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The antibacterial activity of the peptide PsVP-10 obtained from Pseudomonas sp. R10 against Streptococcus mutans and Streptococcus sobrinus was investigated. One hundred and twenty strains of S. mutans and 120 strains of S. sobrinus with and without glycocalyx were isolated from saliva samples in trypticase-yeast-cysteine-sucrose-bacitracin (TYCSB) agar. Bacterial identification was made by polymerase chain reaction. Glycocalyx production was observed in modified TYCSB agar and confirmed with a modified version of the microplate adherence assay. The minimum inhibitory concentration (MIC) of PsVP-10 bacteriocin was determined by means of the agar dilution method, and the time of bacterial death was calculated by means of colony-forming unit counts. The MIC of the bacteriocin PsVP-10 for both bacterial species with and without glycocalyx was < 2 mg/L and the time of bacterial death was less than 240 s for all the studied bacterial strains. Thus, bacteriocin PsVP-10 could be an interesting possibility to combat these cariogenic bacterial species.

Unlabelled: The objectives of this study were to evaluate and compare the amount and pattern of fluoride release from teeth after topical application of 2% NaF, 8% SnF2 and 1.23% APF at different time intervals. The growth inhibitory effects of this released fluoride ion was assessed on mutans streptococci (MS) and correlated with the fluoride release. Forty premolars divided into four groups were subjected to different topical fluoride treatments. All the teeth were immersed individually in deionized water and were transferred to containers at 1 hour, 1 day and 1 week time intervals. 240 samples in total were used for fluoride estimation by ion selective electrode method and the samples from the other subgroup were used for evaluation of antimicrobial activity on mutans streptococci (MS) by bacterial inhibition assay method. The results showed that the highest fluoride release (7.83 +/- 0.55 ppm) was seen in SnF2 treated specimens, as compared to that of NaF (3.71 +/- 0.60ppm) and APF (3.30 +/- 0.51ppm), the difference being statistically significant (P<0.01). This was observed immediately after 1 hour, followed by a drastic reduction thereafter. No zones of inhibition were observed at the released fluoride concentrations at different time intervals in the different groups., In Conclusion: 8% SnF2 is expected to have greater anticaries property from the high fluoride releasing property for prolonged period of time.

In natural ecosystems, micro-organisms grow preferentially attached to surfaces, forming matrix-enclosed biofilms. The aim of this study was to determine photodestruction levels in biofilms after subjecting them to photodynamic therapy. Biofilms of Streptococcus mutans, S. sobrinus, and S. sanguinis were grown on enamel slabs for 3, 5 or 7 d. Both the number of viable micro-organisms and the concentration of water-insoluble polysaccharides were analysed, and mineral loss (DeltaZ) analyses were performed on the enamel slabs. The antimicrobial potential of toluidine blue O (0.1 mg ml(-1)), associated with 85.7 J cm(-2) of a light-emission diode, was evaluated on the viability of 5-d biofilms. Both the number of micro-organisms and the concentration of water-insoluble polysaccharide increased with the age of the biofilms. A significant reduction ( approximately 95%) in viability was observed for S. mutans and S. sobrinus biofilms following photosensitization, with a > 99.9% reduction in the viability of S. sanguinis biofilms. In conclusion, a biofilm model was shown to be suitable for studying changes in bacterial numbers and enamel mineralization and for demonstrating the potential value of photosensitization in the control of in vitro biofilms.

This study aimed to determine whether the long-term use of a fluoride mouthrinse affects the salivary levels of mutans streptococci. Two hundred and fifteen schoolchildren (aged 9-10 years) participated. One hundred and forty-nine of these children had used a fluoride mouthrinse since 5 years of age at nursery school, and the remaining 66 children had not. DFT (decayed and filled teeth) was recorded, and the salivary levels of Streptococcus mutans and Streptococcus sobrinus were measured using mitis salivarius bacitracin agar. The group that had used a fluoride mouthrinse had a significantly lower prevalence of both S. mutans and S. sobrinus (p = 0.038) and a significantly lower DFT score (p < 0.001) than the other group. Using logistic regression analysis including caries experience at baseline as a dependent variable, the odds ratio of carrying S. mutans alone was 8.0 (p = 0.066) and that of carrying both S. mutans and S. sobrinus was 16.5 (p = 0.022) in the group that had not used the fluoride mouthrinse. Children carrying both S. mutans and S. sobrinus had a higher caries incidence in 1 year than the others, with odds ratios of 5.73 (p = 0.067) in the group with a fluoride mouthrinse and 3.47 (p = 0.035) in the group without it. These results show that the long-term use of a fluoride mouthrinse is associated with reduced salivary levels of mutans streptococci and this bacterial reduction may partly contribute to the suppression of dental caries in children using a long-term fluoride mouthrinse.

Objective: To determine the physicochemical properties of the mutanase of Trichoderma harzianum isolated from China and to study the influence of mutanase on the adherence of oral Streptococci and the structure of oral biofilms., Methods: Six fungal strains belonging to Trichoderma were tested for mutanase production in the same cultural condition, the strain producing the highest mutanase activity was studied further and the pH and temperature optimum of the enzyme was determined. The RT-PCR method was used to obtain the gene coding for mutanase and the product was cloned to pMD18-T simple vector for sequencing. Inhibition effects of mutanase on the adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 were studied by adherence test. The optical sectioning of biofilms with or without mutanase supplementation were analyzed by confocal laser scanning microscopy (CLSM)., Results: The highest enzymatic activity was achieved by Trichoderma harzianum Th1, the maximum activity was at pH 5.5 and at 40 degrees C. The nucleotide sequence was 92% homology with that of a known gene coding a mutanase (GenBank accession No. AJ243799). The adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 was significantly inhibited by mutanase. Compared with control, the biofilms with mutanase supplementation had lower height and sparser structure., Conclusions: The mutanase from Trichoderma harzianum Th1 can inhibit the adherence of oral Streptococci and had an influence on the structure of oral biofilms.

Although interactive forces, influenced by environmental conditions, between oral bacteria and tooth surfaces are important for the development of plaque, they have never been estimated. It is hypothesized that interactive forces, as measured by atomic force microscopy, between enamel with or without a pellicle and two strains of mutans streptococci become less attractive by the application of a Streptococcus mitis BMS biosurfactant coating. Upon approach of each of the strains toward bare and pellicle-coated enamel, adsorbed biosurfactant increased the range of the repulsive forces. Upon retraction of the enamel surface, small adhesion forces (0.8-0.9 nN) were measured for bare enamel that almost disappeared after biosurfactant coating. The prevalence and magnitude of the adhesion forces also decreased upon pellicle-coating of the enamel, with a minor effect of adsorbed biosurfactant. These findings indicate that adsorbed S. mitis BMS biosurfactant changes the interactive forces between the mutans streptococci studied and enamel, explaining the effects of biosurfactant on adhesion.

