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1.
J Appl Microbiol ; 126(3): 740-751, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30556937

RESUMO

AIMS: To investigate the effects of the ginsenoside Rh2 on monospecies and multispecies cariogenic biofilms and explore the mechanism of the antibiofilm effect of Rh2 in vitro. METHODS AND RESULTS: Streptococcus mutans, Streptococcus sobrinus and Streptococcus sanguinis were chosen to form the monospecies or multispecies biofilms. Crystal violet staining and laser scanning confocal microscopy were used to observe the effect of Rh2 on biofilms in vitro. Cytotoxicity was examined by the Cell Counting Kit-8. The effects of Rh2 on bacterial membranes were observed via transmission electron microscopy (TEM). The isobaric tags for relative and absolute quantification (iTRAQ) method were used to profile the common differentially expressed proteins. Gene expression was analysed by reverse transcription quantitative polymerase chain reaction. In general, the treatment of cariogenic biofilms with Rh2 significantly decreased biomass accumulation by inhibiting bacterial growth and extracellular polysaccharide synthesis without any cytotoxic effects. TEM imaging showed that Rh2 could disrupt the cell membranes of these bacteria. The iTRAQ results indicated that the levels of mannose-specific IIC/D and acetaldehyde/alcohol dehydrogenase were substantially down-regulated, while the mRNA expression of the corresponding genes were significantly changed. CONCLUSIONS: Our data revealed a potential application for Rh2 in the protection against dental caries via the inhibition of cariogenic biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes the first application of a ginsenoside against multispecies cariogenic biofilms. Rh2 may serve as an alternative agent to prevent dental caries by effectively modulating the pathogenic potentials of oral biofilms.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Ginsenosídeos/farmacologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus sanguis/efeitos dos fármacos , Streptococcus sobrinus/efeitos dos fármacos , Cárie Dentária/microbiologia , Humanos , Streptococcus mutans/genética , Streptococcus mutans/fisiologia , Streptococcus sanguis/genética , Streptococcus sanguis/fisiologia , Streptococcus sobrinus/genética , Streptococcus sobrinus/fisiologia
2.
Mar Drugs ; 16(9)2018 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-30150576

RESUMO

Dental plaque biofilms cause various dental diseases; therefore, inhibiting the growths of the dental plaque bacteria which produce biofilms can be a strategy for preventing dental disease. Certain sulfated polysaccharides from marine algae exert antimicrobial activities against human bacterial pathogens in addition to their physiological benefits. On the basis of these observations, the antimicrobial and antibiofilm activities of sulfated polysaccharides from different marine algae were evaluated against dental plaque bacteria. Among the sulfated polysaccharides, a fucoidan from Fucus vesiculosus showed notable antimicrobial activities against the selected dental plaque bacteria, including some foodborne pathogenic bacteria. The minimum inhibitory concentrations were of 125 to 1000 µg mL-1. Regarding the antibiofilm activity, the fucoidan at the concentrations of above 250 µg mL-1 completely suppressed the biofilm formations and planktonic cell growths of Streptococcus mutans and S. sobrinus. However, no eliminative effect on the completed biofilm was observed. The fucoidan consisted of almost fucose base polysaccharide containing approximately 14.0% sulfate content. The average molecular weight of the fucoidan was changed by heat treatment (121 °C for 15 min) and it affected the antimicrobial activity.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Placa Dentária/tratamento farmacológico , Fucus/química , Polissacarídeos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antibacterianos/uso terapêutico , Organismos Aquáticos/química , Placa Dentária/microbiologia , Avaliação Pré-Clínica de Medicamentos , Humanos , Testes de Sensibilidade Microbiana , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/uso terapêutico , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologia , Streptococcus sobrinus/efeitos dos fármacos , Streptococcus sobrinus/fisiologia , Ésteres do Ácido Sulfúrico/química
3.
Anaerobe ; 35(Pt A): 54-9, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25805186

RESUMO

The aim of this pilot study was to assess the difference in virulence of acidogenic and aciduric oral streptococci in an in vitro caries model using their penetration depths into dental enamel. 30 caries-free extracted molars from 11- to 16-year-olds were cleaned ultrasonically for 1 min with de-ionized water and, after air-drying, embedded in epoxy resin. After 8-h of setting at room temperature, the specimens were ground on the buccal side with SiC-paper 1200 (particle size 13-16 µm). Enamel was removed in circular areas sized 3 mm in diameter; the mean depth of removed enamel was 230 ± 60 µm. 15 specimens each were incubated anaerobically under standardized conditions with 24 h-cultures of Streptococcus sanguinis 9S or Streptococcus sobrinus OMZ 176 in Balmelli broth at 37 ± 2 °C; the pH-values of the broths were measured at the beginning and end of each incubation cycle. After 2, 4, 6, 8, and 10 weeks 3 teeth each were fixed in 2.5% glutaraldehyde in cacodylate buffer for 24 h, washed 3× and dehydrated 30-60min by sequential washes through a series of 30-100% graded ethanol. The teeth were cut in half longitudinally; afterward, two slits were made to obtain fracture surfaces in the infected area. After critical-point-drying the fragments were gold-sputtered and viewed in a scanning electron microscope at magnifications of ×20-20,000. After 10 weeks of incubation, penetration of S. sanguinis of 11.13 ± 24.04 µm below the break edges into the enamel was observed. The invasion of S. sobrinus reached depths of 87.53 ± 76.34 µm. The difference was statistically significant (paired t test: p = 0.033). The experimental penetration depths emphasize the importance of S. sanguinis versus S. sobrinus in the context of the extended ecological plaque hypothesis.


