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Adherence of Streptococcus mutans to an E-glass fiber-reinforced composite and conventional restorative materials used in prosthetic dentistry.

The adherence of Streptococcus mutans to E-glass used in fiber-reinforced composites, denture base polymer, and four other restoratives was investigated. The materials were studied with and without a parotid saliva and serum pellicle. Specimens of the studied materials (E-glass, denture base polymer, titanium, cobalt-chromium alloy, gold alloy, and grained feldspar ceramic) were incubated in a suspension of S. mutans, allowing initial adhesion to occur. The degree of bacterial adhesion was studied using scanning electron microscopy (SEM). The studied uncoated materials showed rather similar adhesion of S. mutans. Saliva coating resulted in a decrease of adherence to all materials except glass. With a saliva pellicle E-glass showed the strongest ability to bind S. mutans, and it differed significantly from the other studied materials. Serum coating markedly decreased adhesion to all materials, and only minor differences among the studied materials were observed. The results of this study suggest that the studied restoratives are rather similar with respect to S. mutans adhesion and that a saliva pellicle may promote adhesion of S. mutans to glass fibers.

Bacterial Adhesion↗

Adsorption of parotid saliva proteins and adhesion of Streptococcus mutans ATCC 21752 to dental fiber-reinforced composites.

The use of fiber-reinforced composites (FRC) in dentistry has increased during recent years. In marginal areas of crowns and removable partial dentures the fibers may become exposed and come into contact with oral tissues, saliva, and microbes. To date, few articles have been published on oral microbial adhesion to FRCs. The aim of this study was to compare different FRCs, their components, and conventional restorative materials with respect to S. mutans ATCC 21752 adhesion and adsorption of specific S. mutans binding proteins. Surface roughness of the materials was also determined. Four different FRCs, a restorative composite, and a high-leucite ceramic material were studied. Polyethylene FRC was found to be significantly rougher than all other materials. Aramid FRC also showed higher surface roughness in comparison with all materials but polyethylene FRC. Without a saliva pellicle, adhesion of S. mutans coincided with surface roughness and polyethylene and aramid FRC promoted S. mutans adhesion better than the other smoother materials. In the presence of salivary pellicle, ceramic and polyethylene FRC bound more bacteria than the other materials studied. Higher quantities of S. mutans binding proteins in the pellicles may in part account for the higher S. mutans adhesion to saliva-coated ceramic and polyethylene FRC.

Acrylic Resins↗

Antibodies against Streptococcus mutans and glucosyltransferases in caries-free and caries-active military recruits.

1) Plaque samples from caries-active recruits showed a higher incidence of S. mutans than plaque samples from caries-free recruits. The persistance of the S. mutans serotypes in the individual plaque samples attested to the consistency of S. mutans in human dental plaque. 2) S. mutans serotype d is predominantly present in approximal plaque of caries-active subjects. 3) Serum antibody titers against S. mutans are higher in caries-active individuals. 4) Sera from caries-free or caries-active individuals inhibit the insoluble glucan synthesis by the cellbound GTF to the same extent. 5) It is unlikely that antibodies against S. mutans in parotid saliva or serum play a major role in the protection against dental caries in Dutch military recruits.

Adult↗

Growth and adsorption of Streptococcus mutans 6715-13 to hydroxyapatite in the presence of lactoferrin.

The growth of Streptococcus mutans 6715-13 in a rich medium (Todd Hewitt broth) was drastically reduced by addition of apo-lactoferrin (apo-Lf); this effect was bacteriostatic and reversible by saturation of Lf with iron. The influence of Lf, salivary proteins (SP) and bovine serum albumin (BSA) on the attachment of Streptococcus mutans to hydroxyapatite (HA) was successively investigated. Sorption of Lf, SP, and BSA to HA was dependent on the protein concentration and reached the end-point at about 80 mg of proteins per gram of HA. Similarly, the number of streptococci adsorbed to HA was correlated to the amount of cells available up to at least 10(7) cells per mg of HA. The adsorption of Lf, SP and BSA on HA reduced the number of attaching S. mutans cells. In particular, SP reduced the adsorption of S. mutans by 30%, whereas pre-coating of HA with apo- or iron-saturated Lf resulted in a three orders of magnitude reduction of S. mutans adsorption to HA, as demonstrated by means of different experimental procedures. The powerful adherence-inhibiting effect of apo-Lf together with its noticeable antibacterial activity towards S. mutans points to a biological significance of these phenomena also in vivo.

Adsorption↗

Streptococcus mutans and dental caries in humans: a bacteriological and immunological study.