The purpose of this study was to evaluate the combined inhibitory effect of fluoride and xylitol on acid production by mutans streptococci, Streptococcus mutans NCTC10449 and Streptococcus sobrinus 6715, from glucose under strictly anaerobic conditions at fixed pH 5.5 and 7.0. The bacteria were grown in a tryptone-yeast extract broth under strictly anaerobic conditions (N2: 80%; H2: 10%; CO2: 10%). Reaction mixtures for acid production from glucose contained bacterial cells with fluoride (0-6.4 mM) and/or xylitol (60 mM). Acidic end products of glucose fermentation and intracellular glycolytic intermediates were assayed. The combination of fluoride and xylitol inhibited acid production more effectively than fluoride or xylitol alone. In the presence of fluoride and xylitol, the proportion of lactic acid in the total amount of acidic end products decreased, while the proportion of formic and acetic acids increased. Analyses of intracellular glycolytic intermediates revealed that xylitol inhibited the upper part of the glycolytic pathway, while fluoride inhibited the lower part. This study indicates that fluoride and xylitol together have synergistic inhibitory effects on the acid production of mutans streptococci and suggests that xylitol has the potential to enhance inhibitory effects of low concentrations of fluoride., (Copyright 2005 S. Karger AG, Basel.)

This study evaluated the antibacterial activity of the glass-ionomer cements Vitrebond (3M ESPE), Ketac Molar (3M ESPE) and Fuji IX (GC America) against S mutans, S sobrinus, L acidophilus and A viscosus, using the agar diffusion test. Inocula were obtained by the seed of indicators cultures in BHI broth incubated at 37 degrees C for 24 hours. Base layers containing 15 mL of BHI agar and 300 microL of each bacteria suspension were prepared in Petri dishes. Six wells measuring 4 mm in diameter were made in each plate and completely filled with one of the testing materials. A 0.2% chlorhexidine solution applied in round filter papers was used as control. Tests were performed 12 times for each material and bacteria strain. After incubation of the plates at 37 degrees C for 24 hours, the zones of bacterial growth inhibition around the wells were measured. Overall, the results showed the following sequence of antibacterial activity: Vitrebond (despite the activation mode) > 0.2% chlorhexidine > Ketac Molar > Fuji IX, according to Kruskal-Wallis and Mann-Whitney statistical tests. This study confirmed significant antibacterial activity for two conventional glass-ionomers and one resin-modified glass-ionomer material. The resin-modified glass-ionomer cement Vitrebond, regardless of the activation mode, presented the best antibacterial activity against S mutans and S sobrinus. The antibacterial activity against A viscosus for Vitrebond was similar to 0.2% chlorhexidine, while light activation reduced its antibacterial activity against L acidophilus.

The purpose of this study was to investigate the antibacterial activity of MMA/TBB resin containing newly developed amphiphilic lipids. The amphiphilic lipids, C10-L-Ala/pts and C12-L-Ala/pts, synthesized from the reaction of n-alkyl alcohol and L-alanine were dissolved in MMA at concentrations of 0.5, 1.0, 1.5, and 2.0 mol%. Resin mixtures of PMMA powder and each MMA liquid containing lipid and TBB were prepared for all tests. Both lipids gave antibacterial effect to MMA/ TBB resin. The addition of C12-L-Ala/pts to MMA resulted in a significantly higher antibacterial activity than the addition of C10-L-Ala/pts. In terms of bond strength, the bond strength of MMA/TBB resin to bovine dentin was significantly decreased by the addition of amphiphilic lipids. But for enamel, the bond strength of MMA/TBB resin with amphiphilic lipids was clinically acceptable for orthodontic brackets. In conclusion, amphiphilic lipids will be useful as a component of adhesive resin to give the latter an antibacterial effect.

Objectives: Resilient denture-lining materials are more susceptible to microbial adhesion than are denture base acrylic resin. As denture hygiene is essential to maintain the serviceability of the denture, disinfectant solutions have been suggested as a method for effectively cleaning resilient liners. The aim of this study was to determine the effectiveness of four different disinfectant solutions on four different resilient lining materials contaminated with Staphylococcus aureus, Streptococcus sobrinus, and Candida albicans., Method and Materials: Three microorganisms (Sta aureus, Str sobrinus, and C albicans), four resilient lining materials (Tempo, Immediate, Flexacryl soft, Ufi Gel P), and four disinfectant solutions (5.25% sodium hypochlorite, 2% sodium hypochlorite, 5% Deconex, and 3.5% Savlex solutions) were investigated. The four resilient lining materials were contaminated with each of the microorganisms, then the disinfection tests were applied to detect the efficacy of disinfectant solutions on resilient denture base materials in an in vitro test. The specimens (n = 5, total = 240) were soaked for 5 minutes in each solution to disinfect the microorganisms. The control specimens (one of each material; n = 5, total = 60) were soaked in distilled water for 5 minutes. As a result, a reduction in microorganism counts was determined. The Kruskal-Wallis and one-way analysis of variance tests were used to analyze for significant differences among test groups at the level of significance., Results: For all microorganisms, soaking in 5.25% sodium hypochlorite reduced the number of viable adherent microorganisms significantly compared to soaking in 2% sodium hypochlorite, which led to greater reduction than soaking either 5% Deconex or 3.5% Savlex. The use of 5.25% sodium hypochlorite in all groups is statistically significant., Conclusion: Disinfection of four different resilient denture-lining materials with 5.25% sodium hypochlorite solution was the most effective immersion technique. To achieve an efficient disinfection, the immersion time of the other disinfectant solutions could be increased.

Purpose: To evaluate the effect of 1.23% acidulated phosphate fluoride (APF) and the subsequent polishing on the in vitro adherence of mutans streptococci to resin composites in a model that simulated the oral cavity conditions., Methods: Samples of light-curing resin-based composites (Z100 and Z250) were treated with 1.23% APF gel (Sultan) for 1 minute and polished subsequently with Sof-Lex disks. The bacterial adherence of Streptococcus mutans and Streptococcus sobrinus to the APF gel treated resin samples and the subsequent polished samples were analyzed and compared with the adherence to the non-treated controls., Results: The 1.23% APF gel caused a significant increase in the adherence of Streptococcus mutans and Streptococcus sobrinus. The increasing rate of the bacterial adherence to resin composites after APF treatment ranged from 73.5% to 167% and differed according to the bacterial strains. A decrease in the adherence of the bacteria was found after subsequent polishing with Sof-Lex disks.