Assuntos
Cárie Dentária/microbiologia , Esmalte Dentário/microbiologia , Streptococcus sobrinus/fisiologia , Streptococcus/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Humanos , Projetos Piloto , Streptococcus/genética , Streptococcus sobrinus/genética
4.
Eur J Orthod ; 37(6): 565-9, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25564502

RESUMO

OBJECTIVE: To analyze in vivo mutans streptococci (MS) adhesion to self-ligating ceramic brackets [Clarity-SL (CSL) and Clippy-C (CC)] and the relationships between bacterial adhesion and oral hygiene indices. MATERIALS AND METHODS: Four central incisor brackets from the maxilla and mandible were collected from 40 patients (20 patients per each bracket type) at debonding immediately after plaque and gingival indices were measured. Adhesions of Streptococcus mutans, S. sobrinus, and total bacteria were quantitatively determined using real-time polymerase chain reaction after genomic DNA was extracted. Factorial analysis of variance was used to analyze bacterial adhesion to the brackets with respect to the bracket type and jaw position. Correlation coefficients were calculated to determine the relationships of bacterial adhesion to oral hygiene indices. RESULTS: Adhesion of total bacteria and S. mutans to CSL was higher than that to CC (P < 0.001). Adhesion of total bacteria to the mandibular brackets was higher than that to the maxillary ones (P < 0.001), while adhesion of S. mutans to the maxillary brackets were higher than that in the mandibular ones (P < 0.001). In particular, the proportion of S. mutans to total bacteria in CSL was higher than CC (P < 0.05) in the maxillary anterior teeth (P < 0.001). There were no significant differences in adhesion of S. sobrinus between the brackets and jaw positions. Interestingly, no significant relationships were found between bacterial adhesions and oral hygiene indices. LIMITATIONS: Complex bracket configurations may significantly influence bacterial adhesion to orthodontic brackets. Further in vivo study using bracket raw materials will help to define the relationships between bacteria adhesion and enamel demineralization. CONCLUSIONS: Because oral hygiene indices were not significantly correlated with adhesions of MS to self-ligating ceramic brackets, careful examinations around the brackets should be needed to prevent enamel demineralization, regardless of oral hygiene status.


Assuntos
Aderência Bacteriana/fisiologia , Braquetes Ortodônticos/microbiologia , Streptococcus mutans/fisiologia , Adolescente , Carga Bacteriana , Cerâmica/química , DNA Bacteriano/análise , Materiais Dentários/química , Placa Dentária/microbiologia , Índice de Placa Dentária , Feminino , Humanos , Masculino , Mandíbula/microbiologia , Maxila/microbiologia , Índice de Higiene Oral , Índice Periodontal , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Streptococcus mutans/isolamento & purificação , Streptococcus sobrinus/isolamento & purificação , Streptococcus sobrinus/fisiologia , Propriedades de Superfície , Adulto Jovem
5.
J Appl Oral Sci ; 32: e20230458, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38922241

RESUMO

OBJECTIVE: This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models. METHODOLOGY: In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey's post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001). RESULTS: In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols. CONCLUSION: The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth.


Assuntos
Biofilmes , Cárie Dentária , Esmalte Dentário , Lacticaseibacillus casei , Streptococcus mutans , Humanos , Streptococcus mutans/fisiologia , Cárie Dentária/microbiologia , Cárie Dentária/terapia , Esmalte Dentário/microbiologia , Esmalte Dentário/química , Lacticaseibacillus casei/fisiologia , Fatores de Tempo , Reprodutibilidade dos Testes , Streptococcus sobrinus/fisiologia , Análise Espectral Raman , Análise de Variância , Microscopia de Polarização , Estatísticas não Paramétricas , Remineralização Dentária/métodos , Valores de Referência , Saliva/microbiologia , Saliva/química , Desmineralização do Dente/microbiologia , Fluorescência
6.
BMC Microbiol ; 13: 157, 2013 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-23848601