Plaque samples from caries-active subjects showed a higher incidence of S. mutans than plaque samples from caries-free subjects. This was especially evident in approximal incisor plaque. S. mutans serotype d was almost exclusively present in approximal plaque obtained from caries-active subjects. Tooth surfaces infected with S. mutans still harbored this micro-organism 10 months later, while uninfected tooth surfaces remained free of S. mutans. Caries development predominantly occurs on those tooth surfaces which harbor relatively high percentages of S. mutans (> 5%). It is unlikely that serum or saliva antibodies against S. mutans play a major role in the protection against dental caries in these caries-free subjects since subjects with the greatest number of decayed surfaces showed the highest antibody titre as measured by haemagglutination or by the enzyme-linked immuno sorbent assay (ELISA).

Adolescent↗

Effect of water-soluble reduced chitosan on Streptococcus mutans, plaque regrowth and biofilm vitality.

The purpose of the present study was to examine the effects of a newly developed water-soluble reduced chitosan on Streptococcus mutans, plaque regrowth, and biofilm vitality. A 1.0%, water-soluble reduced chitosan, with pH ranging from 6.0 to 6.5, molecular weights between 3,000 and 5,000 Da, and 70% degree of deacetylation, was used. To determine antibacterial and antiplaque potency of chitosan, minimal inhibitory concentrations (MICs) for S. mutans and S. sanguinis (formerly S. sanguis), short-term exposure to S. mutans, and clinical trial of plaque regrowth and biofilm vitality were conducted. Twelve dental students volunteered to participate in the 6-week, double blind, randomized clinical trial using the classical 4-day plaque regrowth design. The MIC of water-soluble reduced chitosan for S. mutans was 1.25 g/l. While the cells exposed to distilled water (DW) grew rapidly, with a maximum turbidity reached by 16 h postinoculation, S. mutans exposed to chitosan (5.0 g/l) exhibited a substantial delay in growth and reached a maximum turbidity by 32 h postinoculation. The chitosan solution reduced the plaque index and the vitality of the plaque flora significantly when compared to DW, but this was less than the reductions found with the positive control of 0.1% chlorhexidine solution. The water-soluble reduced chitosan exhibited potent antibacterial effect on S. mutans, and displayed a significant antibacterial and plaque-reducing action during the 4-day plaque regrowth.

Adult↗

Characterization of Streptococcus mutans diversity by determining restriction fragment-length polymorphisms of the gtfB gene of isolates from 5-year-old children and their mothers.

The diversity between Streptococcus mutans clinical isolates from 5-year-old children and their mothers in two South African ethnic groups was investigated. The gtfB gene encoding for glucosyltransferase (EC 2.4.1.5), an enzyme responsible for the synthesis of extracellular polysaccharides was characterized by PCR-RFLP with HaeIII restriction enzyme digestion. Forty-seven children were examined for dental caries and 128 S. mutans clinical isolates cultured from samples of their saliva and plaque and from the saliva of their mothers. Thirty-three children had active caries (70%) and the remainder (n = 14) were caries-free. Caries prevalence was significantly different (p = 0.02) between black African and coloured children, but no differences were found between gtfB amplitypes by caries or ethnic grouping. Thirty-four (27%) of the S. mutans clinical isolates investigated did not ferment melibiose. Melibiose-negative phenotypes (n = 10) isolated from four families showed gtfB RFLP patterns identical to each other. Mothers and children harboured between one and three amplitypes. GtfB amplitypes were identical in 17 families (17/47), of which nine only were identical to S. mutans reference strains. The percentage match between S. mutans amplitype from mothers and their children was low (13%) in the caries-free group compared to children with caries (44%). RFLP analysis of the gtfB gene showed the diversity of S. mutans genotypes within two South African populations that were acquired from mothers and other sources.

Child, Preschool↗

Preventive measures in mothers influence the establishment of the bacterium Streptococcus mutans in their infants.

First-time mothers who had a high salivary number of Strep. mutans [greater than or equal to 10(6) colony-forming-units (c.f.u.) per ml] were selected. Every second mother was given a special preventive programme to reduce her salivary level below 3 x 10(5) c.f.u. per ml. Where a reduction of Strep. mutans was achieved in the mother, the establishment of Strep. mutans in her infant was prevented or delayed. Thus, 28 mothers were successfully treated until their infants were 23 months old and only 3 of their infants (11 per cent) were infected with Strep. mutans, compared with 17 out of 38 infants in the control group (45 per cent). In both groups, the percentage of infected infants increased with increasing age, although at all ages fewer infants were infected with Strep. mutans in the test group than in the control group. Sixteen infants of successfully treated mothers had reached the age of 36 months. Three were infected (19 per cent) compared with 17 out of 27 in the control group (63 per cent). These findings show that the spread of Strep. mutans can be delayed or prevented by measures directed against the main source of infection, an approach which is successful in the prevention of other infectious diseases.