Aim: To investigate the use of polyol-containing chewing gums in a day-care centre (kindergarten) setting as a means to affect the growth of mutans streptococci and dental plaque., Design: Over a period of six months, 123 five-year-old children chewed xylitol (X group), sorbitol (G group), or did not chew gum (C group). Consumption of xylitol, and sorbitol was 4.5 to 5.0 g per day and subjects consumed in five supervised daily chewing episodes four at the day-care centres and one at home., Methods: Interproximal dental plaque was sampled at baseline and after six months for a laboratory study of mutans streptococci counts. The Quigley & Hein plaque index procedure was used. Interviews and questionnaires elucidated the acceptability of the programme., Results: Parents and kindergarten personnel regarded the programme as an important, additional procedure to promote better oral health. The children regarded the use of chewing gum as a pleasurable experience. Compared with groups G and C, there was a statistically significant reduction of mutans streptococci in the interproximal plaque in the X group. The Quigley & Hein plaque index scores tended to decrease in the X group, while no such trend was observed in the G group., Conclusions: Habitual use of relatively small daily quantities of polyol-containing chewing gum by young children may be regarded as an important additional caries-preventive procedure in a combined day-care centre and home setting. Especially xylitol-containing chewing gum may significantly reduce the growth of mutans streptococci and dental plaque which may be associated with dental caries.

We have previously demonstrated that a linalool-rich essential oil from Croton cajucara Benth presents leishmanicidal activity. In the present study, we demonstrate that this essential oil inhibits the growth of reference samples of Candida albicans, Lactobacillus casei, Staphylococcus aureus, Streptococcus sobrinus, Porphyromonas gingivalis and Streptococcus mutans cell suspensions, all of them associated with oral cavity disease. The purified linalool fraction was only inhibitory for C. albicans. Microbes of saliva specimens from human individuals with fixed orthodontic appliances, as well as the reference strains, were used to construct an artificial biofilm which was exposed to linalool or to the essential oil. As in microbial suspensions, the essential oil was toxic for all the microorganisms, while the purified linalool fraction mainly inhibited the growth of C. albicans. The compounds of the essential oil were separated by thin layer chromatography and exposed to the above-cited microorganisms. In this analysis, the proliferation of the bacterial cells was inhibited by still uncharacterized molecules, and linalool was confirmed as the antifungal component of the essential oil. The effects of linalool on the cell biology of C. albicans were evaluated by electron microscopy, which showed that linalool induced a reduction in cell size and abnormal germination. Neither the crude essential oil nor the purified linalool fraction is toxic to mammalian cells, which suggests that the essential oil or its purified components may be useful to control the microbial population in patients with fixed orthodontic appliances.

Dental biofilm harbouring oral bacteria is highly correlated with the progression of dental diseases. Disruption of biofilm formation via anti-adhesion agents is an alternative means to the antibacterial approach. Previous studies have shown that high molecular weight non-dialysable material (NDM) derived from cranberry juice inhibits the adhesion of Escherichia coli and the coaggregation of a variety of oral bacteria. In addition, it inhibits the formation of glucans and fructans synthesised by GTF and FTF. In the present study, we examined the anti-adhesion effect of NDM on S. sobrinus. NDM promoted desorption of S. sobrinus from biofilm in the presence and absence of extracellular glucans and fructans, although the effect was more pronounced in the absence of these polysaccharides. Precoating of the bacteria with NDM reduced their ability to form biofilm. Our results indicate that NDM could be exploited as an anti-biofilm agent.

The influence of dentin on the effectiveness of three antibacterial agents (triclosan, glutaraldehyde, NaOCl) on Streptococcus mutans, S. sobrinus, and Lactobacillus acidophilus was tested using the agar diffusion method with and without bovine dentin discs (200 mum and 500 mum thickness) placed between bacteria and test substances. The effect of 0.3% triclosan on all tester strains (100%) was reduced after passage through 500 mum dentin discs to 0% (L. acidophilus) and to 22% and 28% (S. mutans and S. sobrinus). Seal&Protect (Dentsply, Konstanz, Germany), a triclosan containing dental bonding agent, produced inhibition zones only against S. mutans, but no zone when applied on 200 mum dentin discs. The inhibition zones for 1% NaOCl and 5% glutaraldehyde against all tester strains were significantly increased up to 230% (glutaraldehyde) and 236% (NaOCl) when applied on dentin discs, compared to direct application (100%). Dentin may either decrease or increase the inhibitory effect of antibacterial agents.

Background: Previous studies have shown that high molecular-weight non-dialysable material derived from cranberry juice (NDM) inhibits co-aggregation of a variety of oral bacteria., Objectives: In the present study, we examined the effect of NDM on several constituents of the dental biofilm, glucosyltransferase (GTF) and fructosyltransferase (FTF), as well as on the adhesion of Streptococcus sobrinus., Results: The activity of immobilized and soluble GTF and FTF was inhibited by NDM (P > 0.05). NDM also inhibited adhesion of S. sobrinus to hydroxyapatite (P < 0.05)., Conclusions: Our results indicate that NDM may affect biofilm formation. One of the proposed mechanisms is via inhibition of extracellular polysaccharide synthesis, which promote the sucrose-dependent adhesion of oral bacteria as S. sobrinus.

Dentin bonding agents with antibacterial effects may inhibit secondary caries formation and pulp inflammation by eliminating residual bacteria in and on dentin. Therefore, the antibacterial effects of Prime & Bond NT (PB), Prime & Bond NT without fluoride (PBNF), Gluma Comfort Bond (GL), ABF, Xeno CF II (XE), 2-hydroxyethyl methacrylate (HEMA), triethylene glycol dimethacrylate (TEG-DMA), and 0.2% chlorhexidine were tested against Streptococcus mutans, S. sobrinus, and Lactobacillus acidophilus using the agar-diffusion method with and without bovine-dentin disks (200 microm and 500 microm thickness) placed between the bacteria and the test substances. Without dentin, ABF Primer showed growth inhibition for all bacterial strains. XE inhibited S. mutans and S. sobrinus, and PB S. sobrinus. ABF Bonding inhibited L. acidophilus. PBNF, HEMA, and TEGDMA did not have any antibacterial effects. Dentin disks of 500 microm thickness reduced the inhibitory effect of chlorhexidine to 23% to 54% compared with direct application. ABF Primer (nonpolymerized) produced inhibition zones against all tester strains regardless of dentin disks interposed or not. XE (against S. mutans and S. sobrinus) and PB (against S. sobrinus) did not produce any inhibition zones on 200 microm thick dentin. After polymerization, the ABF system did not inhibit bacterial growth on 200 microm thick dentin disks. A dentin barrier reduces significantly the antibacterial activity of chlorhexidine and dentin bonding agents.

Previous investigations showed that a high molecular mass, non-dialyzable material (NDM) from cranberries inhibits the adhesion of a number of bacterial species and prevents the co-aggregation of many oral bacterial pairs. In the present study we determined the effect of mouthwash supplemented with NDM on oral hygiene. Following 6 weeks of daily usage of cranberry-containing mouthwash by an experimental group (n = 29), we found that salivary mutans streptococci count as well as the total bacterial count were reduced significantly (ANOVA, P < 0.01) compared with those of the control (n = 30) using placebo mouthwash. No change in the plaque and gingival indices was observed. In vitro, the cranberry constituent inhibited the adhesion of Streptococcus sobrinus to saliva-coated hydroxyapatite. The data suggest that the ability to reduce mutans streptococci counts in vivo is due to the anti-adhesion activity of the cranberry constituent.