RESUMO

BACKGROUND: Streptococcus mutans and Streptococcus sobrinus are associated with the development of dental caries in humans. However, previous diagnostic systems are unsuitable for monitoring viable cell numbers in oral specimens. Assessing the relationship between the numbers of viable and dead bacterial cells and oral status is important for understanding oral infectious diseases. Propidium monoazide (PMA) has been reported to penetrate dead cells following membrane damage and to cross-link DNA, thereby inhibiting DNA amplification. In the present study, we established an assay for selective analysis of two viable human cariogenic pathogens, S. mutans and S. sobrinus, using PMA combined with real-time PCR (PMA-qPCR). RESULTS: We designed species-specific primer sets for S. mutans and S. sobrinus, generated standard curves for measuring cell numbers, and evaluated the dynamic range of the assay. To determine the effectiveness of the assay, PMA was added to viable and autoclave-killed cell mixtures. PMA treatment effectively prevented DNA amplification from dead cells. No amplification of DNA from dead cells was observed in these organisms. In addition, we applied this assay to analyze viable cell numbers in oral specimens. A significant correlation was found between the number of viable S. mutans cells in saliva and that in plaque among caries-free patients, whereas no correlation was observed between saliva and carious dentin. The total and viable cell numbers in caries-positive saliva were significantly higher than those in caries-free saliva. Finally, we analyzed the usefulness of this assay for in vitro oral biofilm analysis. We applied PMA-qPCR for monitoring viable S. mutans cell numbers in vitro in planktonic cells and oral biofilm treated with hydrogen peroxide (H2O2). In planktonic cells, the number of viable cells decreased significantly with increasing H2O2 concentration, whereas only a small decrease was observed in biofilm cell numbers. CONCLUSIONS: PMA-qPCR is potentially useful for quantifying viable cariogenic pathogens in oral specimens and is applicable to oral biofilm experiments. This assay will help to elucidate the relationship between the number of viable cells in oral specimens and the oral status.


Assuntos
Biofilmes , Viabilidade Microbiana , Boca/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Coloração e Rotulagem/métodos , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Azidas/metabolismo , Carga Bacteriana/métodos , Criança , Pré-Escolar , Primers do DNA/genética , Feminino , Humanos , Masculino , Propídio/análogos & derivados , Propídio/metabolismo , Streptococcus mutans/isolamento & purificação , Streptococcus sobrinus/isolamento & purificação
7.
Anaerobe ; 19: 1-8, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23142795

RESUMO

The aim of this study was to evaluate the effect of the methanol extract of Withania somnifera (MEW) on the growth and virulence properties of Streptococcus mutans and Streptococcus sobrinus at sub-minimum inhibitory concentration (MIC) levels and to identify the main components of MEW. First, antibacterial activity of MEW against oral bacteria was determined using a micro-dilution method. Then, the effect of MEW on the growth of S. mutans and S. sobrinus was investigated at sub-MIC levels. To test the effect of MEW on the virulence properties of S. mutans and S. sobrinus, assays for acid production, acid tolerance, and biofilm formation were performed at sub-MIC levels. A GC-MS analysis for the main components of MEW was also carried out. MEW showed a broad antibacterial range against oral bacteria (MIC: 0.125-2 mg/mL). At sub-MIC levels, MEW dose-dependently increased doubling times of S. mutans and S. sobrinus up to 258% and 400%, respectively. Furthermore, MEW inhibited acid production, acid tolerance, and biofilm formation of S. mutans and S. sobrinus at sub-MIC levels. The GC-MS analysis revealed the presence of mono- and disaccharides, sugar alcohols, and organic acids as main components. These data suggest that MEW might be useful for restraining physiological activities of cariogenic bacteria.


Assuntos
Antibacterianos/farmacologia , Extratos Vegetais/farmacologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus sobrinus/efeitos dos fármacos , Withania/química , Antibacterianos/química , Antibacterianos/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Cromatografia Gasosa-Espectrometria de Massas , Testes de Sensibilidade Microbiana , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/patogenicidade , Streptococcus mutans/fisiologia , Streptococcus sobrinus/crescimento & desenvolvimento , Streptococcus sobrinus/patogenicidade , Streptococcus sobrinus/fisiologia , Virulência/efeitos dos fármacos
8.
Eur J Paediatr Dent ; 14(2): 125-30, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23758462

RESUMO

AIM: To compare early bacterial adhesion and biofilm formation in vitro by different oral streptococci on a variety of commercial brackets. METHODS: Adhesion and biofilm formation in vitro of 6 Streptococcus spp. on 15 different commercial brackets, in standard culture medium and in human saliva were evaluated by the MTT reduction assay. RESULTS: Significant differences were evidenced in both early adhesion and biofilm formation among the studied brackets and between the two conditions of growth. Gold brackets resulted less prone to colonisation compared to composite brackets. The growth rates of the tested species on the different tested materials were significantly different. CONCLUSION: The adopted experimental plan, dissecting the two phases of plaque formation on different brackets in different conditions, showed that composite brackets are more susceptible to adhesion and colonisation by streptococci, while the remaining tested brackets did not show differences that could be clinically relevant. Data suggest that different personal behaviours affecting the oral environment could significantly affect colonisation of brackets by oral streptococci.


