Efectos del xilitol en el crecimiento bacteriano frente a Streptococcus sanguinis: Estudio in vitro

Streptococcus sanguinis forma parte del biofilm bucal, tiene función decisoria en el desarrollo de las enfermedades bucales prevalentes y a nivel sistémico actúa como patógeno oportunista. Objetivo: Evaluar in vitro los efectos del xilitol en el crecimiento bacteriano frente a Streptococcus sanguinis (ATCC 10556). Métodos: la muestra del estudio fue distribuida en 6 grupos: 4 grupos experimentales (xilitol 1M; 0,75M; 0,50M y 0,25M), un control negativo (agua destilada) y un control positivo (clorhexidina); el análisis estadístico se hizo mediante el software estadístico Infostat y se empleó las pruebas t-Student, ANOVA y Tukey para contrastar la hipótesis. Resultados: diferentes concentraciones de xilitol (0,25M; 0,50M; 0,75M y 1M) causaron un halo de inhibición entre 9,89 - 12,89 mm (24 horas) y 10,85 - 13,45 mm (48 horas). Conclusiones: diferentes concentraciones de xilitol inhiben el crecimiento bacteriano del Streptococcus sanguinis, este efecto inhibitorio aumenta a mayor concentración y tiempo de exposición.

Streptococcus sanguinis faz parte do biofilme oral, tem papel decisivo no desenvolvimento de doenças bucais prevalentes e atua como patógeno oportunista em nível sistêmico. Objetivo: Avaliar in vitro os efeitos do xilitol no crescimento bacteriano contra Streptococcus sanguinis (ATCC 10556). Métodos: A amostra do estudo foi distribuída em 6 grupos: 4 grupos experimentais (1M; 0,75M; 0,50M e 0,25M xilitol), um controle negativo (água destilada) e um controle positivo (clorexidina); a análise estatística foi feita com o software estatístico Infostat e os testes t-Student, ANOVA e Tukey para testar a hipótese. Resultados: diferentes concentrações de xilitol (0,25M; 0,50M; 075M e 1M) causou um halo de inibição entre 9,89 - 12,89 mm (24 horas) e 10,85 - 13,45 mm (48 horas). Conclusões: diferentes concentrações de xilitol inibem o crescimento bacteriano de Streptococcus sanguinis, este efeito inibitório aumenta com maior concentração e tempo de exposição.

Streptococcus sanguinis forms part of the oral biofilm, has a decisive role in the development of prevalent oral diseases and acts as an opportunistic pathogen at the systemic level. Aims: To evaluate in vitro the effects of xylitol on bacterial growth against Streptococcus sanguinis (ATCC 10556). Methods: The study sample was distributed into 6 groups: 4 experimental groups (1M; 0,75M; 0,50M and 0,25M xylitol), a negative control (distilled water) and a positive control (chlorhexidine). The statistical analysis was done using the statistical software Infostat and the tests used t-Student, ANOVA and Tukey to test the hypothesis. Results: different concentrations of xylitol (0,25M; 0,50M; 0,75M and 1M) caused an inhibition halo between 9,89 - 12,89 mm (24 hours) and 10,85 - 13,45 mm (48 hours). Conclusions: different concentrations of xylitol inhibit the bacterial growth of Streptococcus sanguinis, this inhibitory effect increases with higher concentration and exposure time.

Aplicação do plasma de baixa temperatura sob pressão atmosférica no controle de biofilmes cariogênicos / Application of low temperature plasma under atmospheric pressure in the control cariogenic biofilms

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU) The general objective of this study was to evaluate the application of lowtemperature plasma under atmospheric pressure (PBTPA) of argon and helium flow, in the control of cariogenic biofilms. For this, the effective physical parameters of PBTPA-argon and helium in mono, dual and polymicrobial biofilms composed of combinations of the species Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans and Actinomyces naeslundii were established. The multi-species biofilms were treated by different PBTPA generating devices, one obtained commercially (kINPen09®) that used argon as working gas, and another prototype developed by FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) that used helium as working gas. Quantitative and microscopic analyzes (confocal, scanning electron microscopy) were performed. Negative control (no treatment), positive control (chlorhexidine 2%) and gas control (argon) were included. Besides that, cellular effects of PBTPA-argon and helium on dual and multi-species biofilms were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy. The results obtained for the treatments of mono, dual or multispecies biofilms with both PBTPA-argon and helium were all significant when compared to the negative control at all times analyzed. For PBTPA-argon, there was no recovery of S. gordonii and S. sanguinis at all analyzed times. For PBTPA-helium, the best results were obtained at 5 and 7 min of exposure of biofilms to PBTPA. All the tests were carried out in triplicate in three independent experiments. The results are tabulated and analyzed in terms of distribution. Next, the most suitable statistical tests were selected. The level of significance was 5%. The results obtained for the treatments of mono, dual or multi-species biofilms with PBTPA-argon and helium were all significant compared to the negative control at all analyzed times. Finally, both PBTPA generating could contribute to the development of new protocols for Minimal Intervention Dentistry (AU)

