Plaque sampling and telemetry for monitoring acid production on human buccal tooth surfaces

The plaque sampling method and wire telemetry using miniature, glass pH electrodes on the buccal surface of mandibular first permanent molars, compared human dental plaque pH responses to a 10 per cent solution of sucrose. Nine subjects abstained from oral hygiene and were tested in morning sessions on 5 consecutive days. The mean resting plaque pH value for all sessions by plaque sampling was 6.60 +/- 0.18 (mean +/- SD) and by telemetry was 6.56 +/- 0.19. After a 1 min sucrose rinse, 1-day-old plaque showed a decrease in pH approx. 5.5 by both methods. The 2-, 3-, 4- and 5-day-old plaque, the mean minimum pH achieved was 4.69 +/- 0.28 as measured by sampling and 3.94 +/- 0.49 as determined by telemetry. The times taken to reach minimum pH as determined by the two methods were almost identical. The electrode calibration data showed that all the telemetry electrodes responded consistently for all 5 days of study. Scanning electron microscopy and microbiological analysis of electrode tips and enamel replicas revealed that plaque accumulated on glass at the same rate with a similar bacterial composition to plaque formed on clean enamel. It is proposed that glass microelectrodes and wire telemetry are a reasonable means for continuously monitoring plaque pH in situ. Comparison with plaque sampling suggested that telemetric responses reflect the type of plaque which accumulates at particular sites on the dentition.     

Effect of dental plaque age and bacterial composition on the pH of artificial fissures in human volunteers

Changes in sucrose-induced plaque pH profiles and the microbial composition of occlusal tooth surface fissures were analyzed using wire telemetry and bacterial culturing techniques. Four human volunteers wore appliances containing artificial fissures constructed with ion-sensitive field-effect transistor (ISFET) electrodes for 1, 2 and 4 days; 1 subject kept the electrode for 3 weeks. After monitoring the plaque pH response at the base of the fissure to a 10% (w/v) sucrose rinse the plaque was removed and analyzed for total viable bacteria, total and specific streptococci, lactobacilli and Actinomyces spp. One-day-old plaque showed a rapid drop in plaque pH to a minimum of 4.8 +/- 0.2, with 2-day-old plaque showing the most acidogenic pH profile (minimum pH 4.6 +/- 0.2). The 4-day-old plaque response was less acidogenic (minimum pH 5.0 +/- 0.3) than the results from days 1 and 2. Responses from 13- and 21-day-old fissure plaques showed greatly decreased acidogenic responses (day 21 minimum pH 5.7). Viable bacteria recovered from the fissure increased from approximately 4 x 10(6) colony-forming units on day 1 to 1.2 x 10(7) on days 2 and 4 and 1.7 x 10(7) on day 21. Streptococci (greater than 50%) and Actinomyces (greater than 10%) dominated in the fissure plaques and their levels were related to minimum pH. Since fissure plaque of all ages tested contained high concentrations of acidogenic bacteria, the decreased acidogenic response at the base of fissures with increasing plaque age suggests that maturing fissure plaques provide an increasingly greater diffusion barrier to fermentable carbohydrates.     

Comparison of in vivo human dental plaque pH changes within artificial fissures and at interproximal sites

Ion-sensitive field-effect transistor (ISFET) pH electrodes were used to monitor changes in plaque pH at the base of artificial occlusal surface fissures and at interproximal sites. Bovine enamel was used to construct fissures (1.5 x 0.1 x 1.0 mm) containing a small ISFET electrode. The fissures were fixed to carrier appliances and worn by 4 human volunteers. After plaque accumulation for 4 days changes in pH were monitored by wire telemetry following 1-min rinses with 10% solutions of either sorbitol or sucrose. Results were compared to data obtained from interproximal sites in the same subjects. Responses to sorbitol in the fissure and on the proximal surfaces were minimal and showed no significant difference in minimum pH (5.9 +/- 0.4 and 6.1 +/- 0.3, respectively) and area under pH 7.0. The response to sucrose at the two sites was very different revealing unique pH profiles which were statistically significantly different with regard to minimum pH (5.0 +/- 0.3, fissure and 4.3 +/- 0.2, proximal) and area under pH 5.7. Thus, the acidogenic potential of fermentable carbohydrate at two caries-prone sites in the human dentition is significantly different and conclusions based on interproximal telemetry measurements may not be applicable to occlusal surface fissures.     