Assuntos
Aderência Bacteriana/fisiologia , Biofilmes/crescimento & desenvolvimento , Boca/microbiologia , Braquetes Ortodônticos/microbiologia , Streptococcus/fisiologia , Óxido de Alumínio/química , Técnicas Bacteriológicas , Cerâmica/química , Corantes , Resinas Compostas/química , Meios de Cultura , Materiais Dentários/química , Placa Dentária/microbiologia , Ligas de Ouro/química , Humanos , Teste de Materiais , Cimento de Policarboxilato/química , Saliva/microbiologia , Aço Inoxidável/química , Streptococcus/classificação , Streptococcus gordonii/fisiologia , Streptococcus mutans/fisiologia , Streptococcus oralis/fisiologia , Streptococcus sanguis/fisiologia , Streptococcus sobrinus/fisiologia , Sais de Tetrazólio , Tiazóis , Titânio/química
9.
Caries Res ; 46(1): 69-77, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22327347

RESUMO

A randomized clinical trial studied the effects of early administration of Bifidobacterium animalis subsp. lactis BB-12 (BB-12) on oral colonization of (1) mutans streptococci (MS), and (2) BB-12. In this double-blind, placebo-controlled study, infants (n = 106) received probiotic bacteria (BB-12 group), xylitol (X group), or sorbitol (S group). Test tablets were administered twice a day (from the age of 1-2 months) with a novel slow-release pacifier or a spoon (daily dose of BB-12 10(10) CFU, polyol 200-600 mg). Samples were collected from mucosa/teeth at the age of 8 months and 2 years for BB- 12 determination (qPCR) and plate culturing of MS (MSB, TYCSB), lactobacilli (Rogosa) and yeasts (Sabouraud). The MS levels of the mothers were determined (Dentocult SM Strip Mutans). The baseline characteristics of the three groups were similar. Mean duration of tablet delivery was 14.9 ± 6.7 months. In all groups, >90% of the mothers showed high MS counts (log CFU ≥5). MS colonization percentages of the children at the age of 2 years were rather low (BB-12 group: 6%; X group: 31%; S group: 10%; p < 0.05). The levels of lactobacilli and yeasts did not differ between the groups. BB-12 cell counts barely exceeding the detection limit were found in three of the oral samples of the 8-month-old children; however, the 2-year samples did not contain BB-12. The early administration of BB-12 did not result in permanent oral colonization of this probiotic or significantly affect MS colonization in the children.


Assuntos
Bifidobacterium/fisiologia , Boca/microbiologia , Probióticos/uso terapêutico , Streptococcus mutans/fisiologia , Carga Bacteriana , Bifidobacterium/isolamento & purificação , Aleitamento Materno , Candida albicans/isolamento & purificação , Candida albicans/fisiologia , Preparações de Ação Retardada , Placa Dentária/microbiologia , Método Duplo-Cego , Feminino , Seguimentos , Humanos , Lactente , Lactobacillus/isolamento & purificação , Lactobacillus/fisiologia , Masculino , Mucosa Bucal/microbiologia , Chupetas , Placebos , Probióticos/administração & dosagem , Sorbitol/administração & dosagem , Sorbitol/uso terapêutico , Streptococcus sobrinus/isolamento & purificação , Streptococcus sobrinus/fisiologia , Comprimidos , Dente/microbiologia , Escovação Dentária , Xilitol/administração & dosagem , Xilitol/uso terapêutico
10.
Am J Orthod Dentofacial Orthop ; 137(4): 489-95; discussion 13A, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20362908

RESUMO

INTRODUCTION: In this study, we analyzed the surface roughness (SR) and surface free energy (SFE) characteristics of various orthodontic adhesives and their effects on the adhesion of mutans streptococci (MS). METHODS: Three nonfluoride-releasing composites, 1 fluoride-releasing composite, 1 polyacid-modified composite (compomer), and 2 resin-modified glass ionomers (RMGIs) were analyzed with confocal laser scanning microscopy and the sessile drop method. Each material was incubated with whole saliva or phosphate-buffered saline solution. Adhesion assays were then performed by incubating each material with tritium-labeled cariogenic streptococci. RESULTS: We found that SFE and its components were significantly different among the adhesives, despite relatively uniform SR. RMGIs showed significantly higher SFE-particularly, dispersive, polar, and acidic characteristics-than did the composites. The compomer had intermediate characteristics between the composites and RMGIs. MS adhered to RMGIs significantly more than to the composites and the compomer, but there was no significant difference among the composites. The longer incubation time increased MS adhesion, whereas saliva coating did not significantly influence MS adhesion. There was a correlation between SFE and MS adhesion. SFE and its dispersive, polar, and acid components significantly influenced MS adhesion irrespective of saliva coating. CONCLUSIONS: This study suggests that initial MS adhesion is significantly influenced by SFE characteristics of adhesives rather than SR or saliva coating.


