Effects of salivary film velocity on pH changes in an artificial plaque containing Streptococcus oralis, after exposure to sucrose

Results from a computer model suggest that following exposure of dental plaque to sucrose, the rate of clearance of acids from plaque into the overlying salivary film will be greatly retarded at low film velocities. This was investigated with an in vitro technique in which artificial plaque containing S. oralis cells was exposed to 10% sucrose for one min. The pH at the proximal (P) and distal (D) undersurfaces of the plaque (0.5 or 1.5 mm thick) was then monitored during the passage of a 0.1-mm-thick film of a sucrose-free solution over the surface. Over the range of salivary film velocities that have been estimated to occur in vivo (0.8-8 mm/min), lower minimum pH values and increased times for the pH to recover toward neutrality occurred at the lower salivary film velocity. Lower pH values were also reached with the 0.5- than with the 1.5-mm-thick plaque. P/D pH gradients, with a lower pH distally, developed at film velocities of 0.8 and 8 mm/min, and the gradients were much more pronounced at the lower velocity. No P/D pH gradients developed when the film velocity was 86.2 mm/min. Incorporation of dead S. oralis cells into the plaque at percentages up to 57% reduced the extent of the pH fall and prolonged the recovery of the pH toward neutrality. The results support the prediction that, other factors being equal, plaque located in regions of the mouth with low salivary film velocity will achieve pH values lower than those of plaque of identical dimensions and microbial composition located in areas where salivary film velocity is high.     

Urea concentration in minor mucous gland secretions and the effect of salivary film velocity on urea metabolism by Streptococcus vestibularis in an artificial plaque

Our purpose was to determine the urea concentration in minor mucous gland (MMG) secretions and the pH at proximal and distal aspects of the lower surface of artificial plaque in vitro during infusion of urea solutions over the surface, at different film velocities. Saliva is present in the mouth as a slowly moving film (ca. 0.1 mm thick) with an estimated velocity in the range of 0.8-8.0 mm/min. At low velocities, due to the accumulation of bacterial products, a progressive increase in their concentration may occur in both the plaque and the overlying salivary film at the distal edge (where the film leaves the plaque). S. vestibularis, an oral micro-organism possessing ureolytic activity, was combined with 1% agarose, to give a urease Vmax similar to that of natural plaque. The artificial plaque was in the chamber (6.0 x 6.0 square and 0.5 or 1.5 mm deep) of a diffusion apparatus, and a urea-containing artificial saliva (3.3 or 13.2 mmol/l) was infused over the surface, as a film 0.1 mm deep, at velocities of 0.8, 8.2 and 86.2 mm/min. At the lower (physiologically normal) urea concentration and the two lower film velocities, most urea appeared to be metabolized at the proximal end of the plaque, which developed a higher pH. At the higher urea concentration, and a film velocity of 8 mm/min, a higher pH was found at the distal end. This was probably due to the combination of greater urea availability and a reduced rate of ammonia loss distally. At a film velocity of 86.2 mm/min, proximal/distal pH gradients did not develop.(ABSTRACT TRUNCATED AT 250 WORDS)     

pH changes of mixed biofilms of Streptococcus mutans and Candida albicans after exposure to sucrose solutions in vitro

ObjectiveThis study aimed to standardize an in vitro experimental model able to reproduce the pH changes that occur in dental biofilm under in vivo conditions, using a mixed biofilm of Streptococcus mutans and Candida albicans.DesignBiofilms were developed for 96 h, and exposed to three different concentrations of sucrose (10, 20 or 30%) during 1, 3 or 5 min. The pH was measured before exposure to sucrose, immediately after its removal from the biofilms, and at 1, 3, 5 and 10 min after removal.ResultsSucrose solutions at 10 and 20% required 1 min to significantly reduce the biofilm pH, while for 30% sucrose a significant reduction was already seen immediately after its removal, even for the shortest exposure time. For an exposure of 3 min to 20% sucrose, the biofilm pH attained the critical value for hydroxyapatite dissolution when measured 1 min after sucrose removal, followed by a recovery phase.ConclusionsA mixed biofilm of S. mutans and C. albicans exposed to a 20% sucrose solution for 3 min exhibited a pattern of pH change similar to that observed in vivo, despite at a higher speed when compared to in vivo conditions.     

