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PubMedDate not supplied

Inhibitory effects of MoAbs against a surface protein antigen in real-time adherence in vitro and recolonization in vivo of Streptococcus mutans.

A surface protein antigen (PAc) of Streptococcus mutans, particularly the A-region of the molecule, has been reported to interact with salivary components on the tooth surface. It might be a candidate antigen inducing the production of antibodies against the adherence of S. mutans to the tooth surface. We investigated the effects of monoclonal antibodies (MoAbs) obtained by immunization of synthetic PAc peptides that completely correspond to the amino acid sequence of part of the A-region. These MoAbs recognize several core B-cell epitopes in the sequence. Two (KH5 and SH2) of these antibodies reacted with both S. mutans and Streptococcus sobrinus, but not with Streptococcus sanguis, Streptococcus salivarius, Porphyromonas gingivalis or Lactobacillus casei. They clearly inhibited the real-time adherence of S. mutans to salivary components in a biosensor. KH5, which showed a real-time inhibition (71%), also significantly prevented the recolonization of S. mutans on the tooth surface in rats. These results suggested that the core B-cell epitope (-Y---L--Y----) recognized by KH5 was the essential sequence in the antigenic epitopes of PAc protein recognized specifically by the inhibitory antibody. Therefore, the amino acid residues were found to be important in the initial attachment of S. mutans to the tooth surface. These results provide for the mechanism of PAc molecule in the initial attachment of S. mutans on the tooth surface and more effective designs for the removal of S. mutans and S. sobrinus from the oral cavity.

Adhesins, Bacterial↗

PubMed2006

In vitro binding interactions of oral bacteria with immobilized fructosyltransferase.

AIMS: The objective of the present study was to explore the role of immobilized fructosyltransferase (FTF) in adhesion process. METHODS AND RESULTS: We investigated real-time biospecific interactions between several types of oral bacteria and recombinant FTF immobilized on a biosensor chip, using surface plasmon resonance technology. Streptococcus mutans, Streptococcus sobrinus and Actinomyces viscosus demonstrated significant binding to FTF. Actinomyces viscosus had a greater binding to FTF, with 373 Resonance Units (RU), than the other tested bacteria. The binding level to FTF of Strep. sobrinus was 320 RU, whereas Strep. mutans and Streptococcus salivarious show binding of 296 and 245 RU, respectively. The binding sensograms displayed different profiles for the tested bacteria at various cell density, suggesting a different affinity to immobilized FTF. CONCLUSIONS: The results from this study suggest that FTF may influence bacterial adherence and colonization of the dental biofilm. SIGNIFICANCE AND IMPACT OF THE STUDY: The biomolecular interaction analysis enables real-time monitoring of the interaction between adhesions of intact bacteria and their ligands, which might be crucial in the initial phase of biofilm development in vivo.

Actinomyces viscosus↗

PubMed2001

Cloning and DNA sequencing of the surface protein antigen I/II (PAa) of Streptococcus cricetus.

We have cloned and sequenced the gene encoding the surface protein antigen PAa (antigen I/II family) from Streptococcus cricetus E49 (serotype a) using degenerate PCR. The deduced amino acid sequence of PAa reveals two repeating regions (A region; alanine-rich region, P region; proline-rich region). Two additional tandem repeats were found in the A region and part of the P region was deleted compared to antigen I/II. Homology and phylogenetic analyses reveal that PAa is homologous to Streptococcus sobrinus PAg rather than Streptococcus mutans PAc. Using degenerate PCR a gene homologous to PAc was identified in Streptococcus intermedius, but not found in Streptococcus rattus or Streptococcus anginosus.

Animals↗

PubMed1989

Plaque inhibition by a combination of zinc citrate and sodium lauryl sulfate.