Assuntos
Aderência Bacteriana/fisiologia , Cimentos Dentários/química , Aparelhos Ortodônticos , Streptococcus/fisiologia , Resinas Acrílicas/química , Silicatos de Alumínio/química , Soluções Tampão , Cariostáticos/química , Compômeros/química , Resinas Compostas/química , Fluoretos/química , Cimentos de Ionômeros de Vidro/química , Humanos , Teste de Materiais , Microscopia Confocal , Fosfatos/química , Cimentos de Resina/química , Saliva/química , Cloreto de Sódio/química , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Propriedades de Superfície , Viscosidade
11.
Acta Odontol Latinoam ; 23(3): 210-5, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21638961

RESUMO

Kinetic hydrophobic measurements were performed by confronting 40 mutans streptococci from thirty 10- to 20-year-old patients with 200 ml hexadecane (Sigma). Fourteen patients had high dental caries risk (Group A), dmft + DMFT > 5 with 3 or more active caries, and 16 had low dental caries risk (Group B), dmft + DMFT < 3 without active caries. Twenty bacteria from Group A and 20 bacteria from Group B were typed using De La Higuera's procedure and confirmed by API strip (bio-Merieux). From the 14 patients in Group A we obtained 12 S. mutans (8 hydrophobic/4 non-hydrophobic), 5 S. sobrinus (4 hydrophobic/1 non-hydrophobic) and 3 S. rattus (hydrophobic). From the 16 patients in Group B we obtained 11 Streptococcus mutans (10 non-hydrophobic/1 hydrophobic), 7 Streptococcus sobrinus (6 non-hydrophobic/ 1 hydrophobic) and 2 Streptococcus rattus (hydrophobic). Patients with high dental caries risk have a higher prevalence of hydrophobic bacteria than patients with low dental caries risk (p = 0.0003). All typed S. rattus were hydrophobic.


Assuntos
Suscetibilidade à Cárie Dentária/fisiologia , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Adolescente , Alcanos/química , Aderência Bacteriana/fisiologia , Criança , Índice CPO , Cárie Dentária/microbiologia , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Streptococcus/classificação , Streptococcus/fisiologia , Adulto Jovem
12.
Oral Microbiol Immunol ; 24(3): 224-30, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19416452

RESUMO

INTRODUCTION: Streptococcus sobrinus exhibits more significant dextran-dependent aggregation mediated by glucan-binding proteins than Streptococcus mutans. We have identified four glucan-binding protein C gene (gbpC) homologues designated as gbpC1, gbpC2, dblA and dblB in S. sobrinus in contrast to the single gene gbpC in S. mutans. We attempted to determine which gene is most responsible for the dextran-dependent aggregation of S. sobrinus. METHODS: We introduced mutation with a chemical mutagen, 1-methyl-3-nitro-1-nitrosoguanidine, into S. sobrinus strain 6715 and analysed the four gbpC homologous gene sequences in the parental strain 6715 and an obtained aggregation-negative mutant NUM-Ssg99. We also examined the localization of proteins encoded by these genes in the mutant NUM-Ssg99. RESULTS: The nucleotide sequences of the gbpC1, gbpC2 and dblA genes in NUM-Ssg99 were 100% identical to the homologous genes in parental strain 6715. In contrast, a truncated mutation was detected in the dblB gene and the mutant protein devoid of the LPXTG motif was confirmed by Western blot analysis to be released into the extracellular milieu. CONCLUSION: We conclude that the dblB gene among the four GbpC homologous protein genes is most responsible for aggregation in strain 6715.


Assuntos
Proteínas de Transporte/genética , Genes Bacterianos/genética , Lectinas/genética , Streptococcus sobrinus/genética , Motivos de Aminoácidos/genética , Aminoaciltransferases/genética , Proteínas de Bactérias/genética , Pareamento de Bases/genética , Sequência de Bases/genética , Western Blotting , Proteínas de Transporte/efeitos dos fármacos , Mapeamento Cromossômico , Cisteína Endopeptidases/genética , Dextranos , Eletroforese em Gel de Poliacrilamida , Humanos , Lectinas/efeitos dos fármacos , Metilnitronitrosoguanidina , Mutagênicos , Mutação/genética , Peptidoglicano/genética , Fenótipo , Ligação Proteica/genética , Análise de Sequência de DNA , Deleção de Sequência/genética , Homologia de Sequência , Streptococcus sobrinus/fisiologia , Transaminases/genética
13.
Eur J Oral Sci ; 117(4): 419-26, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19627354