The effects of different concentrations of sucrose, fructose, and glucose on pH changes by Streptococcus mitior in an artificial mouth

The aim of this study was to determine the effects of the initial sucrose (S) concentration, as well as those of fructose (F), glucose (G), and invert sugar (F/G), on the pH developed by a layer of S. mitior, to represent dental plaque, in an artificial mouth which simulates the process of oral sugar clearance. At S, F. G, and F/G concentrations of 2% and with normal oral sugar clearance rates, the bacteria produced a smaller pH fall from S than from the other sugars; at initial concentrations of 20%, however, the differences were not significant. With constant S concentrations of 0.5-35%, the minimum pH reached was 4.03 +/- 0.14 (S.D.); with S concentrations of 50% and above, slightly but significantly higher values (4.41 +/- 0.34) occurred. However, with normal sugar clearance, the pH fall was much less than with a constant concentration and was dependent on the S concentration over the range of 0-10%, but was independent at higher concentrations. Exposure of the bacteria to S for as short a period as two min during normal sugar clearance gave a nearly maximum pH fall. This suggests that rinsing the mouth with water more than two min after consumption of sucrose in liquid form will have very little effect in reducing the pH fall in dental plaque. A more appropriate method for reducing acid formation by dental plaque would be consumption of a salivary stimulant which would increase the flow rate and buffer capacity of the saliva.     

Effect of chewing gum containing sodium bicarbonate on human interproximal plaque pH

The effect of chewing sorbitol gum containing sodium bicarbonate on interproximal plaque pH was determined by use of an ion-sensitive field-effect transistor (ISFET) electrode system with four human volunteers. Plaque pH was lowered by 2 min of chewing of a sucrose-containing toffee. After 20 min, either sodium bicarbonate or control gum was chewed for 10 min, and the pH was followed for an additional 20 min. The minimum pH after toffee chewing was elevated by both the control gum (4.5 +/- 0.3 to 5.2 +/- 0.5) and the sodium-bicarbonate-containing product (4.3 +/- 0.3 to 6.1 +/- 0.6) to levels which were significantly different (p less than 0.01). The rate of rise in pH was 2.6 times faster with the gum containing buffer compared with the control gum. In comparison with the control gum, the sodium bicarbonate gum caused the pH to remain at a higher level, approximately 0.5 pH units, for 20 min after gum chewing. The addition of sodium bicarbonate to gum containing sorbitol markedly enhanced its capacity to cause and maintain an elevation of interproximal plaque pH previously lowered by exposure to fermentable carbohydrate. Sodium bicarbonate may be useful in products designed to reduce the acidogenic challenge to the teeth following food ingestion.     

Effect of sorbitol gum chewing on plaque pH response after ingesting snacks containing predominantly sucrose or starch

The purpose of this study was to determine the effect of chewing a sorbitol gum (Trident) for 10 minutes on interproximal plaque pH changes following ingestion of selected sucrose- or starch-containing foods. The snacks containing predominantly sucrose (and/or simple sugars) were chocolate bar, cream-filled cupcakes, cream-filled sandwich cookie, cherry pie and raisins. The snacks containing predominantly starch were oat cereal, granola bars, pretzels, potato chips and corn chips. Plaque pH responses were monitored using an indwelling wire-telemetry system in five adult panelists. The test design involved two sets of 5 x 5 Latin square randomization in which each set consisted of two series of tests. In the first series of tests, the fasted, resting plaque pH was recorded for 5 minutes, panelists ingested the designated snacks for 2 minutes, and the pH response was monitored for the remainder of a 2-hour period. In the second series of tests, the same procedure was followed through the snack ingestion. After the pH response to the snack was monitored for 15 minutes, the panelists were asked to chew one stick of sorbitol gum for 10 minutes and the pH response was then monitored for the balance of the 2-hour period. Results indicated that both the sugar- and starch-containing snacks tested in this study caused significant decreases in interproximal plaque pH. Chewing a sorbitol gum after ingestion of the snacks significantly reduced the demineralizing potential of the plaque. The chewing of sorbitol gum following the ingestion of snacks can be recommended as an adjunct to other caries-preventive oral hygiene measures.     