Adult↗

The relationship between the bacterium Streptococcus mutans in the saliva and dental caries in children in Mozambique.

Four hundred and sixty-two children, aged 10-14 years, from eight areas were examined for Streptococcus mutans in saliva and for dental caries. Strep. mutans strains from some samples were further characterized by serological and biochemical methods. Strep. mutans was identified in 98 per cent of the children and 40 per cent showed high counts. Serotypes c and d dominated among the strains. The distribution of Strep. mutans among the children varied between the eight areas but was not correlated with the difference in caries prevalence which ranged from 38 to 88 per cent. In individual children, however, there was an association between high counts of Strep. mutans and the presence of dental caries. Thus, Strep. mutans can be widely distributed and occurs in high numbers in a population with a low prevalence of caries.

Adolescent↗

Effect of monoclonal antibodies against lipoteichoic acid from the oral bacterium Streptococcus mutans on its adhesion and plaque-accumulation in vitro.

Five monoclonal antibodies directed against Streptococcus mutans strain JBP lipoteichoic acid (LTA) were characterized. They were all similarly reactive with the immunizing LTA-containing extract, with intact Strep. mutans JBP cells and with LTA purified from Lactobacillus casei. Immobilized anti-LTA antibodies removes LTA from LTA-containing extracts. The binding of antibodies to LTA was inhibited by the aqueous extract but not by the organic extract of de-acylated LTA, indicating reactivity with the polyglycerol-phosphate portion of the molecule. Antibodies were reactive with all serotypes of Strep. mutans, as well as with strains of Streptococcus salivarius, Streptococcus sanguis and L. casei, but not with LTA-negative species Streptococcus mitis or Actinomyces viscosus. Anti-LTA antibodies at doses of 0.3 or 3.0 micrograms/ml, had no effect on the adherence of Strep. mutans JBP to experimental salivary pellicles formed on hydroxyapatite, but enhanced adherence 150-300 per cent at 30 micrograms/ml. There was no effect of anti-LTA antibodies in a chemostat model which measured sucrose-dependent plaque accumulation by Strep. mutans. The results argue against a major role for LTA in Strep. mutans adherence or plaque accumulation in vitro.

Adhesiveness↗

The influence of Streptococcus mutans on adhesion of Candida albicans to acrylic surfaces in vitro.

Adhesion of Candida albicans and Streptococcus mutans was studied by incubation of radiolabelled cells with acrylic test specimens in a chemically defined growth medium. Strep. mutans adhered firmly in the presence of sucrose, while C. albicans was only loosely attached to the acrylic in both glucose and sucrose media. Firm adhesion of C. albicans occurred when the yeast was incubated simultaneously with Strep. mutans, in the presence of sucrose. The adhesion of C. albicans was also stimulated by incubation with Strep. mutans culture supernatants. Adhesion was not affected by the presence of partially purified glucosyltransferase from Strep. mutans IB. Coaggregation between C. albicans and Strep. mutans upon growth in sucrose medium was observed by light and scanning electron microscopy. No coaggregation was observed in the presence of glucose.

Bacterial Adhesion↗

Agglutination of Streptococcus mutans serotype C cells but inhibition of Porphyromonas gingivalis autoaggregation by human lactoferrin.

The ability of various forms of human lactoferrin (LF) to agglutinate oral Streptococcus mutans, Strep. sobrinus, Strep. rattus, Strep. sanguis, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans cells was studied spectrophotometrically. Fe3+ saturated LF was unable to agglutinate these bacteria, whereas iron-free LF (apo LF) effectively agglutinated Strep. mutans cells but not the other bacteria. The efficiency and rate of agglutination of Strep. mutans were somewhat lower with apo LF than with human whole saliva. However, secretory IgA, phosphate and whole saliva almost totally abolished the apo LF-mediated agglutination of Strep. mutans, suggesting binding to the same target sites on bacterial cell surfaces, or to each other. The presence of exogenous iron (Fe2+, Fe3+), lactoperoxidase or serum albumin did not affect the agglutination by apo LF. Low Ca2+ (50-100 microns) slightly enhanced the agglutination by apo LF but higher concentrations (0.5-1.0 mM) totally blocked the apo LF-mediated agglutination of Strep. mutans. Both saliva and apo LF significantly delayed the rapid autoaggregation of P. gingivalis cells. Aggregation of P. gingivalis is considered a potential virulence factor and a protective mechanism against the host's cellular defences in the gingival crevice. These findings show a novel, strain-specific antibacterial mechanism for LF against Strep. mutans and P. gingivalis and adds a new compound to the group of agglutinating proteins in human saliva.