RESUMO

Bacterial adhesion to biomaterial surfaces constituting the bracket-adhesive-enamel junction represents a growing problem in orthodontics, because bacteria can adversely affect treatment by causing demineralization of the enamel surface around the brackets. It is important to know the forces with which bacteria adhere to the surfaces of these junction materials, as the strength of these forces will determine how easy it will be to remove the bacteria. We compared the adhesion forces of five initially colonizing and four cariogenic strains of bacteria to an orthodontic adhesive, stainless steel, and enamel, with and without a salivary conditioning film. Adhesion forces were determined using atomic force microscopy and a bacterial probe. In the absence of a salivary conditioning film, the strongest bacterial adhesion forces occurred to the adhesive surface (-2.9 to -6.9 nN), while adhesion forces to the enamel surfaces were lowest (-0.8 to -2.7 nN). In the presence of a salivary conditioning film, adhesion forces were reduced strongly, to less than 1 nN, and the differences between the various materials were reduced. Generally, however, initial colonizers of dental hard surfaces presented stronger adhesion forces to the different materials (-4.7 and -0.6 nN in the absence and presence of a salivary conditioning film, respectively) than cariogenic strains (-1.8 and -0.5 nN).


Assuntos
Aderência Bacteriana/fisiologia , Materiais Biocompatíveis/química , Biofilmes , Esmalte Dentário/microbiologia , Boca/microbiologia , Braquetes Ortodônticos/microbiologia , Ortodontia Corretiva/instrumentação , Cimentos de Resina/química , Actinomyces/fisiologia , Animais , Fenômenos Biomecânicos , Bovinos , Ligas Dentárias/química , Película Dentária/microbiologia , Película Dentária/fisiologia , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Lactobacillus acidophilus/fisiologia , Masculino , Teste de Materiais , Aço Inoxidável/química , Streptococcus/fisiologia , Streptococcus mitis/fisiologia , Streptococcus mutans/fisiologia , Streptococcus oralis/fisiologia , Streptococcus sobrinus/fisiologia , Estresse Mecânico , Molhabilidade
14.
Acta Odontol Scand ; 67(5): 284-8, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19479452

RESUMO

OBJECTIVE: To evaluate the in vitro abilities of probiotic bacteria derived from consumer products to coaggregate with caries-associated mutans streptococci. MATERIAL AND METHODS: Six lactobacillus strains (L. acidophilus (CCUG 5917), L. plantarum 299v, L. rhamnosus GG and LB21, L. paracasei F19, L. reuteri PTA5289) were cultivated under anaerobic conditions at 37°C in Man Rogosa Sharpe (MSB) broth for 24 h. Four strains of human streptococci (S. mutans Ingbritt, S. mutans (ATCC 25175), S. mutans GS-5, S. sobrinus (ATCC 33478) were similarly grown in Brain Heart Infusion (BHI) broth. A gastrointestinal pathogen (Escherichia coli) was aerobically cultivated on BHI broth as a positive control. After incubation, the bacteria were aerobically harvested, washed, and suspended in 10 mmol/l phosphate-buffered saline (pH 7.2). The probiotic strains were characterized with the API 50 CH system to confirm their identity. Coaggregation was determined by spectrophotometry in mixtures and bacterial suspensions alone after 1, 2, 4, and 24 h and expressed as the aggregation ratio (%). RESULTS: All probiotic strains showed coaggregation abilities with the oral pathogens and the results were strain specific and dependent on time. S. mutans GS-5 exhibited a significantly higher ability to coaggregate with all the probiotic strains than the other mutans streptococci and E. coli. The differences among the probiotic strains were modest with L. acidophilus being the most prone and L. rhamnosus LB21 the least prone to coaggregate with the oral streptococci. CONCLUSIONS: The results demonstrated different abilities of lactobacilli-derived probiotic bacteria to coaggregate with selected oral streptococci. Aggregation assays may be a useful complement for screening of probiotic candidates with possible anti-caries properties.


Assuntos
Aderência Bacteriana/fisiologia , Lactobacillus/fisiologia , Probióticos , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Anaerobiose , Técnicas Bacteriológicas , Técnicas de Cocultura , Cárie Dentária/microbiologia , Escherichia coli/fisiologia , Humanos , Concentração de Íons de Hidrogênio , Lactobacillus acidophilus/fisiologia , Lactobacillus plantarum/fisiologia , Limosilactobacillus reuteri/fisiologia , Lacticaseibacillus rhamnosus/fisiologia , Espectrofotometria , Fatores de Tempo
15.
Oral Health Prev Dent ; 7(1): 47-53, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19408815