Salivary clearance of sugar and its effects on pH changes by Streptococcus mitior in an artificial mouth

We recently developed a computer model of oral sugar clearance (Dawes, 1983), and an artificial mouth has now been constructed which will allow variation in the salivary parameters identified in the model and the study of their effects on bacterial acid production. Using a thin layer of S. mitior, strain 572, over a miniature Sb electrode to measure bacterial pH changes during sugar clearance from the "mouth", we found that three consecutive sets of "Stephan curves" could be obtained with the same bacteria during one day. In each of several experimental series, one salivary parameter was varied, while other parameters were held constant. The maximum pH decrease and the surface area of the registered Stephan curve (delta pH.min) were used as measures of acid production. The results indicate that the volume of "saliva" in the mouth before and after swallowing, the unstimulated salivary flow rate, and the buffer capacity had significant influences on the extent of the pH changes. Other factors, such as the maximum flow rate, the delay between start of stimulation and maximum flow, the volume of saliva in the mouth at time zero, and the taste threshold for sugar, were of lesser importance, confirming some predictions from the computer model.     

Human dental plaque pH, and the organic acid and free amino acid profiles in plaque fluid, after sucrose rinsing

The relationship between these factors was studied in plaque and plaque fluid samples taken at intervals during the Stephan pH curve following a sucrose mouth rinse. Levels of lactate rose after the rinse, then fell during the pH recovery phase. Levels of acetate, propionate and phosphate fell after rinsing, then rose again. Amino acid concentrations also changed, with many showing a fall followed by a rise; others rising then falling; and some showing a more variable or complex pattern. In resting plaque fluid, only alanine, proline, glutamic acid, glycine and ammonia were present at concentrations above 1 mmol/l. Delta-aminovaleric acid was detected at levels below those that have been found in monkeys. Hydroxyproline and hydroxylysine were consistently detected, levels of arginine were generally low, and those of cystine consistently very low. The results may provide a basis for understanding the complex metabolic interrelations that occur in the course of the Stephan curve and which may reflect or produce the observed pH changes. They suggest that besides the amount of acid produced, the type of acid, buffering power and base production should be considered as determinants of plaque pH.     

咀嚼无糖口香糖对含漱蔗糖溶液后牙菌斑原位pH值的影响

目的 通过对牙菌斑原位pH值变化的动态监测，观察咀嚼无糖口香糖对牙菌斑原位pH值的影响。方法 采用受试者自身对照的临床试验方法，选择16名健康成人志愿者为受试者，年龄23～32岁，其中男性6名，女性10名。首先测定受试者48h菌斑的静止pH值，以及受试者用10％蔗糖溶液含漱1min后在5、10、20和30min时菌斑的pH值，取得受试者的Stephan曲线作为基线对照；而后观察咀嚼两种益达无糖口香糖对含漱10％蔗糖溶液后菌斑pH值变化的影响。菌斑原位pH值的测定采用pH微电极接触法在口内直接测量。结果 含漱10％蔗糖溶液后立即开始咀嚼无糖口香糖可使菌斑pH值在各检测时间点(含漱10％蔗糖溶液后5、10、20和30min)均维持在静止pH水平，无明显下降；含漱10％蔗糖溶液后在5min时开始咀嚼无糖口香糖则使菌斑pH值从含漱蔗糖溶液后5min时的5．59迅速回升至10min时的6．98。结论 受到蔗糖攻击后，咀嚼无糖口香糖可迅速缓冲菌斑的酸性产物，升高菌斑pH值。     