Cariogenic bacteria and periodontopathic bacteria are present in dental plaque as biofilms. In this study, we investigated the antibacterial effects of essential oils on the following oral bacteria: Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Streptococcus mutans, and Streptococcus sobrinus. We tested manuka oil, tea tree oil, eucalyptus oil, lavandula oil, and romarinus oil and determined their minimum inhibitory concentration and minimum bactericidal concentration. The essential oils inhibited the growth of the bacteria tested, manuka oil being the most effective. Minimum bactericidal concentration values showed that lavandula oil acts bacteriostatically, and the remaining oils, bactericidally. Periodontopathic bacterial strains tested were killed completely by exposure for 30 s to 0.2% manuka oil, tea tree oil or eucalyptus oil. Tea tree oil and manuka oil showed significant adhesion-inhibiting activity against P. gingivalis. All the essential oils tested inhibited the adhesion of S. mutans. This study showed that, among the essential oils tested, manuka oil and tea tree oil in particular had strong antibacterial activity against periodontopathic and cariogenic bacteria. From the viewpoint of safety, we also examined the effects of these essential oils on cultured human umbilical vein endothelial cells and found that, at a concentration of 0.2%, they had little effect on cultured cells.

We examined the antimicrobial effects of human beta-defensin-2 (hBD-2) on 17 species of oral streptococci to investigate the involvement of antimicrobial peptide activity in oral microflora development and the clinical use of the antimicrobial peptide for oral microflora control. Oral streptococci exhibit diverse levels of susceptibility to human beta-defensin-2 (hBD-2). Two major cariogenic bacterial species, Streptococcus mutans ( S. mutans) and S. sobrinus, were found to be susceptible to the peptide, indicating that it is a potential therapeutic agent for preventing dental caries. S. mitis exhibited the lowest susceptibility to the peptide. S. mitis is a major indigenous bacterium in the oral microflora, and our results suggest that it might possess a certain resistance mechanism against hBD-2.

The aim of this study was to investigate the in vitro effects of low concentrations of chlorhexidine (CHX) on the proliferation of Streptococcus sobrinus (ATCC 33478) and primary human gingival fibroblasts (HGF). Liquid cultures of bacteria or human gingival fibroblasts were exposed to CHX concentrations ranging from 0.07 to 40 microM in microtiter plates at 37 degrees C for 24 h. Bacteria or cells grown without CHX served as controls. The effects of CHX were determined either by measurements of the optical density (OD) of bacterial cultures or by evaluation of cell growth with the DNA-intercalating fluorescent stain H33342 in comparison to untreated controls. Results were evaluated calculating means and standard deviations. Data were statistically analyzed by an ANOVA using Post Hoc tests ( p<0.005). No growth inhibition of S. sobrinus was found at concentrations between 0.07 and 0.15 microM CHX, whereas 0.3 microM CHX led to an elongated (2 h more) lag phase and 0.6 microM CHX induced a lag phase of 4 h more and a minor inclination of the curve in the log phase. Concentrations of CHX>/=1.25 microM completely inhibited growth of S. sobrinus. On the contrary, CHX showed no significant effect on growth of HGF at concentrations

Although chlorhexidine is one of the most efficacious antimicrobial agents used for the prevention of dental caries, side effects limit its application. The effects of gaegurin 6 (GGN6), an animal-derived cationic peptide, and its derivatives PTP6 and PTP12 on the growth of oral streptococci were investigated to assess the potential of these agents for use in the prevention of dental caries. The minimal inhibitory concentrations of the peptides for inhibition of the growth of oral streptococci (Streptococcus mutans , S. sobrinus, S. sanguis and S. gordonii) ranged from 1.2 to 8.2 muM. The peptides also exhibited marked synergistic antibacterial effects with chlorhexidine or xylitol. The most effective combinations (fractional inhibitory concentration index of 0.5) were xylitol with GGN6 against S. gordonii 10558 and chlorhexidine with either GGN6 or PTP6 against S. sobrinus OMZ-175. These results indicate that cationic peptides alone or in combination with chlorhexidine or xylitol might prove effective for the inhibition of the growth of cariogenic oral streptococci in situ., (Copyright 2003 S. Karger AG, Basel)

The in vitro activities of human beta-defensin 3 (hBD-3) alone or combined with lysozyme, metronidazole, amoxicillin, and chlorhexidine were investigated with the oral bacteria Streptococcus mutans, Streptococcus sanguinis, Streptococcus sobrinus, Lactobacillus acidophilus, Actinobacillus actinomycetemcomitans, and Porphyromonas gingivalis. hBD-3 showed bactericidal activity against all of the bacterial species tested. The bactericidal effect was enhanced when the peptide was used in combination with the antimicrobial agents mentioned above.

Data obtained from studies on the antimicrobial properties of bonding agents are the subject of controversy, probably because of methodological differences. This study compared two commonly used in vitro methods, the disc agar diffusion test and the well agar diffusion test. Agar plates were seeded with Streptococcus sobrinus, Lactobacillus gasseri, or Actinomyces naeslundii. For the well diffusion test, wells cut out of the agar were filled with the test material, and for the disc method, discs impregnated with the test material were applied to the agar; the discs and wells were both 9 mm in diameter. After incubation, measurements of the zones of inhibition showed little agreement between the two methods when bonding agents were tested; the mean differences (+/- sdiff) in the zones of inhibition between the methods were 0.7 +/- 3.4 mm (P = 0.40, one sample t-test against zero), 4.9 +/- 4.4 mm (P = 0.97), and 0.8 +/- 4.3 mm (P = 0.47) for S. sobrinus, L. gasseri, and A. naeslundii, respectively. Mean differences were less contrasting when chlorhexidine and pure components were tested (P < 0.05 for S. sobrinus and L. gasseri). These results indicate the need for a gold standard method to evaluate the antimicrobial properties of bonding agents.

Treatment of dental caries as an infectious disease will require a paradigm shift in the way dentists and other health care professionals approach prevention and management of the disease. Prevention of dental caries has relied upon patient cooperation and often requires significant lifestyle changes that are at best difficult to implement and maintain or at worst ignored. This paradigm shift in the etiology, prevention, and treatment of dental caries demands that clinicians redirect their energies and emphasis from the "surgical" approach to dental caries to a "medical" strategy that focuses on early (prenatal if possible) risk assessment of the mother and implementation of appropriate therapeutic intervention, including use of antimicrobials, risk assessment of infants at 6 months of age, and a reduction in the levels of caries-producing bacteria. This revolution in how dentists practice and think will require that they develop strategies and curriculum to "retrain" practicing dentists and to train current and future dental students and residents as well as other medical colleagues on the essentials of the paradigm shift.