RESUMO

PURPOSE: This laboratory study assessed the influence of surface roughness and contact time on the formation of a multi-species biofilm on dental materials (adhesive patch, composite, amalgam and enamel). MATERIALS AND METHODS: Rough and smooth specimens of each material were prepared and the mean surface roughness was assessed profilometrically. The biofilms were then allowed to grow either for 15 min or 15 h respectively on saliva-preconditioned specimens of each material, and colony-forming units on blood agar were counted (N = 9/group) Surface morphology was assessed using a scanning electron microscope. RESULTS: No difference was found in the biofilm formation rate among all the materials that were tested. After a short incubation period, a statistical significant difference between smooth and rough samples could be detected on amalgam and on the resin composite material (P < or = 0.001). CONCLUSIONS: Surface roughness may influence the initial biofilm adherence, but differences vanish following growth and maturation. The multi-species biofilm offers a reliable laboratory model for studying plaque formation.


Assuntos
Biofilmes/crescimento & desenvolvimento , Materiais Dentários/química , Acrilatos/química , Resinas Acrílicas/química , Actinomyces/fisiologia , Animais , Candida albicans/fisiologia , Materiais Revestidos Biocompatíveis/química , Contagem de Colônia Microbiana , Resinas Compostas/química , Ligas Dentárias/química , Amálgama Dentário/química , Esmalte Dentário/microbiologia , Fusobacterium nucleatum/fisiologia , Teste de Materiais , Microscopia Eletrônica de Varredura , Ácidos Polimetacrílicos/química , Poliuretanos/química , Saliva Artificial/química , Streptococcus oralis/fisiologia , Streptococcus sobrinus/fisiologia , Propriedades de Superfície , Fatores de Tempo , Veillonella/fisiologia
16.
Methods Mol Biol ; 1922: 393-403, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30838593

RESUMO

Dental caries is an infectious oral disease caused primarily by complex interactions of cariogenic oral flora (biofilm) with dietary carbohydrates on the tooth surface over time. Streptococcus mutans and Streptococcus sobrinus (S. mutans and S. sobrinus) are the most prevalent cariogenic species within the oral biofilm and considered the main etiological agents of caries. Pulp exposure and infection can be caused by trauma, carious lesion, and mechanical reasons. Pulp response to these exposures depends on the state of the pulp as well as the potential bacterial contamination of pulp tissue. Herein, we describe the process of using two in vivo rodent models to study the progression of dental caries and pulp disease: a nutritional microbial model and a pulp disease induction model. The progression of the carious lesion and pulpal infections in both models was assessed by micro-CT imaging and histomorphometric analysis. Moreover, the pulp disease induction models can be used to compare and assess the antibacterial and reparative properties of the different pulp capping materials.


Assuntos
Cárie Dentária/microbiologia , Cárie Dentária/patologia , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Animais , Biofilmes , Polpa Dentária/microbiologia , Polpa Dentária/patologia , Modelos Animais de Doenças , Progressão da Doença , Ratos Sprague-Dawley , Microtomografia por Raio-X/métodos
17.
Caries Res ; 42(6): 444-8, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18832831

RESUMO

The aim of this study was to examine the colonization of Streptococcus mutans and Streptococcus sanguinis in the oral cavity and the association with severe early childhood caries (S-ECC). Saliva and plaque samples were collected from 14 S-ECC children and 8 caries-free (CF) children. All S-ECC children were S. mutans positive; 100% of CF children and 93% of S-ECC children were S. sanguinis positive. The children's caries severity was positively correlated with levels of S. mutans (p < 0.001), total oral streptococci (p < 0.01), total cultivable oral bacteria (p < 0.05), and children's age (p < 0.05). Logistic regression analysis showed that the interaction of S. sanguinis with S. mutans was a significant factor associated with the caries status in children, suggesting that the relative levels of these two microorganisms in the oral cavity play an important role in caries development.


Assuntos
Índice CPO , Cárie Dentária/microbiologia , Streptococcus mutans/fisiologia , Streptococcus/fisiologia , Criança , Pré-Escolar , Contagem de Colônia Microbiana , Placa Dentária/microbiologia , Feminino , Humanos , Lactobacillus/fisiologia , Masculino , Saliva/microbiologia , Streptococcus/classificação , Streptococcus sobrinus/fisiologia
18.
Oral Health Prev Dent ; 6(4): 337-41, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-19178100