Bacteriological tests demonstrated an additive inhibitory effect of ZnCl2 and sodium lauryl sulfate (SLS) on in vitro growth of Streptococcus sobrinus OMZ 176 and of Streptococcus sanguis ATCC 10556. As measured by atomic absorption spectrophotometry, the solubility of zinc citrate increased in the presence of SLS. After 48 h, the concentration of solubilized zinc from aqueous solutions of 5.0 mM zinc citrate was 12.0 mM versus 14.4 mM in the presence of 34.7 mM SLS. The plaque-inhibiting properties of aqueous solutions of 12.0 mM Zn2+ from zinc citrate, 34.7 mM SLS, and 14.4 mM Zn2+ from zinc citrate in combination with 34.7 mM SLS were examined in 7 volunteers. Plaque accumulations were assessed by using a method earlier described after 3 days of twice daily mouthrinses with 10 ml test solution, during which period no mechanical oral hygiene was performed. Compared to placebo, SLS and zinc citrate increased the frequency of plaque index score 0 by 52.9 and 98.3%, respectively, and SLS gave 36.9% and zinc citrate 55.7% less surfaces with scores 2 or 3 (p less than 0.05 in all cases). The combination of zinc citrate and SLS gave a threefold increase of plaque index score 0 and a 70.5% reduction of scores 2 or 3 (p less than 0.05).

Chemical Phenomena↗

PubMed1993

Phenotypic characterization of oral streptococci by classical methods.

The phenotypic characteristics of 215 strains of oral streptococci were compared on the basis of the results of sorbitol, raffinose and trehalose fermentation, ammonium from arginine production, aesculin hydrolysis, H2O2 production, susceptibility to 2 U bacitracin, and growth in 4% NaCl solution. Eleven different species were identified. Among the most noteworthy findings were the low number of H2O2-producing strains of Streptococcus sobrinus, the effective discrimination between Streptococcus sanguis and Streptococcus mitis on the basis of raffinose fermentation, and the phenotypic similarity between Streptococcus oralis and the former mutans group. To avoid the creation of new species, the term biotype is proposed to designate phenotypically distinct organisms which do not qualify as different species.

Dental Plaque↗

PubMedDate not supplied

Differences in acidogenicity of S. sobrinus and S. rattus are linked to the catalytic efficiency of the glycolytic key enzyme phosphofructokinase.

This contribution describes the biochemical properties of two catalytically different phosphofructokinases (PFKs) purified from Streptococcus rattus LB 2 (PFK-rat) and Streptococcus sobrinus OMZ 65 (PFK-sob), respectively. Steady-state kinetics revealed K(M) = 0. 8 mM for PFK-rat and K(M) = 0.08 mM for PFK-sob for F-6-P as the substrate. The enzymes also differ in their pH profiles: whereas the highest activity of PFK-rat was measured at pH = 8.0, the optimum pH of PFK-sob was at pH = 7.0. In addition, compared to PFK-sob, PFK-rat was more sensitive against the allosteric inhibitor ATP. PFK catalyzes a committed step of glycolysis, the main acid producing catabolic pathway. Thus, the catalytically more efficient enzyme isolated from S. sobrinus OMZ 65, especially at low pH, could explain the comparably high acidogenicity of this strain.

Bacterial Proteins↗

PubMed1996

The inhibitory action of fatty acids on oral bacteria.

Saturated and unsaturated fatty acids and fatty acid derivatives were examined for their growth-inhibitory effects towards three selected oral bacteria: Porphyromonas gingivalis, Selenomonas artemidis, and Streptococcus sobrinus. Of the 45 compounds surveyed, only one, myristoleic acid, was inhibitory towards S. artemidis at a concentration < 100 micrograms/ml. cis-Hexadecenoic and cis-octadecenoic acids were generally inhibitory towards P. gingivalis and S. sobrinus, but there was no correlation between the position of the double bond and the minimum inhibitory concentration. Supra-minimum inhibitory concentrations of palmitoleic acid did not promote leakage of intracellular materials from either P. gingivalis or S. sobrinus, nor was L-isoleucine uptake by S. sobrinus inhibited. Fatty acids and derivatives were also examined for prospective synergistic or antagonistic interactions with thymol vis-à-vis growth inhibition of the test strains. Lauric acid and myristic acid each behaved synergistically with thymol to inhibit the growth of at least one test strain, whereas cis-10-heptadecenoic acid and thymol were noticeably antagonistic towards the growth of S. sobrinus.