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU)

Inhibition Effect of Garlic (Allium sativum) Extract on Streptococcus sanguinis Biofilm Formation Involving Bacterial Motility Mechanism

@#Introduction: Streptococcus sanguinis is a primary colonizer in oral biofilm formation, often implicated in infective endocarditis. Methods to control oral biofilm formation are yet to be developed. Garlic (Allium sativum) has shown antimicrobial activities against many pathogen species. We sought to observe the potential of garlic extract to inhibit bacterial adherence to hydroxyapatite (HA) discs as a model of the tooth surface. Methods: Susceptibility of S. sanguinis ATCC 10556 to garlic extract was examined by minimum inhibitory concentration (MIC) test using broth microdilution method. For bacterial adherence assay, saliva-coated HA discs were incubated with various concentrations of extract then stimulated with S. sanguinis ATCC 10556 suspension. Adherent bacteria were stained with 0.1% crystal violet and measured at 595 nm using a microplate reader. A qualitative method to test bacterial motility was performed using Motility Indole Ornithine (MIO) medium. Results: The result of minimum inhibitory concentration test showed that MIC value for garlic ethanolic extract was at a concentration of 625 μg/ml. Moreover, garlic extract inhibited bacterial adherence to HA discs starting at concentration of 62.5 μg/ml. The inhibition of bacterial motility can be observed, indicated as limited the diffused growth of bacteria closer to the inoculating line. Observation using SEM confirmed these results. Conclusion: This present study suggest that garlic extract has the ability to inhibit S. sanguinis adherence to HA discs involving inhibition of bacterial motility, with the optimal concentration being 500 µg/ml.

Effects of antimicrobial peptide GH12 on the morphology and composition of cariogenic three-species biofilm / 华西口腔医学杂志

OBJECTIVES@#To evaluate the effects of antimicrobial peptide GH12 designed @*METHODS@#The cariogenic three-species biofilm consis-ted of the cariogenic @*RESULTS@#The biomass and density of the cariogenic three-species biofilm treated with GH12 decreased compared with those of the control. The number of @*CONCLUSIONS@#GH12 can reduce the number of

Influence of environmental factors on the two-species biofilm formed by Streptococcus oligofermentans and Streptococcus mutans / 中华口腔医学杂志