Propolis, a resinous hive product secreted by Apis mellifera bees, has been shown to reduce the incidence of dental caries in rats. Several compounds, mainly polyphenolics, have been identified in propolis. Apigenin and tt-farnesol demonstrated biological activity against mutans streptococci. We determined here their effects, alone or in combination, on glucosyltransferase activity, biofilm viability, and development of caries in rats. Sprague-Dawley rats were infected with Streptococcus sobrinus 6715 and treated topically twice daily as follows: (1) tt-farnesol, (2) apigenin, (3) vehicle control, (4) fluoride, (5) apigenin +tt-farnesol, and (6) chlorhexidine. Apigenin (1.33 mM) inhibited the activity of glucosyltransferases in solution (90-95%) and on the surface of saliva-coated hydroxyapatite beads (35-58%); it was devoid of antibacterial activity. tt-Farnesol (1.33 mM) showed modest antibacterial activity against biofilms and its effects on glucosyltransferases were minimal. The incidence of smooth-surface caries was significantly reduced by apigenin +tt-farnesol (60%), fluoride (70%), and chlorhexidine (72%) treatments compared to control (P < 0.05).

The effect of chewable saliva-stimulants on Streptococcus mutans levels in dental plaque and paraffin-stimulated whole saliva among participants who were mentally disabled was investigated. Over 64-days, 98 participants chewed one of four saliva-stimulating tablets five times/day. The tablets contained one of the following: xylitol (X) or sorbitol (S), or 1:1 mixtures of xylitol and erythritol (XE) or sorbitol and erythritol (SE). Consumption of xylitol and sorbitol in Groups X and S was 5.4 grams/day/ subject, and of each polyol in Groups XE and SE, consumption was 2.7 g/day/subject. Interproximal dental plaque and stimulated whole saliva were sampled at baseline, at Day 36, and Day 64. There was a statistically significant reduction of S. mutans in plaque and saliva counts in Groups X and XE. The percentage of S. mutans in total streptococci increased significantly in dental plaque in Group S but decreased in the other groups. The results suggest that xylitol-containing saliva stimulants may be more effective than sorbitol-containing products in controlling some caries-associated parameters in people who are mentally disabled. Also a relationship may exist between the pentitol-type xylitol and S. mutans, and erythritol may exert a specific biochemical effect on this organism, although further studies are needed.

Fluoride is used in dentistry as a prophylactic agent to reduce caries rates due to the demineralization/remineralization effect and its influence on the metabolism of cariogenic bacteria. The purpose of this study was to evaluate the cytotoxic effects of sodium fluoride (NaF) on three different cell lines and the antibacterial potency on Streptococcus sobrinus. Cell lines were treated with various concentrations of NaF ranging from 0.039 mM to 10 mM for 24 h. For microbial assays, concentrations of NaF between 0.03 mM and 10 mM were added to liquid cultures of bacteria. Our results showed that immortalized human keratinocytes (HaCaT) and human osteogenic sarcoma cells (SAOS-2) were similarly affected by concentrations up to 2.5 mM. However, cell growth of HaCaT was slightly more inhibited at 2.5 mM of NaF than SAOS-2. At concentrations between 0.62 mM and 10 mM, 3T3 mouse fibroblast cells reacted more sensitively than HaCaT and SAOS-2 to NaF. The 3T3 cells did not survive in the presence of 10 mM NaF. NaF caused no significant effect on all tested cells at concentrations of < or = 0.31 mM. NaF at 0.039 mM and 0.06 mM did not affect growth of S. sobrinus. At concentrations of 0.125 mM and 0.5 mM, growth was slightly reduced. The proliferation of S. sobrinus significantly decreased at 1 mM and 2 mM NaF. S. sobrinus survived at 4 mM, revealing a delayed log phase with a decreased proliferation. No viable S. sobrinus cells were detected at concentrations of > or = 8 mM NaF. Data analysis revealed that overall treatment effects were highly significant (P<0.05, analysis of variance, Tukey's difference test). This study indicates that cytotoxic effects due to NaF significantly vary in dependence upon the applied cell line. The toxicity of NaF approached 50% (TC50) at concentrations of 6 mM for HaCaT, 2.3 mM for 3T3 cells, and 7.5 mM for SAOS-2. Additionally, NaF revealed antimicrobial effects only at concentrations that are significantly higher than oral fluoride concentrations.

We evaluated the influence of molecular mass and degree of deacetylation of chitosan on the adsorption of Streptococcus sobrinus 6715 to saliva-treated hydroxyapatite (S-HA) by measuring the optical density of the bacterial cell suspensions released from saliva-treated hydroxyapatite. Twenty-five chitosan samples with different molecular masses (0.8-6 kDa) and degrees of deacetylation (10-95%) were prepared for the study. We found that the inhibition of adsorption of S. sobrinus 6715 to S-HA correlated positively with the molecular mass of chitosan (R = 0.876) and that the optimal degree of deacetylation was 50-60% for maximum inhibition of bacterial binding to S-HA. We also examined the effect of chitosan on zeta potentials of the oral bacteria and their surface hydrophobicities. It was observed that chitosan reduced the magnitude of the zeta potential and surface hydrophobicities of the oral bacteria. Thus, the results demonstrated that chitosan with a molecular mass of 5-6 kDa and a degree of deacetylation of 50-60% might have the potential to act as an effective anti-plaque agent because of its polycationic properties.

Representative strains of Streptococcus mutans and Streptococcus sobrinus showed differences in susceptibility to members of the monobactam group of beta-lactam antibiotics: S. sobrinus was less sensitive than S. mutans. The minimum inhibitory concentrations of aztreonam (AZT) and carumonam, both of which belong to this group, were 2,000 microg/ml for S. sobrinus and 125 microg/ml for S. mutans. Further addition of fosfomycin, bacitracin and sodium chloride to Mitis Salivarius agar (MS) supplemented with AZT resulted in growth inhibition of S. mutans and oral streptococci other than S. sobrinus, and was therefore used as a selective medium for S. sobrinus (MS-SOB medium). The average growth recovery of laboratory and clinically isolated strains of S. sobrinus on MS-SOB medium was 74.1% compared to that on MS medium. Seventy-eight percent of clinical samples in which S. sobrinus was detected yielded pure growth of S. sobrinus on MS-SOB medium., (Copyright 2002 S. Karger AG, Basel)

Long-term xylitol consumption leads to the emergence of xylitol-resistant (X-R) mutans streptococci. The aim of the present study was to compare cariogenic traits in X-R and xylitol-sensitive (X-S) strains. Six strains of mutans streptococci, three X-R and three X-S strains, were studied. Xylitol resistance and sensitivity were confirmed by growth in xylitol-supplemented media. Acid production from glucose or fructose or uptake of xylitol was initiated by adding (14)C-labelled glucose, fructose or xylitol to bacterial suspensions. The resultant metabolites were identified by HPLC. Lactate was the major metabolite from glucose, whether the bacteria were grown in the presence or the absence of xylitol. Lactate production per colony-forming unit was lower in X-S cells than in X-R cells. Fructose was metabolized by both X-R and X-S cells. Both X-R and X-S cells took up xylitol, but xylitol-5-P was detected in X-S cells only. Total polysaccharides were measured through production of C(14)-labelled ethanol-insoluble polymers from [U(14)-C]-sucrose. No difference in polysaccharide production was found between X-R and X-S cells. The present study thus does not support the contention that X-R are less cariogenic than X-S mutans streptococci.