Bacteria, Anaerobic↗

PubMed1994

Localization on the cell surface of streptococcal protein antigen A.

The cellular localization of the major surface protein SpaA of Streptococcus sobrinus 6715 was examined by immunoelectron microscopy with rabbit polyclonal antibodies directed against purified SpaA protein. Immunoelectron microscopic analysis of thin sections of S. sobrinus cells revealed that the SpaA protein is associated with the fibrillar fuzzy coat of S. sobrinus cells and appears to be distributed over the entire surface of S. sobrinus cells.

Antigens, Bacterial↗

PubMed1991

In vitro demineralization of enamel by F-sensitive and F-resistant mutans streptococci in the presence of 0, 0.05, or 0.5 mmol/L NaF.

Lactate production and accompanying enamel demineralization by fluoride-sensitive and fluoride-resistant mutans streptococci were studied in an in vitro demineralization model in the presence of 0, 0.05, or 0.5 mmol/L NaF. The fluoride-resistant strains were derived from laboratory strains or were recently isolated strains from xerostomic patients on high-dose fluoride therapy. The demineralization model was composed of a cell suspension in a glucose-agarose gel overlying a bovine enamel block. Lactate and calcium content of the agarose were determined after 22-hour incubations at 37 degrees C. Fluoride-resistant variants of Streptococcus sobrinus 6715-15 produced less lactate and caused less demineralization than did the parent strain even in the presence of fluoride. On the other hand, fluoride-resistant variants of Streptococcus mutans C180-2 and of S. mutans GS-5 produced more acid and caused greater demineralization than did their respective parent strains, both in the absence and presence of fluoride. Two recently isolated fluoride-resistant S. mutans strains produced more lactate and demineralized enamel more than did two recently isolated S. mutans strains from normal human subjects, both in the presence of 0 and 0.05 mmol/L NaF. It is concluded that adaptation to fluoride resistance does not invariably reduce the cariogenicity of mutans streptococci nor the effectiveness of fluoride in preventing demineralization.

Animals↗

PubMed2004

Effect of a high-molecular-weight component of cranberry on constituents of dental biofilm.

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.

Bacterial Adhesion↗

PubMed1991

[Caries-inducing activity of soybean-oligosaccharide (SOR) in vitro and in experimental dental caries of rats].

The caries-inducing activity of soybean-oligosaccharide (SOR: stachyose- and raffinose-rich sugar mixture) was examined in in vitro and in vivo experiments. Streptococcus mutans MT 8148R fermented SOR and produced acids. However Streptococcus sobrinus 6715 did not ferment. SOR was not able to act as a substrate for crude glucosyltransferases (GTase) of these mutans streptococci to synthesize the water-insoluble glucan. However, SOR did not inhibit the synthesis of water-insoluble glucan from sucrose by crude GTase. SOR was proved to be of low cariogenicity in rats infected with S. sobrinus 6715.

Animals↗

PubMed2001

[The isolation and identification of pathogenic bacteria from rampant caries in children].