Objective@#To study the influence of environmental factors on the two-species biofilm formed by the combinations of Streptococcus oligofermentans (So) with Streptococcus mutans (Sm) and Streptococcus sanguinis (Ss) with Sm so as to evaluate the role of So in maintaining the microecological balance of the oral cavity.@*Methods@#Single-and two-species biofilms were grown on saliva-coated surfaces (glass tube and 96-well plate). Colony-counting method and safranin staining method were used to measure the biofilms formed under various oxygen conditions (aerobic and anaerobic), sucrose conditions (0%, 1% and 5% sucrose concentrations) and pH conditions (5.5, 6.0, 6.5, 7.0, 7.5 and 8.0).@*Results@#Comparing the numbers of Sm in two co-cultures under various conditions, Sm counts in So+Sm group [(7.70±2.46)×108 CFU/ml] were significantly lower than those in Ss+Sm group [(9.00±1.13)×108 CFU/ml] in aerobic environment (P<0.05). Sm counts in So+Sm group [(2.80±0.52)×108 CFU/ml] were also significantly lower than those in the Ss+Sm group [(4.00±1.25)×108 CFU/ml] in anaerobic environment (P<0.05). The Sm counts in So+Sm group [(8.90±0.82)×108 CFU/ml] were significantly higher than those in Ss+Sm group [(7.50±1.73)×108 CFU/ml] in 0% sucrose environment (P<0.05). The Sm counts in So+Sm group [(5.70±2.94)×108 CFU/ml] were significantly lower than those in Ss+Sm group [(10.30±3.21) ×108 CFU/ml] in 1% sucrose environment (P<0.05). The Sm counts in So+Sm group [(6.10±1.71)×108 CFU/ml] were also significantly lower than those in Ss+Sm group [(7.40±1.20)×108 CFU/ml] in 5% sucrose environment (P<0.05). The Sm counts in So+Sm group [(3.50±1.50)×108 CFU/ml] were significantly lower than those in Ss+Sm group [(10.70±2.80)×108 CFU/ml] in pH7.0 environment (P<0.05). Comparing the formation of biofilm after 24 h cultivation, the Sm counts in So+Sm group were significantly lower than those in Ss+Sm group both in aerobic and anaerobic environments (P<0.05). The Sm counts in So+Sm group were significantly higher than those in Ss+Sm group in 0% sucrose environment (P<0.05). The Sm counts in So+Sm group were significantly lower than those in Ss+Sm group in 1% and 5% sucrose and pH 7.0 environments (P<0.05). Both So and Ss had no inhibitory effect on Sm in pH5.5 and pH8.0 environments.@*Conclusions@#In the in vitro two-species co-culture systems, So showed stronger inhibitory effects than Ss on Sm and its inhibitory ability might influenced by various environmental factors.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm

Abstract Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. Objective The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual-species biofilm and to evaluate its potential ability on affecting dental health. Material and methods The effect of H. pylori supernatant on single-species and dual-species biofilm was measured by colony forming units counting and fluorescence in situ hybridization (FISH) assay, respectively. The effect of H. pylori supernatant on S. mutans and S. sanguinis extracellular polysaccharides (EPS) production was measured by both confocal laser scanning microscopy observation and anthrone-sulfuric acid method. The effect of H. pylori supernatant on S. mutans gene expression was measured by quantitative real-time PCR (qRT-PCR) assays. Results H. pylori supernatant could inhibit both S. mutans and S. sanguinis biofilm formation and EPS production. S. sanguinis inhibition rate was significantly higher than that of S. mutans. Finally, S. mutans bacteriocin and acidogenicity related genes expression were affected by H. pylori culture supernatant. Conclusion Our results showed that H. pylori could destroy the balance between S. mutans and S. sanguinis in oral biofilm, creating an advantageous environment for S. mutans, which became the dominant bacteria, promoting the formation and development of dental caries.

Streptococcus sanguinis Endocarditis Involving All Valves in a Patient with Ventricular Septal Defect

No abstract available.

Uso odontológico de propóleos de origen costarricense

El objetivo de la investigación fue determinar el nivel de efectividad antimicrobiana in vitro de propóleos altos en compuestos fenólicos de origen costarricense sobre las especies streptococcus sanguinis y streptococcus mutans. Se midió la acción bacteriostática o bactericida del propóleo en una concentración al 50%, 70% y 80%, comparando su efecto contra el digluconato de clorhexidina al 0,12%. El análisis evidenció que el propóleo ejerce una acción bactericida sobre la especie streptococcus sanguinis independientemente de su concentración, y por otra parte sobre la bacteria streptococcus mutans las concentraciones del propóleo al 50% y 70% resultaron en acción bacteriostática. De forma tal se concluye que los tres extractos del propóleo generaron un efecto antimicrobiano sobre las especies S. mutans y S. sanguinis. Se obtuvieron efectos bactericidas de los extractos del propóleo similares al gluconato de clorhexidina al 0,12%, y esto justifica que puede ser empleado como herramienta para la prevención o coadyuvante del tratamiento de la enfermedad periodontal y reducción del riesgo de caries.

The aim of the research was to determine the In Vitro level of antimicrobial effectiveness of Costa Rican propolis high in phenolic compounds on the species Streptococcus Mutans and Streptococcus Sanguinis. It was measured the bacteriostatic and / or bactericidal action of propolis in 50%, 70% and 80% concentration, comparing their effect against chlorhexidine 0.12%. The analysis showed that propolis proved a bactericidal effect on the species Streptococcus Sanguinis regardless of their concentration; on the other hand on the propolis concentrations of 50% and 70% on the bacterium Streptococcus mutans resulted in bacteriostatic action. It is concluded that the three extracts of propolis generated an antimicrobial effect on the species S. mutans and S. Sanguinis. Antibacterial effects from extracts of propolis were similar to chlorhexidine 0.12%, this justifies that can be used as a tool for prevention and / or adjunctive treatment of periodontal disease and reduction of tooth decay risk.