The effect of swallowing frequency on oral sugar clearance and pH changes by Streptococcus mitior in vivo after sucrose ingestion

A theoretical study of oral sugar clearance (Caries Res 17:321-334, 1983) suggested that the unstimulated salivary flow rate (UNSTFR) and the volume of saliva present in the mouth before swallowing (VMAX) can greatly affect the rate of sugar clearance. The object of this study was to determine whether variations in UNSTFR, by water infusion into the mouth, and in VMAX, which can be altered by varying the swallowing frequency, would influence sugar clearance and whether changes in VMAX would affect the extent of the pH fall produced by S. mitior in vivo after sucrose consumption. In three experiments on each of ten adult subjects, UNSTFR was measured initially (mean value = 0.55 ml/min), and a swallowing schedule was calculated so that the volumes swallowed were either 0.3 ml or 1.0 ml. After a 10-second rinse with 20 ml of a 10% sucrose solution, the subjects maintained a swallowing frequency such that the volumes swallowed were either 0.3 ml or 1.0 ml at normal UNSTFR or were 0.3 ml when UNSTFR was increased by infusing water at 0.5 ml/min. In all subjects, clearance of sugar was faster at the higher swallowing frequency and at the higher flow rate, as predicted by the theoretical model. Small acrylic splints were made to fit over the lower incisors of ten subjects and to hold in the floor of the mouth an antimony microelectrode covered by a 0.5-mm layer of S. mitior, which was held in place by a dialysis membrane.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of an essential oil mouthrinse, with and without fluoride, on plaque metabolic acid production and pH after a sucrose challenge

This clinical study evaluated the effect of rinsing with an essential oil-containing antiseptic mouthrinse, with or without 100 mg/kg fluoride ion, on the plaque metabolic acid production and plaque pH response after a sucrose challenge. This observer-blind, randomized study used a three-way crossover design. Twenty-four subjects rinsed with 20 ml of one of the following rinses: (1) essential oil (EO) mouthrinse, (2) essential oil mouthrinse plus 100 mg/kg fluoride, or (3) negative control, for 30 s, twice daily for 16 days. On day 17, 1 h after the last mouthrinse, subjects rinsed with 20 ml of mass fraction 10% sucrose solution for 1 min. Seven minutes after the sucrose challenge, supragingival plaque was collected from molar and premolar teeth. Plaque pH and metabolic acid ions were analyzed using a micro pH electrode and capillary electrophoresis, respectively. The results showed that after EO mouthrinse dental plaque produced 36% less lactate, 36% less acetate and 44% less propionate than after the negative control rinse. The dental plaque also exhibited a pH 0.42 unit higher after EO rinse than after the negative control rinse. These results were not affected by the addition of 100 mg/kg fluoride to the EO mouthrinse. From these results we concluded that this EO antiseptic mouthrinse, with or without fluoride ion, is effective in reduction of plaque acidogenicity after a sucrose challenge.     

A theoretical analysis of the effects of plaque thickness and initial salivary sucrose concentration on diffusion of sucrose into dental plaque and its conversion to acid during salivary clearance

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)     

Effect of an essential oil-containing antiseptic mouthrinse on plaque and salivary Streptococcus mutans levels