OBJECTIVE: In order to prevent rampant caries in children, the predominant pathogenic bacteria of rampant caries in children were defined. METHODS: The pathogenic bacteria of rampant caries were isolated and identified in 30 children with rampant caries, aged from 2 to 5 years old. Samples were collected from the caries of labial surfaces of maxillary anterior deciduous teeth, including caries lesions and caries-free smooth enamel surfaces. Plaque were obtained from the cervical third areas of the maxillary labial anterior deciduous tooth surfaces in children without rampant caries and caries-free children and served as controls respectively. The isolates were identified by colony morphology, biochemical tests and DNA base contents (mol percent G + C). RESULTS: The prevalence of Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus) in the plaque of caries lesions and the proportions of S. mutans and S. sobrinus in the plaque of 2 sampling sites of children with rampant caries were all significantly higher than those of children without rampant caries and caries-free children (P < 0.05). CONCLUSION: S. mutans and S. sobrinus are predominant pathogenic bacteria in rampant caries in children.

Child↗

PubMed1995

Effect of aluminum concentration on dental caries formation in the rat.

OBJECTIVE: Topically applied aluminum (AI) reduced fissure caries in rats with indigenous microflora, but neither therapeutic activity nor concentration effects have been studied in animals infected with caries-inducing bacteria. This study investigated the effects of solutions containing various AI concentrations on formation of smooth surface and sulcal caries in cariogenically challenged rats infected with Streptococcus sobrinus. METHODS: Weanling rats, inoculated with S. sobrinus, were randomly assigned to six groups which were topically treated twice daily with water (treatment control), a NaF solution of 100 parts/10(6) F (positive control), or AIK(SO4)2 solutions containing 100, 1000, 2000, or 4000 parts/10(6) AI. Rats were fed a high-sucrose diet and distilled water ad libitum for 10 weeks, then the molars were scored for enamel and dentin caries. RESULTS: Relative to the water control, NaF and all AI solutions significantly reduced prevalence of both smooth-surface and fissure caries. Protection by AI solutions was greater on smooth surfaces than sulcal areas. AI treatments reduced smooth surface scores by 49-71% and sulcal scores by 27-53%. For all comparisons the protective effect of the 100 parts/10(6) AI solution was less than solutions containing 1000 parts/10(6) AI or more. NaF and all AI solutions also significantly reduced the extent of lesions penetration into dentin. CONCLUSIONS: Topically applied AI reduced the formation and progression of both smooth surface and sulcal caries and showed evidence of a dose response in a rat model infected with S. sobrinus.

Administration, Topical↗

PubMed1992

Synergistic degradation of bovine serum albumin by mutans streptococci and other dental plaque bacteria.

Mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) exhibited low levels of proteolytic activity against the model protein substrate, FITC-labelled bovine serum albumin, when incubated alone. Inclusion of other members of the dental plaque microflora in the assay usually resulted in marked increases in the degree of proteolysis and a high level of synergy. Interactions between mutans streptococci and either Streptococcus oralis or Fusobacterium nucleatum gave rise to the greatest degree of synergistic proteolytic degradation.

Dental Plaque↗

PubMed1997

Immunoglobulin A reaction to oral streptococci in saliva of subjects with different combinations of caries and levels of mutans streptococci.

The aim of this study, performed in Bangkok, was to study whether a particular salivary immunoglobulin A (IgA) antibody profile against mutans streptococci could be related to the absence or presence of caries. A group of 12-year-old individuals representing various combinations of mutans streptococci levels and caries experience was selected. Whole saliva stimulated by paraffin-chewing was collected, and the children were investigated for decayed, missing and filled surfaces (DMFS) and teeth (DMFT), following WHO criteria and methods, at baseline and after 2 years. The total amount of salivary IgA was determined by an immunobead enzyme-linked immunosorbent assay, and SDS-PAGE and Western blot analysis was performed using sonicated antigens of Streptococcus mutans and Streptococcus sobrinus strains and, as a control, a Streptococcus parasanguis strain. The results showed that Thai children with low caries prevalence had more distinct immunoblot bands to antigens from mutans streptococci than did the high-caries children. A similar picture was not seen for S. parasanguis. On the whole, the Thai children also showed fewer bands than usual Swedish saliva samples from comparable age groups. The complexity of the relationship between dental caries and IgA in saliva is highlighted.