Bactericidal effect of a novel peptide on oral microorganisms / 医学研究生学报

Objective Antimicrobial peptides are the focus of recent research in oral microbiology .This study aimed to eval-uate the activity of a novel antimicrobial peptide pm 11 against oral microorganisms and its action mechanisms . Methods We ana-lyzed the effect of pm11 on oral microorganisms and determined its antimicrobial activity in the saliva environment by measuring its min -imal inhibitory concentration (MIC), minimal bactericide concentration (MBC), and bactericidal kinetics.We observed its bacteri-cidal activity on the biofilms of streptococcus mutans by confocal laser scanning microscopy (CLSM) and the structural changes in the bacterial membrane by scanning electron microscopy (SEM). Results The antimicrobial activity of pm11 varied greatly against dif-ferent oral microorganisms , with its MIC values ranging from 2 μg/mL to 256 μg/mL and its MBC values from 2 μg/mL to >256μg/mL.The bactericidal kinetics showed a decreasing survival rate of bacteria with the lengthening of the intervention time .The inhib-itory-zone diameters exhibited no significant indifference between the water solution and the sterile saliva solution .CLSM revealed an increased number of dead bacteria in the pm 11-treated biofilms , while SEM manifested obvious changes in the shape of the bacteria membrane treated with pm11. Conclusion Our findings suggest that pm11 has a broad spectrum of antimicrobial activities on oral mi-croorganisms and a potential value of clinical application .

Influence of time, toothpaste and saliva in the retention of Streptococcus mutans and Streptococcus sanguinis on different toothbrushes

Objectives: The intraoral transmission of cariogenic and periodontopathogenic species seems to be facilitated by contaminated toothbrushes and other oral hygiene devices. The aim of this investigation was to analyze the in vitro retention and survival rate of Streptococcus mutans and Streptococcus sanguinis on different toothbrushes. The impacts of human saliva and antimicrobial toothpaste on these parameters were further evaluated. Material and Methods: Part I: Four toothbrushes (Colgate 360°, Curaprox CS5460 ultra soft, elmex InterX, Trisa Flexible Head3) were contaminated by S. mutans DSM 20523 or S. sanguinis DSM 20068 suspensions for three minutes. Bacteria were removed from the toothbrushes after either three minutes (T0) or 24 hours (T24) of dry storage and grown on Columbia blood agar plates for the quantification of colony-forming units (CFUs). Part II: The effects of saliva from a caries-active or a caries-inactive person and of toothpaste containing 0.12% chlorhexidine digluconate were also tested. Results: Part I: After three minutes of dry storage, approximately one percent of the bacteria were still detectable on the toothbrushes. After 24 hours, S. sanguinis exhibited a more pronounced decrease in viable cell numbers compared with S. mutans but the differences were not significant (Kruskal-Wallis test, p>0.05). Part II: The addition of human saliva from a caries-active or caries-inactive person slightly increased the retention of both streptococcal species at T0. The use of toothpaste had no influence on the amount of viable streptococci at T0, but it reduced the microbial load after 24 hours of storage. There were only slight nonsignificant differences (p>0.05) between the four toothbrushes. Conclusions: In vitro bacterial retention and survival of S. sanguinis and S. mutans on different toothbrushes occurred. Within the limitations of this study, the use of human saliva or an antimicrobial toothpaste ...

Efecto del ácido lipoteicoico sobre la expresión de genes en cardiomiocitos de ratón (H9c2) / Effect of lipoteichoic acid on gene expression in mice (H9c2) cardiomyocite