The acid-tolerant microbiota associated with plaque from initial caries and healthy tooth surfaces

The intent of this study was to compare the inherent acid tolerance of bacteria in samples of dental plaque from tooth sites in subjects with and without initial caries. Plaque was collected from approximal surfaces showing early enamel caries and from healthy tooth surfaces in the same subjects, as well as from enamel surfaces of caries-free individuals. In addition to plating on blood agar, the plaque samples were plated directly on non-selective solid agar medium buffered to pH 7.0, 6.0, 5.5, 5.0, 4.5 and 4.0 to avoid any loss of adaptation to acid during primary isolation of plaque bacteria. The results showed that approximately 50% of the total cultivable plaque microbiota from caries, as well as healthy tooth sites, was able to grow at pH 5.5 and 1% at pH 5.0, pH values regarded as critical for the demineralization of tooth enamel. At pH 5.0, members of the genus Streptococcus were the dominant group, but mutans streptococci accounted for less than half of the streptococcal viable count. The other acid-tolerant streptococcal isolates included Streptococcus anginosus, Streptococcus constellatus, Streptococcus gordinii, Streptococcus intermedius, Streptococcus mitis, Streptococcus oralis, Streptococcus salivarius and SStreptococcus sanguis. Analysis of the results indicated that the mutans streptococci in dental plaque were highly variable with respect to acid tolerance, and that both caries and healthy sites harboured significant numbers of mutans streptococci that were not acid-tolerant.     

Mutans streptococci and lactobacilli in plaque on a leucite-reinforced dental ceramic and on a calcium aluminate cement

In this in vivo study, the proportions of mutans streptococci and lactobacilli in plaque were examined (1) on proximal surfaces of bonded, leucite-reinforced ceramic crowns and (2) on class V restorations of calcium aluminate cement (CAC). The examined proportions were intraindividually compared with those of resin composite and enamel. Mutans streptococci and lactobacilli in samples from plaque that was accumulated for 10 days on the following surfaces were determined by cultivation on blood agar plates and species-selective plates: (1) proximal leucite-reinforced ceramic crown, class II composite and enamel (n=11); and (2) class V restoration of CAC and composite, and enamel (n=17). Mutans streptococci and lactobacilli in the samples were distributed in three groups: 0, >0–1, and >1% of total bacteria. The surfaces with detected mutans streptococci were similarly distributed between the materials and enamel. The highest proportion of mutans streptococci and lactobacilli were observed on ceramic followed by composite and enamel. A higher proportion of lactobacilli, but not of mutans streptococci, was detected on enamel compared to CAC and composite. However, no significant differences were found between the surfaces. Conclusively, the materials investigated did not show different relative proportions of mutans streptococci and lactobacilli in plaque, compared to enamel.     

Enamel microhardness change and plaque pH measurements in an intra-oral model in humans

Changes in plaque pH and microhardness of bovine enamel slabs were evaluated with a seven-day intra-oral cariogenicity test (ICT). The test enamel slabs were mounted in prosthetic appliances with a Dacron mesh cover for enhancement of microbial colonization. Three percent solutions of sucrose, sorbitol, and xylitol were evaluated as four daily extra-oral immersions of 10 min each, for seven days, and the results were compared with baseline experiments (no daily immersions). The pH was measured with antimony electrodes on one-day and seven-day ICT plaque samples that were challenged with a one-minute immersion in the studied substrates. Plaque samples in the baseline experiments were challenged with 3% sucrose. The enamel softening was assessed with measurements of microhardness. Sucrose challenge caused pH depression with both the baseline and the sucrose-immersed plaque. Sorbitol and xylitol challenge did not depress the plaque pH. Compared with the baseline, sucrose immersions caused enamel softening; sorbitol and xylitol did not.     

Comparison of the plaque microflora from natural and appliance-borne enamel surfaces

Human enamel sections and slabs, mounted on a mandibular removable appliance, were worn by 5 adult subjects for a 1-week period. Plaque was allowed to accumulate on the in situ test sites and on the adjacent natural dentition. At the end of the experimental period, the plaque microflora associated with (1) the enamel sections, (2) the enamel slabs, and (3) the acrylic base of the appliance test site was compared with that obtained from lingual and interproximal areas of the lower molar teeth. In addition, the acid anion and pH profiles of plaque obtained from both the exogenous and natural tooth surfaces were also determined. Although some quantitative differences were found between the proportions of isolates obtained from the different enamel surfaces, qualitatively the microflora was very similar, and no significant differences were found in the plaque lactate/acetate ratios or pH measurements following a sucrose mouthrinse. Thus, human tooth specimens mounted on the intra-oral device produced a plaque ecosystem similar to that present on the adjacent natural dentition, suggesting that the model is suitable for studies on early plaque development and the microbiology of enamel demineralization.     