Agglutination↗

Comparison of Streptococcus mutans and Streptococcus sanguis receptors for human salivary agglutinin.

Oral streptococci vary in their susceptibility to salivary agglutinin-mediated aggregation. To understand the molecular basis of this specificity, the structure and function of receptors for agglutinin from Streptococcus mutans KPSK2 (MSL-1) and Streptococcus sanguis M5 (SSP-5) were compared. Immunological screening of an S. mutans KPSK2 genomic DNA library yielded two identical clones expressing a streptococcal protein that co-migrated with a 220 kDa peptide in SDS extracts from this organism. This protein inhibited agglutinin-mediated aggregation of S. mutans KPSK2 in a dose-dependent manner. The MSL-1 gene is homologous to the S. mutans SpaP and pac genes although single base substitutions alter several amino acids. MSL-1 is also similar to the agglutinin receptor (SSP-5) cloned from S. sanguis M5. All three proteins, MSL-1, P1, and SSP-5 share at least one epitope since monoclonal and polyclonal anti-SSP-5 antibodies react with both MSL-1 and P1. However, other monoclonal antibodies are specific for SSP-5 and appear to react with a peptide domain exhibiting little homology to MSL-1 or P1. Sugar inhibition studies showed that agglutinin-mediated aggregation of S. mutans KPSK2 was most potently inhibited by fucose and lactose. Sialic acid, a potent inhibitor of S. sanguis aggregation, had no effect on the interaction of agglutinin with S. mutans KPSK2. These results suggest that while the MSL-1 and SSP-5 proteins are genetically and immunologically related, their specificity for binding sites on agglutinin differs.

Adhesins, Bacterial↗

Contribution of cell surface protein antigen PAc of Streptococcus mutans to bacteremia.

Streptococcus mutans, a major cariogenic bacterium, is occasionally isolated from the blood of patients with bacteremia and infective endocarditis. Mutant strains of S. mutans MT8148, defective in the major surface proteins glucosyltransferase (GTF) B-, C-, and D-, and protein antigen c (PAc), were constructed by insertional inactivation of each respective gene with an antibiotic resistant cassette. Susceptibility to phagocytosis was determined by analyses of interactions of the bacteria with human polymorphonuclear leukocytes, and the PAc-defective mutant strain (PD) showed the lowest rate of phagocytosis. Further, when PD and MT8148 were separately injected into the jugular veins of Sprague-Dawley rats, PD was recovered in significantly larger numbers and for a longer duration, and caused more severe systemic inflammation than MT8148, indicating that S. mutans PAc is associated with its systemic virulence in blood. Next, 100 S. mutans clinical isolates from 100 Japanese children and adolescents were analyzed by Western blotting using antisera raised against recombinant PAc, generated based on the pac sequence of MT8148. Four of the 100 strains showed no positive band and each exhibited a significantly lower phagocytosis rate than that of 25 randomly selected clinical strains (P < 0.01). In addition, three of the 100 strains possessed a lower molecular weight PAc and a significantly lower rate of phagocytosis than the 25 reference strains (P < 0.05). These results suggest that S. mutans PAc may be associated with phagocytosis susceptibility to human polymorphonuclear leukocytes, with approximately 7% of S. mutans clinical isolates possible high-risk strains for the development of bacteremia.

Animals↗

Colonization and vertical transmission of Streptococcus mutans in Turkish children.

The aim of the study was to establish the colonization of Streptococcus mutans and to determine the possibility of intra-familial transmission in a group of Turkish children and their parents. A total of 56 children participated in the study together with their parents (20 fathers and 49 mothers). Saliva samples were collected from the individuals and cultivated on S. mutans selective TYCSB agar. The typical isolates of S. mutans were identified by using classical microbiological methods, as well as molecular typing of S. mutans clones which was performed by using AP PCR with OPA5 primer for the detection of transmission. The vertical transmission of salivary S. mutans was detected among 14 mother-father-child, 35 mother-child (one twins) and 6 father-child combinations. The homologies of strain types were recorded as 24% and 16.6% for mother-child and father-child combinations, respectively. A significant positive correlation (p<0.001) was found between the infected children and their parents with high S. mutans counts.

Adult↗

Relationships between dietary behaviours, oral hygiene and mutans streptococci in dental plaque of a group of infants in southern England.