La mayoría de las enfermedades de la pulpa dental y de los tejidos perirradiculares guardan relación con microorganismos. Los peptidoglucanos y el ácido lipoteicoico son dos de los principales componentes de las bacterias Gram positivas que tienen actividades relacionadas con el desarrollo de sepsis. Tras la invasión microbiana de estos tejidos, el huésped responde con defensas tanto inflamatorias inespecíficas como inmunológicas específicas. El tratamiento endodóncico quirúrgico y no quirúrgico son en esencia, procedimientos de desbridamiento destinados a destruir y eliminar el ecosistema microbiano, asociado con el proceso patológico. Es importante que los clínicos comprendan la íntima relación entre presencia de microorganismos y enfermedad endodóncica, con el fin de diseñar un tratamiento racional y efectivo; sobre todo en aquellos individuos susceptibles a Endocarditis Infecciosa. En el presente estudio se investigó la expresión de TNFα, IL-1 y COX-2 por efecto del ácido lipoteicoico (ALT) de Streptococcus sanguinis caracterizando las señales intracelulares involucradas en cardiomiocitos H9c2. La línea celular fue tratada con ALT a diferentes concentraciones durante 30 minutos. Comparadas con los controles, las respuestas al tratamiento con ALT fueron dependientes de la dosis y mediante un análisis de One Step RT-PCR (Invitrogen) se evaluó dicha expresión; la cual se asemeja a la respuesta fisiológica del organismo durante un episodio de Endocarditis infecciosa y a la agudización durante un procedimiento endodóncico.

Most dental pulp diseases and diseases of tissues surrounding the root are somehow related to micro-organisms. Peptidoglycans and lipoteichoic acid are two of the main Gram-positive bacteria components with activities related to sepsis development. When tissues sustain microbial invasion the host responds with both unspecific inflammatory defenses and specific immunological reactions. Surgical and non surgical endodontic treatments are essentially debridement procedures intended to destroy and eliminate the microbial eco-system associated to the pathological process. It is essential for clinicians to understand the intimate relationship existing between micro-organisms and endodontic disease, so as to be able to tailor a rational and effective treatment especially in subjects susceptible to infective endocarditis processes. In the present study research was conducted on TNFα, IL-1 COX-2 expression through the effect of lipoteichoic acid (LTA) of Streptococcus sanguinis by characterizing intra-cellular signals involved in H9c'' cardiomyocytes. The cell line was treated with LTA at different concentrations during 30 minutes. When compared to control group, responses to LTA treatment were dependent on dosage. That expression was assessed by means of a One Step RT-PCR (Invitrogen) analysis. It was noted that the aforementioned expression resembled the organisms's physiological response during an infective endocarditis episode and to exacerbation observed during an endodontic procedure.

Development of Streptococcus sanguinis-, Streptococcus parasanguinis-, and Streptococcus gordonii-PCR Primers Based on the Nucleotide Sequences of Species-specific DNA Probes Screened by Inverted Dot Blot Hybridization

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

Formação de biofilme in vitro por Streptococcus sanguinis e Candida albicans, associados, em implantes dentários submetidos a diferentes tratamentos de superfície / In vitro adherence of Streptococcus sanguinis and Candida albicans to dental implants with different surface treatment

O objetivo deste estudo foi avaliar a formação de biofilme in vitro por Streptococcus sanguinis e Candida albicans, associados, em implantes dentários com diferentes tratamentos de superfícies. Foram utilizadas cepas padrão de Streptococcus sanguinis (ATCC10556) e Candida albicans (ATCC 18804). Dos 30 implantes utilizados, dez possuíam superfície lisa (SL), dez foram tratados com duplo ataque ácido (AA) e dez com nanopartículas de hidroxiapatita (NH). Os implantes foram imersos em saliva humana, centrifugada e filtrada, por 60 minutos. Posteriormente, foram transferidos para 1,5 ml de caldo sacarosado e inoculados com 0,1 ml de suspensão de Streptococcus sanguinis (106cél/mL) e incubados em estufa com 5% de CO2 a 37°C. Após 24 horas, os implantes foram transferidos para novo caldo, inoculados com suspensão de Candida albicans (106cél/mL) e incubados por mais 24 horas a 5% de CO2 a 37°C. Os implantes foram lavados e os microrganismos desprendidos em solução fisiológica em agitador ultrassônico. Foram realizadas diluições e alíquotas semeadas em placas com ágar Sabouraud com cloranfenicol e ágar Mitis Salivarius acrescido de bacitracina (0,2 UI/mL) e sacarose (15%), para o crescimento, respectivamente, das leveduras e bactérias, e incubadas a 37°C/48 h. Os números de UFC/ml em log10 foram analisados estatisticamente (Anova, teste de Tukey, p < 0.05). A bactéria Streptococcus sanguinis apresentou maior índice de aderência, porém, sem diferença estatisticamente significante entre os implantes. A aderência de Candida albicans, foi menor nos implantes tratados por NH, com diferença estatisticamente significante em relação os implantes de SL (p = 0,012) e de AA (p = 0,000).