Enhanced gingivitis in the deciduous and permanent dentition An experimental study in the dog

The aim of the experiment was to analyze the reaction of the marginal gingival tissues to 21 days of plaque formation on buccal tooth surfaces in the deciduous and permanent dentition of beagle dogs. In order to enhance the formation of plaque, the buccal surfaces of the experimental teeth were coated with a composite filling material. 5 beagle dogs were used. The animals were monitored during 2 periods, called period A (42 days during the deciduous dentition) and period B (42 days during the permanent dentition). The dogs were 10 weeks old at the initiation of period A. Following 3 weeks of plaque control, a groove was prepared into the enamel of the buccal surfaces of the mandibular right 3rd (03P) and 2nd (02P) premolars. A cotton ligature was subsequently attached to the groove using an enamel/etch-technique and a composite filling material. The groove and the ligature did not interfere with the gingival margin but the composite material extended into the subgingival niche. The plaque control measures were abandoned. The animals formed plaque during the following 21 days. A clinical examination was performed and subgingival bacteria sampled on day 21. Moreover, biopsies were harvested from the 03P and 02P tooth regions. The biopsies were prepared for histometric and morphometric analyses. A 2nd plaque control regimen was initiated. Period B started when the dogs were 15 months old. Following 3 weeks of enhanced plaque control, a cotton ligature was attached as described above at the buccal surfaces of the mandibular left 3rd (P3) and 4th (P4) premolars.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plaque formation in vivo and bacterial attachment in vitro on permanently hydrophobic and hydrophilic surfaces.

Highly hydrated polyethylene oxide (PEO) films represent one type of surface modification which may interfere with biofilm formation. Protein adsorption and saliva-mediated bacterial adherence were investigated in vitro on normal and hydrophobized glass surfaces and on glass surfaces with immobilized PEO films. More protein and bacteria bound to untreated compared to hydrophobized and PEO-treated glass. Pellicle and plaque formation was also studied in vivo on ceramic crown surfaces either untreated, hydrophobized or with immobilized PEO films. Pellicle and plaque formation was similar on the untreated ceramic and PEO surfaces. Less plaque seemed to collect on these surfaces compared to adjacent normal tooth surfaces. Almost no plaque accumulated on the hydrophobic crown surface and it was virtually devoid of stainable pellicle. Even after 7 days in the mouth without oral hygiene this surface was very hydrophobic and the disclosing solution could not spread.     

Plaque pH measurements by different methods on the buccal and approximal surfaces of human teeth after a sucrose rinse

Changes in plaque pH after a sucrose rinse were simultaneously monitored by plaque sampling, touching of the plaque with antimony and glass electrodes, and telemetry. The minimum pH at approximal sites was approximately 0.7 pH units lower than that on buccal surfaces. The pH at the approximal site remained below resting levels after 120 min, and the area under the pH response curves from this site was five times greater than that from the buccal surfaces.     

Presence of Streptococcus mutans and lactobacilli in saliva and on enamel, glass ionomer cement, and composite resin surfaces

The quantity of S. mutans, total streptococci, and lactobacilli on sound enamel surfaces and 1-yr-old glass ionomer cement and composite resin fillings with the cervical margins placed subgingivally was compared intra-individually. The amount of bacteria was compared to their number in saliva. The evaluation was done in a cross sectional study, where the patients continued to use their customary oral hygiene procedures and during a 14-day period of experimental plaque formation. The number of lactobacilli and S. mutans recovered from the test surfaces indicated that the critical salivary concentrations necessary for the isolation of S. mutans and lactobacilli from glass ionomer cement and composite resin surfaces are the same as for the enamel surfaces. The fluoride levels in plaque adjacent to glass ionomer cement will not become high enough to inhibit the accumulation of the investigated bacteria.     