This report is part of a prospective study on dietary behaviours, feeding practices, oral hygiene and dental health during the first 18 months of life in a cohort of infants living in southern England. In this part of the study the prevalence of mutans streptococci in the plaque of 1-year-old children and its correlation with sociodemographic status, dietary behaviour and oral hygiene practices over the first 12 months of life were studied. Dietary habits, oral hygiene and dental health at 18 months were also studied. The study group comprised 163 infants. At 12 months of age all children had a dental examination and a plaque sample was removed from the labial surfaces of upper incisors for microbiological examination. A further dental examination took place at 18 months of age; no plaque was sampled. Longitudinal dietary information was obtained with a 3-day food diary at 6, 12 and 18 months of age. Parents/carers completed two structured questionnaires on sociodemographic factors, oral hygiene and feeding behaviour over the first 18 months of life when their children were 12- and 18-month-old. No child exhibited dental caries at 12 and 18 months of age; 18 and 25% had visible dental plaque at 12 and 18 months, respectively. Seven children (4%) had detectable mutans streptococci in their plaque at 12 months of age. Children who had started brushing their teeth or who had had their teeth brushed by 12 months of age were less likely to have detectable mutans streptococci in their plaque than those who had not (P=0.02). The amount of mutans streptococci was significantly correlated with the total number of eating/drinking events per day (P<0.001) and bordered on significant correlation with the mean daily frequency of consumption of foods and drinks containing non-milk extrinsic sugars (P=0.05). None of the sociodemographic variables-sex, social class, mother's level of education-were associated with the detection of mutans streptococci in this group of infants. Multiple logistic regression analysis revealed that two variables were independently and positively correlated with the presence of mutans streptococci in the plaque; these were age when tooth brushing started and total eating and drinking events per day.

Age Factors↗

Characterization of the mucosal immune response in breast milk after peroral immunization of chimpanzees (Pan troglodytes) with Streptococcus mutans.

The characteristics of the mucosal immune response to Streptococcus mutans cells, antigen A, antigen B, glucosyltransferases and glucan-binding proteins were examined in four pregnant chimpanzees that had been immunized perorally with Strep. mutans. Six pregnant chimpanzees served as non-immunized controls. None of the chimpanzees harbored S. mutans. Samples of milk were collected from all animals throughout the experiment. Peroral immunization resulted in an overall 17-fold median increase in SIgA in milk. Although SIgA1 comprised almost two-thirds of milk SIgA, Strep. mutans whole-cell antibody activity was contained predominantly in the SIgA2 subclass. The difference between the specific activities of anti-Strep. mutans SIgA1 and SIgA2 antibodies compared over time reached the borderline of statistical significance (p = 0.08). The avidity of anti-Strep. mutans antibodies was low in three of four chimpanzees and there was no evidence of affinity maturation. SIgA antibodies from the milk of all four immunized chimpanzees recognized antigen A. In three animals these antibodies were restricted to the SIgA1 subclass and, in one animal, anti-A antibodies were confined to SIgA2. Antibodies from all of the immunized chimpanzees recognized degradation products of antigen B in both the SIgA1 and the SIgA2 subclasses. Only two of four immunized chimpanzees responded to glucosyltransferases and these antibodies were restricted to the SIgA1 subclass. None of the chimpanzees responded to the 74-kDa glucan-binding protein. However, three animals produced SIgA1 antibodies against the 59-kDa glucan-binding protein and two of these also produced SIgA2 antibodies against this protein.

Analysis of Variance↗

Effect of water storage of E-glass fiber-reinforced composite on adhesion of Streptococcus mutans.

This study investigated the effect of water storage of fiber-reinforced composite on the adhesion of Streptococcus mutans (S. mutans) and its ability to stay adhered and multiply on the FRC. The materials (E-glass fibers and denture base polymer) were stored in water for 14 or 30 days or left dry. Water contact angles of the materials before and after water storage were determined. Test specimens, with or without parotid saliva or serum pellicle, were incubated in a suspension of S. mutans allowing initial adhesion to occur. Bacterial adhesion and multiplication was studied using scanning electron microscopy. Contact angles of both materials were significantly reduced after water storage indicating an increase in surface free energy. When studied without a surface pellicle, water storage significantly increased adhesion of S. mutans to both glass and polymer. Saliva coating of the materials resulted in higher degree of adhesion to glass fibers in comparison with polymer and after 14 days water storage glass bound over twice as much S. mutans cells than the polymer matrix. Bacterial growth and biofilm formation occurred equally on both materials. The results of this in vitro study suggest that in order to avoid the possible increase in S. mutans adhesion, the reinforcing glass fibers should be covered with the matrix polymer of the composite.

Bacterial Adhesion↗