Background: The authors examined the effect of xylitol, a naturally occurring sweetener, on levels of Streptococcus mutans and S. sobrinus. They also investigated xylitol's mechanism of action., Methods: The authors compared cariogenic bacteria levels before and after exposure to xylitol products in children and adults. In the first study, 187 children received xylitol-containing snacks in school for four weeks. In the second study, two adults received xylitol candy for four weeks. Unstimulated saliva samples were taken from all subjects. Gingival samples also were taken from the adults. The authors plated the samples on selective microbiological media. Individual isolates were plated on media with varying concentrations of xylitol, and were identified using specific DNA probes. Genetic relatedness was determined via pulse-field gel electrophoresis., Results: The children's salivary S. mutans levels remained stable before and after xylitol exposure. Further analysis of the S. mutans isolates was conducted for seven children. Bacteria from five of these children grew with 10 percent or less xylitol at baseline, while the bacteria from all seven children grew with 15 percent xylitol after exposure to the xylitol-containing snacks, suggesting that the S. mutans increased in tolerance to xylitol during exposure. Six children had isolates with the same genotype at both time points. S. mutans and S. sobrinus levels were reduced in one of the adults as a result of xylitol exposure, and the bacterial isolates became more xylitol tolerant. In the second adult, S. mutans and S. sobrinus levels increased, while the subject maintained the same proportion of susceptible and tolerant strains as that at baseline., Conclusions: Overall, consumption of xylitol-containing snacks and candy did not reduce S. mutans levels. However, bacteria from five children and one adult became more xylitol tolerant., Clinical Implications: These results provide a basis on which xylitol-containing products can be recommended and xylitol's mechanism of action can be explained to patients.

Tooth decay is an infectious disease caused by bacteria immobilized on the tooth surfaces. Eradication of these bacteria, for example Streptococcus sobrinus (S. sobrinus), from the oral cavity is essential in the prevention and treatment of tooth decay. We have tested the antimicrobial effect of several paraben derivatives such as methyl (MP), ethyl (EP), propyl (PP) and butyl (BP) against immobilized and planktonic S. sobrinus. The antibacterial effect was as follows: MP>EP>PP=BP on immobilized bacteria and MP>EP=PP>BP on planktonic bacteria. An antibacterial synergistic effect was found between several combinations of parabens on immobilized and planktonic S. sobrinus. Our results indicate that parabens are potential antibacterial agents against immobilized or planktonic bacteria found in the oral cavity.

The cariogenic potential of lactosylfructoside [O-beta-D-galactopyranosyl-(1-->4)-O-alpha-D-glucopyranosyl-(1<-->2)-beta-D-fructofuranoside] was estimated by experiments on oral streptococci in vitro and human dental plaque in situ. Lactosylfructoside was unable to support growth of the strains of Streptococcus mutans and S. sobrinus used in this study. However, it was able to support growth of strains of S. sanguis, S. mitis and S. oralis. Acid was produced rapidly by cell suspensions of S. oralis ATCC 10557 incubated with lactosylfructoside. Application of 5% w/v solution decreased the pH of human dental plaque. The minimum pH value was below 5.3. The results suggest that lactosylfructoside is as acidogenic as lactose and could be cariogenic if it is consumed frequently and retained for a long period in the mouth.

The decline in prevalence of dental caries in some segments of the population has been attributed mainly to extensive exposure to fluoride. Over the past decades, the use of fluoridated products has increased. During the same period, the consumption of food preservatives such as benzoates and sorbates has also increased substantially. Benzoates, in vitro, possess antibacterial properties similar to those of fluoride and in combination with fluoride could affect caries development. In the present study we explored the effects of sodium benzoate and fluoride in combination and alone on dental caries in our animal model. The results showed a combination of benzoate and fluoride reduced caries activity more effectively in rodents fed a cariogenic diet ad libitum than fluoride alone (p = 0.038).

Triclosan is a broad-spectrum hydrophobic antibacterial agent used in dermatological preparations and oral hygiene products. To gain further insight into the mode of action of Triclosan we examined its effects on membranes by performing leakage titrations of different oral bacteria and studying its interaction with model membranes through the use of different biophysical techniques. There was negligible efflux of intracellular material from Streptococcus sobrinus at the minimal inhibitory concentration of Triclosan; whatever leakage did occur commenced only at much higher concentrations. In contrast, no leakage was observed at even the minimal bactericidal concentration for Porphyromonas gingivalis. Triclosan decreased the onset temperature of the gel to liquid-crystalline phase transition of 1,2-dimyristoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-3-[phospho-rac-glycerol] membranes and was immiscible with these lipids in the fluid phase at concentrations greater than 5 mol%. Steady-state fluorescence anisotropy measurements of different phospholipid/Triclosan samples using 3-(p-6-phenyl-1,3,5-hexatrienyl)-phenylpropionic acid were consistent with the calorimetric data. Incorporation of increasing amounts of Triclosan into 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (DEPE) vesicles induced the nonlamellar H(II) hexagonal phase at low temperatures and new immiscible phases at temperatures below the main transition of DEPE. Taking these results together suggests that the antibacterial effects of Triclosan are mediated at least in part through its membranotropic effects, resulting in destabilized structures which compromise the functional integrity of cell membranes without inducing cell lysis., (Copyright 2001 Academic Press.)

Metal ions were evaluated as potential antimicrobial agents suitable for local delivery in the oral cavity for the treatment of periodontitis. Silver nitrate, copper chloride, and zinc chloride were tested for antimicrobial activity in in vitro killing assays conducted in phosphate buffered saline with a series of oral bacteria including gram-negative periodontal pathogens and gram-positive streptococci. Copper and zinc salts failed to exhibit strong and consistent activity against periodontal pathogens. In contrast, silver at a concentration of 0.5 microg/mL produced a 3 log10 reduction in colony forming units (CFU)/mL or greater against all periodontal pathogens tested including Porphyromonas gingivalis, Prevotella intermedia, Prevotella denticola, Bacteroides forsythus, Fusobacterium nucleatum vincentii, Campylobacter gracilis, Campylobacter rectus, Eikenella corrodens, and Actinobacillus actinomycetemcomitans. In comparison, substantially higher concentrations of silver nitrate failed to kill oral streptococci. A silver nitrate concentration of 25 microg/mL produced log10 reductions in CFU/mL of 3.5-5 in killing assays performed in human serum against P. gingivalis, demonstrating the ability of silver to retain activity in a biological medium similar to that encountered in vivo in the periodontal pocket. These results identify silver nitrate, an antimicrobial that may possess advantages over traditional antibiotics, as a potential agent for controlled release local delivery in the oral cavity for the treatment of periodontitis.