RESUMO

PURPOSE: The aim of the present study was to examine whether xylitol, at different concentrations, inhibits the formation of an experimental model of oral biofilm. MATERIALS AND METHODS: Biofilms of six bacterial species (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus rhamnosus, Actinomyces viscosus, Porphyromonas gingivalis and Fusobacterium nucleatum) were prepared on hydroxyapatite (HA) discs according to the Zürich Biofilm Model. Xylitol was tested at two concentrations, 1% and 3%. At the end of their designated incubation times, some HA discs were destined for confocal laser scanning microscopy (CLSM) and the others were harvested using a sterile surgical instrument. Aliquots of harvested biofilms were diluted and plated onto specific media. After a 48-h anaerobic incubation at 37 degrees C, the colony-forming units (CFUs) were counted. RESULTS: CLSM images showed that only a small amount of isolated bacteria was observed on the surface of HA discs. Culture of harvested biofilms showed an inhibition in the growth of different species included in the biofilms. CONCLUSIONS: Xylitol has a clear inhibitory effect on the formation of the experimental biofilms. This study shows that xylitol is not only efficient in inhibiting the acid production of cariogenic bacteria, but also in preventing the formation of a multispecies biofilm; it confirms the relevance of the use of this polyol for the prevention of oral diseases caused by dental plaque.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Depósitos Dentários/microbiologia , Bacilos e Cocos Aeróbios Gram-Negativos/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Xilitol/farmacologia , Actinomyces viscosus/efeitos dos fármacos , Actinomyces viscosus/fisiologia , Contagem de Colônia Microbiana , Relação Dose-Resposta a Droga , Durapatita , Fusobacterium nucleatum/efeitos dos fármacos , Fusobacterium nucleatum/fisiologia , Bacilos e Cocos Aeróbios Gram-Negativos/fisiologia , Bactérias Gram-Positivas/fisiologia , Lacticaseibacillus rhamnosus/efeitos dos fármacos , Lacticaseibacillus rhamnosus/fisiologia , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/fisiologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologia , Streptococcus sobrinus/efeitos dos fármacos , Streptococcus sobrinus/fisiologia , Dente Artificial/microbiologia
19.
Am J Orthod Dentofacial Orthop ; 133(6): 882-8, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18538253

RESUMO

INTRODUCTION: Knowledge of adhesion patterns of cariogenic streptococci to orthodontic materials can provide valuable information on the cause of enamel demineralization during orthodontic treatment. The purpose of this study was to investigate the adhesion of 2 cariogenic streptococci strains to 7 orthodontic raw materials (3 light-cured orthodontic adhesives, 3 bracket raw materials, and hydroxyapatite) with respect to bacterial species, incubation time, and saliva coating. METHODS: Each material was incubated with unstimulated whole saliva or phosphate-buffered saline solution for 2 hours. Binding assays were then performed by incubating tritium-labeled cariogenic streptococci with each raw material for 3 or 6 hours. RESULTS: The degree of adhesion varied by material type. Generally, adhesion of cariogenic streptococci was significantly higher for bonding adhesives than for bracket materials, and adhesion to resin-modified glass ionomer was the highest. A longer incubation time generally increased bacterial adhesion, whereas saliva coating did not significantly influence bacterial adhesion. CONCLUSIONS: Bonding adhesives around brackets should be removed carefully during the bonding procedure to avoid enamel decalcification.


Assuntos
Aderência Bacteriana , Aparelhos Ortodônticos/microbiologia , Streptococcus mutans/fisiologia , Streptococcus sobrinus/fisiologia , Adulto , Óxido de Alumínio , Análise de Variância , Cerâmica , Durapatita , Cimentos de Ionômeros de Vidro , Humanos , Cimentos de Resina , Saliva/microbiologia , Aço Inoxidável
20.
Sci Rep ; 8(1): 3767, 2018 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-29491366

RESUMO

Sealing can arrest caries lesions. We aimed to evaluate if sealing effects and kinetics are bacterial-strain and sealing-material specific. Human dentin discs were mounted in a dual-chamber device. Caries lesions were induced chemically and contaminated with either Lactobacillus rhamnosus (LR) or Streptococcus sobrinus (SS). For (1) kinetics assessment, the initial bacterial load and the sealing period were varied, and lesions sealed using a self-etch adhesive and composite. For (2) comparing materials, six sealing protocols (#1-#6) were evaluated: 1# Self-etch adhesive plus composite placed without a liner, or #2 calcium hydroxide, or #3 mineral trioxide aggregate, or #4 Biodentine liners; #5 antibacterial adhesive plus composite; #6 glass ionomer cement. Pulpal fluid flow was simulated during sealing. The outcome was the number of surviving bacteria (CFU) per g dentin. For LR, bacterial survival increased significantly with increasing initial bacterial load and decreased with longer sealing periods. The relative reduction followed a first-order kinetics. More LR survived under calcium hydroxide or MTA than other materials (p < 0.001). For SS, nearly no bacteria survived sealing regardless of sealing period, initial bacterial load or sealing material. In conclusion, sealing effects and kinetics were strain- and material-specific.


Assuntos
Cárie Dentária/microbiologia , Lacticaseibacillus rhamnosus/efeitos dos fármacos , Cimentos de Resina/farmacologia , Streptococcus sobrinus/efeitos dos fármacos , Humanos , Cinética , Lacticaseibacillus rhamnosus/fisiologia , Especificidade da Espécie , Streptococcus sobrinus/fisiologia
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