BACKGROUND: Clinical studies in which antimicrobial mouthrinses were shown to have significant antiplaque activity most frequently have used gingivitis as the clinically relevant endpoint. However, there is evidence to suggest that mouthrinses containing active agents effective against Streptococcus mutans, such as chlorhexidine, may also have a role in inhibiting dental caries. This clinical study was conducted to determine the effect of 2x-daily rinsing with an essential oil-containing antiseptic mouthrinse (Listerine Antiseptic) on levels of recoverable S. mutans and total streptococci in supragingival interproximal plaque and in saliva. Additionally, a follow-up in vitro study is reported which determined whether a differential susceptibility to the antiseptic mouthrinse exists among different strains of streptococci. METHOD: Following baseline saliva and plaque sampling for quantification of recoverable S. mutans and total streptococci, 29 qualifying subjects were randomly assigned either the essential oil mouthrinse or a sterile water control. They rinsed with 20 ml for 30s 2 x daily for 11 days and once on the 12th day, in addition to their usual oral hygiene procedures. On day 12, saliva and plaque samples were again collected and microbiological quantification performed. The procedures were repeated with the alternate rinse after a 1-week washout period. RESULTS: The essential oil mouthrinse produced respective reductions of 69.9% and 75.4% in total recoverable streptococci and in S. mutans in plaque, and corresponding reductions of 50.8% and 39.2% in saliva. The in vitro study revealed that streptococci from the mutans group were more susceptible to the bactericidal activity of the essential oil mouthrinse than streptococci from the mitis group. CONCLUSIONS: As antimicrobial mouthrinses are most frequently recommended to patients whose mechanical oral hygiene procedures are not adequate for the control of supragingival plaque and gingivitis, this study provides an additional rationale for the inclusion of the essential-oil mouthrinse as an adjunct to daily oral hygiene procedures.     

Salivary concentrations of urea released from a chewing gum containing urea and how these affect the urea content of gel-stabilized plaques and their pH after exposure to sucrose

The objectives were to: (1) determine the salivary concentrations of urea during 20 min chewing of a sugar-free gum containing 30 mg of urea; (2) measure the degree to which this urea would diffuse into a gel-stabilized plaque; (3) study the effect of the urea on the fall and subsequent rise in pH (Stephan curve) on exposure to 10% sucrose for 1 min; (4) model the measurements 2 and 3 mathematically. In point 1, the salivary urea concentration of the 12 subjects peaked at 47 mmol/l in the first 2 min of gum chewing, falling within 15 min to the unstimulated salivary concentration of 3.4 mmol/l. Recovery of urea from the saliva averaged 81.5%. 'Plaques' of 1% agarose or 67% dead bacteria in agarose accumulated urea from the saliva roughly as expected, whereas those plaques containing 8% live and 59% dead Streptococcus vestibularis showed negligible accumulation. Computer modelling showed this difference to be due to urease of live bacteria breaking down the urea as rapidly as it entered the plaque. Simulation of the effect of gum chewing subsequent to initiation of a Stephan curve in the latter type of plaque showed a rapid rise in pH but then a fall again on return to unstimulated conditions. This fall had not been seen in previous studies, with Streptococcus oralis, nor was it predicted by the computer modelling. Neither experimental simulation nor computer modelling suggested that chewing urea-containing gum before exposure to sucrose would have any effect on a subsequent Stephan curve. Thus chewing gum is only likely to inhibit caries when it is chewed after consumption of fermentable carbohydrate, rather than before.     

Streptococcus mutans in plaque after mouth-rinsing with buffers of varying pH value

After a period of mouthrinsing with a phosphoric buffer solution with low pH the proportional distribution of Streptococcus mutans in plaque from occlusal retention sites was significantly higher than after a control period. This indicates that colonization with S. mutans may be favored or that other microorganisms competing with S. mutans are inhibited under these circumstances.     

Streptococcus mutans in plaque after mouth-rinsing with buffers of varying pH value

Abstract – After a period of mouthrinsing with a phosphoric buffer solution with low pH the proportional distribution of Streptococcus mutans in plaque from occlusal retention sites was significantly higher than after a control period. This indicates that colonization with S. mutans may be favored or that other microorganisms competing with S. mutans are inhibited under these circumstances.