The antimicrobial and cytotoxic effects of sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were evaluated and compared in vitro. The minimal inhibitory concentration and minimal bactericidal concentration of NaOCl and NaDCC were tested for Streptococcus sobrinus, Streptococcus salivarius, Enterococcus faecalis, and Streptococcus mutans. The cytotoxic effect was assessed by using human fibroblast tissue culture. Survival rate was assessed by a protein determination method. Results showed that the minimal inhibitory concentration and minimal bactericidal concentration values of NaOCl and NaDCC for the tested bacteria were in a similar range. NaDCC in concentrations higher than 0.02%, and NaOCl in concentrations higher than 0.01% were lethal to fibroblasts. In conclusion it seems that both agents were very effective in killing bacteria, and their cytotoxicity to fibroblasts in tissue culture was similar.

The effect of 2-month usage of saliva-stimulating pastils containing either erythritol or xylitol was studied in a cohort of 30 subjects assigned to the respective polyol groups (n = 15). The daily consumption level of both polyols was 5.2 g, used in 5 daily chewing episodes. The mean weight of total plaque mass (collectable during a standard period of 3 min from all available tooth surfaces) was reduced significantly in the xylitol-group, while no such effect was observed in the erythritol-group. This reduction in plaque mass was accompanied by a significant reduction in the turbidity readings (A(660)) of aqueous plaque suspensions; no such effect was observed in the erythritol-group. However, plaque protein levels did not differ between baseline and endpoint in either polyol group. The plaque and salivary levels of Streptococcus mutans and plaque levels of total streptococci were reduced significantly in the xylitol-group, while no such effect was detected in the erythritol-group. However, either polyol regimen had no effect on plaque levels of S. sobrinus. The results suggest that systematic use of xylitol-containing saliva stimulants may be more effective in controlling some oral-hygiene-related and caries-associated parameters than similar use of erythritol-containing products. The results also speak for a special relationship between xylitol and S. mutans. However, owing to the great potential of erythritol as a caries-reducing agent -- based on the tetritol nature of erythritol -- the present laboratory results should be considered preliminary and subject to verifying clinical studies.

This study examined the antibacterial activities of the bonding systems Syntac, EBS and Scotchbond 1, the polyacid-modified composite resins Hytac and Compoglass, and the composite resins Tetric, Z100 and Scalp-it. They were evaluated using the cariogenic bacteria Streptococcus mutans, Lactobacillus salivarius, Streptococcus sorbinus and Actinomyces viscosus in vitro with a modified cylinder drop plate agar diffusion assay. All adhesives of the dentin bonding systems and the polyacid-modified composite resins exhibited various degrees of antibacterial activity against all of the test bacteria. On the contrary, composite resins did not affect bacterial growth. The data suggest that the use of these adhesives and polyacid-modified composite resins may reduce the consequences of microleakage owing to their antibacterial properties.

The study of biofilm structure and function mandates the use of model systems for which a host of environmental variables can be rigorously controlled. We describe a model of supragingival plaque containing Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis wherein cells are cultivated anaerobically in a saliva-based medium on hydroxyapatite discs coated with a salivary pellicle, with material and pieces of apparatus common to all microbiology laboratories. After 0.5 hr, 16.5 hrs, 40.5 hrs, and 64.5 hrs, the composition of adherent biofilms was analyzed by culture techniques, live/dead fluorescence staining, and confocal laser scanning microscopy. Repeated independent trials demonstrated the repeatability of biofilm formation after 40.5 hrs and 64.5 hrs. Brief exposures of biofilms to chlorhexidine or Triclosan produced losses in viability similar to those observed in vivo. This biofilm model should prove very useful for pre-clinical testing of prospective anti-plaque agents at clinically relevant concentrations.

Xylitol possesses a unique property distinct from the other caries-preventive sweeteners. This sugar alcohol cannot be metabolized to acids but is taken up by Streptococcus mutans and accumulated as a toxic sugar-phosphate in the cells, resulting in growth inhibition. Due to the accumulation, xylitol induces biological responses including the emergence of xylitol-insensitive populations. Therefore, we expected another response induced by xylitol and found a new phenomenon, that cells repeatedly cultured in the presence of xylitol evolved into those exhibiting an elevated dextran-dependent aggregation phenotype. This phenotype was found to result from expression of the gbpC gene, which was previously reported to be expressed only under certain stress conditions. Construction of a Strep. mutans isogenic mutant carrying the gbpC::lacZ fusion gene indicated that gbpC expression of cells repeatedly cultured in the presence of xylitol was elevated 20-fold. DNA transfer experiments indicated that this phenotypic change did not appear to be due to a mutation. These cells also exhibited decreased adhesion to glass surfaces when grown in the presence of sucrose. This may be one of the ways by which some populations of Strep. mutans are removed from dental plaques.

The aims of this present study were (1) to assess the antimicrobial effect of ozone from a novel ozone-generating device (Heolozone, USA) [0.052% (v/v) in air delivered at a rate of 13.33 ml.s(-1)] on primary root carious lesions (PRCLs) and (2) to evaluate the efficacy of ozone specifically on Streptococcus mutans and Streptococcus sobrinus. In study 1, 40 soft PRCLs from freshly extracted teeth were randomly divided into two groups to test the antimicrobial effect on PRCLs from exposure to ozonated water for either 10 or 20 s. Half of a lesion was removed using a sterile excavator. Subsequently, the remaining lesion was exposed to the ozonised water for a period of either 10 or 20 s (corresponding to 0. 069 or 0.138 ml of ozone, respectively). Using paired Student t tests, a significant (p<0.001) reduction (mean +/- SE) was observed in the ozone-treated groups with either a 10-second (log(10) 3.57+/-0.37) or 20-second (log(10) 3.77+/-0.42) ozone application compared with the control groups (log(10) 5.91+/-0.15 and log(10) 6.18+/-0.21, respectively). In study 2, 40 sterile saliva-coated glass beads were randomly divided into two groups for each micro-organism. One glass bead was put into each bijou bottle with 3 ml of Todd-Hewitt broth. S. mutans and S. sobrinus were inoculated anaerobically overnight. Each glass bead was then washed with 2 ml of phosphate-buffered saline. Immediately, 10 s of ozone gas was applied to each glass bead in the test groups. There was a significant (p<0.0001) reduction (mean +/- SE) in ozone-treated samples for S. mutans (log(10) 1.01+/-0.27) and S. sobrinus (log(10) 1.09+/-0.36) compared with the control samples (log(10) 3.93+/-0.07 and log(10) 4.61+/-0.13, respectively). This treatment regime is an effective, quick, conservative and simple method to kill micro-organisms in PRCLs. Ozone gas application for a period of 10 s was also capable of reducing the numbers of S. mutans and S. sobrinus on saliva-coated glass beads in vitro.