Presence of Streptococcus mutans and lactobacilli in saliva and on enamel, glass ionomer cement, and composite resin surfaces

Abstract— The quantity of S. mutans , total streptococci, and lactobacilli on sound enamel surfaces and 1-yr-old glass ionomer cement and composite resin fillings with the cervical margins placed subgingivally was compared intraindividually. The amount of bacteria was compared to their number in saliva. The evaluation was done in a cross sectional study, where the patients continued to use their customary oral hygiene procedures and during a 14-day period of experimental plaque formation. The number of lactobacilli and S. mutans recovered from the test surfaces indicated that the critical salivary concentrations necessary for the isolation of S. mutans and lactobacilli from glass ionomer cement and composite resin surfaces are the same as for the enamel surfaces. The fluoride levels in plaque adjacent to glass ionomer cement will not become high enough to inhibit the accumulation of the investigated bacteria.     

Development of plaque on enamel

Intact human enamel was used to study the development of dental plaque. This procedure involved placing cylinders of surface enamel in an appliance that was worn by one subject up to seven days. By using a technique of slight demineralization of the enamel, pellicle and plaque could be sectioned along with the enamel for light and transmission microscopy. Scanning electron microscopic and microbiologic studies were also performed on the specimens. Microbiologic investigation of two day old samples from the one individual revealed the presence of streptococci and a smaller number of Neisseria. Streptococcus sanguis was the predominant streptococcal species. Early samples of dental plaque appeared to have more desquamated epithelial cells than seven day old plaques. Electron micrographs suggest that bacteria may digest these epithelial cells. An apparent distinction between bacteria adjacent to the enamel and bacteria adsorbed to the surface of the enamel was. noted. Adsorbed bacteria appeared to alter their morphology and excrete an extracellular material which later formed a pellicular layer. Seven day old samples characteristically showed an alignment of rods superior to the adsorbed cocci. The formation of dental plaque on enamel cylinders may better simulate "in vivo" conditions than previous techniques.     

Epithelial cells associated with the development of dental plaque.

Transmission and scanning electron microscopy were used to study desquamated epithelial cells in the developing and mature dental plaque. Epithelial cells were found attached to enamel as early as one hour after clean enamel cylinders were placed in the mouth, and they were seen in great numbers after two days. These cells generally had bacteria attached to their surfaces. Epithelial cells adsorbed to the surface of the enamel appeared to alter their morphology, and mineralization of these structures was also observed. Desquamated epithelial cells could play a role in the initial bacterial colonization of tooth enamel since it is known that bacteria selectively adsorb onto their surfaces. Hydroxyapatite may affect the morphologic alteration of epithelial cells adsorbed to tooth enamel. From the observation of developing and mature plaque samples, it seems that certain suprasurface pelliclelike structures might be epithelial cells.     

Effect of a hydroxyl ion-releasing composite resin on plaque acidogenicity

The aim of this in vivo study was to evaluate the neutralizing capacity, registered as change of plaque acidogenicity, on aged proximal restorations of an ion-releasing composite resin (IRCR), which releases hydroxyl, calcium, and fluoride ions at low pH. Twenty patients, with a mean age of 63 years (range 43-85), participated. All had one aged proximal IRCR restoration (mean age 15 months) and one nonrestored enamel surface to make an intraindividual comparison possible. The neutralizing effect of the IRCR was evaluated by measuring plaque pH, using the microtouch method, after a mouthrinse with 10% sucrose. The plaque pH measurements were repeated 1.5 years later on the IRCR (mean age 34 months), the enamel surfaces and a universal hybrid composite resin (CR). At both 15 and 34 months, the plaque on the IRCR surfaces showed the least acidogenic potential for the whole 60-min time interval. The largest differences between the IRCR, CR and enamel were found during the first 15 min. At 15 months, the total areas under the plaque pH curve (AUC(5.7) and AUC(6.2)) differed significantly between the IRCR and enamel surfaces for the time periods 0-5 min and 5-15 min. At 34 months, significant differences were found between IRCR and CR at the 0- to 5-min time period. It can be concluded that IRCR restorations countered the plaque pH fall and maintained it at levels where less enamel and dentin demineralization can occur.     

