to 21.4 +/- 0.8 mumol calcium/g wet wt. cells at pH 5.0 for S. downei. Dissociation constants were found to vary between 0.78 +/- 0.24 and 1.77 +/- 0.30 mmol/L (at pH 7.0, depending on species), and between 0.62 +/- 0.04 and 1.77 +/- 0.30 (in the pH range 5.0 to 7.0, for S. downei only). Examination suggested that at pH 7.0 calcium-facilitated bacterial association occurs in the streptococci with calcium uptake curves analogous with those of positively cooperative systems. Desorption of calcium from aggregated S. downei suggested that the mechanism of desorption differed from that of uptake. This may be an important factor in plaque formation and in the binding of cells to the surface of formed plaque. Plaque calcium forms a reservoir, readily released by a pH drop, which may increase plaque fluid saturation and reduce demineralization.

A mathematical model, written in FORTRAN, has been developed to simulate the interrelated processes of salivary sucrose clearance from the mouth, diffusion of sucrose into dental plaque, and conversion of sucrose to acid and glucan. Reaction of acid with enamel is not included in the model. A total of 28 parameters can be varied by the user, and the relative importance of the different factors affecting acid formation can be assessed. The output of the program gives sucrose and acid concentrations and pH at different depths within the plaque. The initial variables studied were plaque thickness, the salivary sucrose concentration, and the duration of exposure of the plaque to sucrose. Stephan curves typical of those recorded in vivo were generated by the model. With any particular salivary sucrose concentration, there was an optimum plaque thickness at which a minimum pH was achieved at the enamel surface, with very thin or thick plaque samples producing a smaller pH fall. With thick plaque, the minimum pH was often not achieved at the inner surface but at some intermediate depth, which may explain the location of early caries lesions in fissures. The extent of the pH fall at the inner surface and the duration of the pH-minimum region of the Stephan curve were directly related to the initial salivary sucrose concentration and to the duration of exposure to sucrose prior to normal salivary clearance. Simulation of a water rinse at as short a time as two min after the beginning of normal salivary sugar clearance showed that this procedure had only a very small effect on the shape of the Stephan curve.(ABSTRACT TRUNCATED AT 250 WORDS)

A semi-micro method was used for investigation of the buffering properties of whole plaque, plaque fluid, and washed plaque bacteria. Artifacts associated with titration of samples containing live bacteria were noted and their effects estimated. All three sample types showed minimal buffering in the region of neutrality, with much stronger buffering in the regions pH 4-5.5 and pH 8-9. For the range pH 4-7, almost 90% of the total buffer capacity of plaque appeared to be accounted for by macromolecules of bacterial cell walls and plaque matrix. Extracellular buffers in plaque fluid removable by centrifugation contributed up to 11%. These buffers (probably soluble proteins, peptides, organic acids, and phosphate) are, potentially at least, capable of exchange with saliva. In vitro, bicarbonate (dissolved in the extracellular fluid) contributed only 2-5% of total buffering; there was no evidence of formation of carbamino compounds. However, in vivo, salivary bicarbonate may be important as a continually replenished source of additional buffering.