Exogenously added polyphenoloxidase (EC 1.14.18.1), an enzyme which oxidizes tyrosine residues and is commonly found in many dietary components, abolished the aggregation of Streptococcus sobrinus 6715 by high-molecular-weight dextran. The enzyme decreased glucan-binding lectin and/or glucosyltransferase I activities.

Polycarboxylates, such as carboxymethylcellulose and hyaluronan, were found to be reversible inhibitors of the glucan-binding lectin of Streptococcus sobrinus. When the carboxylate groups were coupled to ethylenediamine, or reduced with carbodiimide-borohydride, inhibitory powers were lost. Similarly, N-deacetylated hyaluronan had poor inhibitory powers, probably due to the introduction of positive charges into the polymer. Other polymers, such as chondroitin sulfates, dextran sulfate, fetuin, heparin were not inhibitors. It appears that inhibition is based on repeating carboxylates, free of influence from ammonium groups. Such polymers have the property of complexing with metals. Earlier studies had concluded that the streptococcal lectin depended on manganese for activity. It is likely the carboxymethylcellulose and hyaluronan perturb essential metal coordination centers in the lectin. Polycarboxylates may have value in oral health care by acting on glucan-dependent microbial adhesion and biofilm formation.

Chocolate is suspected to contain some caries-inhibitory substances. The cariostatic activity of cacao mass extract (CM), the main component of chocolate, was examined in vitro and in experimental animals. CM showed no detectable effects on the cellular growth and acid production of mutans streptococci. On the other hand, the cell-surface hydrophobicity of mutans streptococci was significantly reduced by the presence of CM. Furthermore, insoluble glucan synthesis by the glucosyltransferases from either Streptococcus mutans MT8148R or Strep. sobrinus 6715 was inhibited by CM, but not significantly. Hence, the sucrose-dependent cell adherence of mutans streptococci was also depressed by CM. Finally, CM in both a 40% sucrose diet and drinking water resulted in reductions of caries development and plaque accumulation in rats infected with Strep. sobrinus 6715, but not significantly. These results indicate that cacao mass extract possesses some anticariogenic potential, but its anticaries activity is not strong enough to suppress significantly the cariogenic activity of sucrose.

Cacao bean husk extract (CBH) was examined for inhibitory effects on the caries-inducing properties of mutans streptococci in vitro and on caries development in specific pathogen-free Sprague-Dawley rats infected with mutans streptococci. CBH reduced the growth rate of almost all oral streptococci examined, which resulted in the reduction of acid production. Furthermore, insoluble glucan synthesis by the glucosyltransferases from Streptococcus mutans MT8148R and Streptococcus sobrinus 6715 was significantly inhibited by CBH. Hence, the sucrose-dependent cell adherence of mutans streptococci was also depressed by CBH. The administration of CBH in drinking water resulted in significant reductions of caries development and dental plaque accumulation in rats infected with either Strep. sobrinus 6715 or Strep. mutans MT8148R, and the minimum cariostatic concentration was 1.0 mg/ml. These results indicate that CBH possesses powerful anticariogenic potential.

The aim of our study was to investigate the effect of resin composite components on glucan synthesis by glucosyltransferase (GTase) derived from a cariogenic bacterium, Streptococcus sobrinus B13. The eluates from cured composites stored in 0.05 mol/L potassium phosphate buffer (KPB) (pH 6.8) for 2 weeks at 37 degrees C stimulated the formation of water-insoluble glucan, whereas those from amalgam inhibited it. This finding suggests that the eluates from the resin composites enhance GTase activity and contribute to plaque formation. In the individual resin components, a diglycidyl methacrylate and triethylene glycol dimethacrylate significantly enhanced GTase activity. However, most polymerization inhibitors and accelerators were potent inhibitors of enzyme activity. Thus, GTase stimulation by eluates from resin composites is hypothesized to be heavily dependent on the effect of the eluated resin monomers, even though the other eluting components had inhibitory effects on GTase., (Copyright 2000 John Wiley & Sons, Inc.)

Dental caries is a chronic infectious disease caused by the accumulation of bacterial plaque (biofilm) on tooth surfaces. Antibacterial agents, in addition to other preventive measures, can control dental plaque accumulation. Amine fluorides (AmF) are known anticaries agents for over 30 years. The purpose of our study was to assess the adsorption and desorption of AmF to experimental dental biofilm and to evaluate the effect of AmF on Streptococcus sobrinus 6715 and glucosyltransferase (GTF) activity in experimental dental biofilms. The experimental plaque model used in this study consists of hydroxyapatite beads coated with human saliva (sHA), followed by adsorption of S. sobrinus and synthesis of in situ polysaccharides. Our results show that the viability of S. sobrinus in biofilm decreased as the concentration of AmF and chlorhexidine (CHX) increased. The concentration of AmF and CHX required to kill S. sobrinus adherent to sHA is about 100 times greater than the concentration required to kill the same amount of planktonic bacteria. Adsorption of AmF to surfaces was more than 90% and the desorption of AmF from our experimental model was limited. Pre-adsorption of AmF on the surface increased adhesion of S. sobrinus but also resulted in surface killing of the adsorbed bacteria. At low concentrations AmF increased GTF activity in solution by about 10%, but at concentrations above 0.1 mM it inhibited GTF activity. Inhibition of GTF on the surface required about 100 times more AmF than in solution. Our results show that AmF retains its anticariogenic effects in solution and in biofilm systems.

Glucan-binding lectin (GBL) activity of Streptococcus sobrinus was significantly reduced by fluoride in the growth medium. Approximately 1.5 mM fluoride was required for a 50% reduction in GBL activity. In addition to the GBL, several other glucan-binding proteins were reduced when the bacteria were grown in subinhibitory fluoride. Fluoride had no effect on glucosyltransferases (GTFs), enzymes capable of converting sucrose into alpha-1,6-glucans. All the proteins were detected by use of enhanced chemiluminescence (ECL of fluorescein-labeled dextran) and Western blotting of renatured SDS-PAGE gels. The effects of fluoride on the bacteria were abrogated when the manganous ion was included in the growth medium. It thus appears that one mechanism of action of fluoridated water is its effects on glucan-binding proteins. The fluoride may be reducing metabolism of the mangano aquo ion, essential for expression of the glucan-binding proteins.