The potential for dental plaque to protect against erosion using an in vivo-in vitro model--a pilot study

BACKGROUND: Tooth erosion is a problem for professional wine tasters (exogenous erosion from frequent exposure to wine acids) and for people with gastro oesophageal reflux disease (GORD) and bulimia who experience frequent reflux of gastric contents into the mouth (endogenous erosion from mainly HCl). The objective in this study was to determine whether plaque/pellicle could provide teeth with any protection from two common erosive acids, using an in vivo-in vitro technique. METHODS: Tiles of human tooth enamel and root surfaces were prepared from six extracted, unerupted third molar teeth and sterilized. Mandibular stents were prepared for six volunteer subjects and the tiles bonded to the buccal flanges of these stents. They were worn initially for three days to permit a layer of pellicle and plaque to form over the tile surfaces, and for a further 10 days of experimentation. Following cleaning of the plaque/ pellicle layer from the tiles on the right side flange, all the tiles were submerged in either 0.06M HCl or white wine for an accumulated time of 600 and 1500 minutes, respectively. Depths of erosion were determined using light microscopy of sections of the enamel and root tiles. SEM of the lesion surfaces was carried out to investigate the nature of erosive damage and of plaque/pellicle remnants. RESULTS: Retained plaque was found to significantly inhibit dental erosion on enamel, from contact with both HCl and wine, compared with that resulting following its removal. However, it was found to provide no significant protection on root surfaces. SEM analysis of the tile surfaces revealed marked etching of enamel on the cleaned surfaces, and considerable alteration to the appearance of remaining plaque and pellicle on most surfaces. CONCLUSION: Within the limitations of numbers of specimens, dental plaque/pellicle provided a significant level of protection to tooth enamel against dental erosion from simulated gastric acids and from white wine, using an in vivo-in vitro model. It was unable to provide any significant protection to root surfaces from these erosive agents. Possible reasons for this difference are explored.     

Scanning electron microscope study of early bacterial penetration of human enamel in initial caries

Bacterial penetration of enamel during initial manifestations of the carious process was studied in intact buccal enamel of 8 impacted third permanent molars. After cleaning them from organic plaque, each buccal plane was cut into five segments, one of them serving as a control specimen and the other four being fixed into slots on partial prostheses of our volunteers. The specimens were left in oral cavity for 7, 14, 21 and 28 days, where they were exposed to the action of cariogenic factors. After removal of the organic plaque, the specimens were broken in two and the bacterial penetration into enamel was observed on the fractured cross-section using scanning electron microscope. In 4 specimens from the group orally exposed during a 7-day period, individual coccoid or bacilliform bacteria were found to have penetrated 5-10 microns deep. The number of bacteria and the depth of their penetration into enamel increased with the duration of oral exposition. Among the specimens orally exposed during 28 days, bacteria were observed to be present both individually and in colonies, penetrating to the depth of 60-90 microns. These bacteria could quite easily communicate with saliva and plaque via the pore system, which allowed them to produce metabolites, including lactic acids. This, in turn, allowed them to penetrate through the enamel, thus altering both the course and rate of the carious process progression.     

Confocal laser scanning microscopic analysis of early plaque formed on resin composite and human enamel

The purpose of this study was to analyse quantitatively the early bacterial plaque formed on resin composite and human enamel in vivo, using a confocal laser scanning microscope. Test pieces of resin composite and human enamel were retained at the buccal surfaces of the upper first molars of three volunteers for 4, 8 and 24 h to allow plaque formation. Then, the specimens were immersed in propidium iodide in phosphate-buffered saline to stain adherent bacteria and observed with a confocal laser scanning microscope. The ratios of the area occupied by microorganisms to the whole area of the optical field were calculated using a photo-image analysis system. The thickness of the plaque was also measured. Quantitative analysis revealed that the resin composite showed significantly higher bacterial adherence than human enamel throughout the test period. A difference was noticed in the morphology of the bacteria between the two groups. Our findings suggest that resin composite shows higher bacteria adherence during early plaque formation compared with human enamel. In addition, the present findings may suggest a presence of the difference in bacterial composition of plaque in both specimens.     

Comparative study of plaque pH on enamel and cemental surfaces

This study was undertaken to compare the pH response of enamel and cemental plaques to a sucrose rinse challenge. The plaque pH readings on the enamel surfaces were consistently lower than the subjacent cemental plaque pH readings on the same teeth at the same time intervals following the sucrose exposure.     

Effects of calcium glycerophosphate and sodium fluoride on the induction in vivo of caries-like changes in human dental enamel.

Optical changes interpreted as demineralization on the enamel surfaces of 8 volunteers who did not clean their teeth for 18 days, while rinsing 9 times each day with 50 per cent sucrose solution, were not reduced by the addition of calcium glycerophosphate (1 per cent w/v) in the rinse. No changes were observed in plaque accumulation or in the plaque pH fall after sucrose attributable to calcium glycerophosphate. Topical applications of sodium fluoride (2 per cent) tended to inhibit the development of similarly-induced changes but the effect was not statistically significant with the small number of subjects.