Adolescent↗

PubMed1987

Characterization of a rat salivary sialoglycoprotein complex which agglutinates Streptococcus mutans.

Rat saliva agglutinated Streptococcus mutans Ingbritt and NCTC 10449 and Streptococcus sanguis NCTC 7864 but not S. mutans NCTC 10921, GS 5, or LM 7, Streptococcus sobrinus 6715-13 or OMZ 65, or Streptococcus cricetus HS 6, as measured turbidometrically. The specificity of agglutination by rat saliva was the same as that by human saliva. Agglutination was associated with a mucin complex (rat salivary agglutinin complex [rS-A]) of sulfated sialoglycoproteins, with a trace of associated lipid and an apparent Mr of 1.6 X 10(6), isolated by gel-filtration Fast Protein Liquid Chromatography. The complex was dissociated in a high-ionic-strength buffer containing 6 M urea and then fractionated by gel filtration and anion-exchange Fast Protein Liquid Chromatography into four sulfated sialoglycoprotein components, designated rS-A-1Q1, rS-A-1Q2, rS-A-1Q3, and rS-A-2, with rS-A-1Q2 being polydisperse through differential glycosylation of the polypeptide backbone. The dissociation destroyed agglutination activity. The polypeptide backbones contained up to 42% serine plus threonine and up to 40% glycine plus alanine plus proline plus valine. The carbohydrate moiety of the rS-A sialoglycoproteins consisted of N-acetylgalactosamine, sialate, galactose, fucose, N-acetylglucosamine, and small amounts of mannose, with the predominant sugar being N-acetylgalactosamine. Agglutination was inhibited by 1 mM EDTA but was restored by 1.5 mM CaCl2. Agglutination was also inhibited by 5 mM CaCl2; nonimmune sera; cationic polymers; and wheat germ, lentil, soybean, and peanut lectins. However, agglutination was not affected by lipoteichoic acid, various anionic proteins, or various sugars. Neuraminidase treatment of rS-A did not affect activity, but tryptic digestion of S. mutans did prevent agglutination. The results are consistent with calcium bridging the negative groups within the rS-A complex and allowing the approach of rS-A to the bacterial cell surface to effect a specific conformational attachment.

Agglutinins↗

PubMedDate not supplied

Antimicrobial effect of a novel ozone- generating device on micro-organisms associated with primary root carious lesions in vitro.

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.

Anaerobiosis↗

PubMed2004

Detection of cariogenic bacterial genes by microchip electrophoresis.

Allele-specific PCR primers were designed, based on the dextranase (dex) gene, to identify Streptococcus mutans and Streptococcus sobrinus in dental plaque; subsequently, PCR products were detected via microchip electrophoresis (ME). In order to amplify the dex gene fragment of S. mutans and S. sobrinus, the following two PCR methods were established. Duplex allele-specific PCR primers were designed on a region of low DNA homology; furthermore, 211 and 126-bp fragments were amplified for S. mutans and S. sobrinus, respectively. Common PCR primer for single allele-specific PCR was designed so as to sandwich a region exhibiting high homology and amplify PCR product of different DNA size due to deletion of small DNA fragment in two dex genes. S. mutans and S. sobrinus were amplified, leading to the generation of 202 and 226-bp products, respectively. Analysis of DNA base size by ME in order to achieve efficient separation employed a polymer mixture consisting of hydroxypropyl methylcellulose (HPMC) and polyethylene oxide (PEO). In the presence of a polymer mixture of 0.125% PEO/0.6% HPMC, two PCR products were obtained, displaying degree of separation of 226 bp/202 bp of 2.67 (Rs). Reproducibility (CV%, n = 7) was 0.3%; additionally, separation time was approximately 85 s. This method was applied to the detection of S. mutans and S. sobrinus in dental plaque. Detection of the dex genes of S. mutans and S. sobrinus characterized by quickness, precision and high sensitivity was possible.

Alleles↗