The purpose of this study was to evaluate in vitro adherence of Streptococcus sanguinis and Candida albicans to dental implants with different surface treatment. Standard strains of Streptococcus sanguinis (ATCC10556) and Candida albicans (ATCC 18804) were used. Of the 30 implants used, 10 had a smooth surface (SS), 10 were treated with double acid attack (AA), and 10 with hydroxyapatite nanoparticles (NH). The implants were immersed in human saliva, centrifuged and filtered for 60 minutes. Subsequently, were transferred to 1.5 mL of Gibbons and Nygaard broth and inoculated with 0.1 ml of Streptococcus sanguinis (106cells/mL), and incubated in an incubator with 5% CO2 at 37°C. After 24 h, the implants were transferred to new broth, inoculated with the suspension of Candida albicans (106cells/mL), and incubated for another 24h with 5% CO2 at 37°C. The implants were washed in sterile saline solution in order to remove loosely bound material. The implants were placed into tubes with sterile saline solution and sonicated for 30s. Ten-fold serial dilutions were carried out and aliquots plated on Sabouraud agar with chloramphenicol and Mitis salivarius agar with bacitracin (0.2 IU/mL) and sucrose (15%) for growth, respectively, of yeast and bacteria, and incubated at 37°C/48 h. Then, the numbers CFU/mL (log10) were counted and analyzed statistically (Anova, Tukey´s test, p < 0.05). It was concluded that Streptococcus sanguinis had a higher rate of adhered cells, but with no statistically significant difference among implants. The adherence of Candida albicans was lower in the implants treated with NH, being statistically significant compared to SL (p = 0.012) and AA (p = 0.000) implants.

Aderência in vitro de streptococcus sanguinis e candida albicans em implantes dentários de superfície lisa ou tratada / Adherence in vitro of Streptococcus sanguinis and Candida albicans in dental implants of surface smooth or treated

O objetivo desse trabalho foi analisar in vitro a aderência de Streptococcus sanguinis, Candida albicans e associações destes microrganismos com Streptococcus mutans às superfícies dos implantes dentários tratados com jateamento de fosfato de cálcio, anodização, duplo ataque ácido e os de superfície lisa, com ou sem a prévia incubação em saliva ou plasma sanguíneo. Foram selecionados 120 implantes, sendo 30 de cada superfície para cada microrganismo e associações estudadas. Para análise da aderência, foram preparadas suspensões de microrganismos contendo 106 células/ml em espectrofotômetro. Além disso, cada microrganismo e associações foram divididos em três grupos: em um, o implante foi removido da embalagem e colocado diretamente no caldo, em outro, foi previamente embebido em saliva humana por uma hora e no último em plasma humano, também por uma hora. Os implantes foram acondicionados separadamente em poços de placas de cultura de células contendo caldo sacarosado (placa in vitro) e a suspensão do microrganismo. Após 24h de incubação a 37°C e 5% de CO2, os implantes foram lavados três vezes durante um minuto em solução salina estéril e colocados em sonicador com 10 ml de salina para dispersão das células aderidas. A seguir, foram realizadas diluições seriadas e semeaduras em meios de cultura específico para cada microrganismo. Após 48h de incubação a 37°C e 5% de CO2, foi realizada a contagem de unidades formadoras de colônias (UFC/ml) e os dados foram submetidos a análise de variância (ANOVA), teste de Tukey, com nível de significância de 5%. Para ilustrar a aderência dos microrganismos, foram selecionados o microrganismo Streptococcus sanguinis, e as associações Streptococcus sanguinis e Candida albicans e Streptococcus sanguinis, Streptococcus mutans e Candida albicans foram submetidos à microscopia eletrônica de varredura...

The aim of this study has been to analyze, in vitro the adherence of Streptococcus sanguinis, Candida albicans and associations of those microorganisms with Streptococcus mutans to the surfaces of dental implants treated with calcium phosphate jetting, anodization, double acid attack and to those of smooth surface, with or without previous saliva or blood plasma incubation. Ten implants from each surface were selected for every studied microorganism and association. In order to analyze the adherence, suspensions of microorganisms bearing 106 viable cells/ml in spectrophotometer were prepared. Additionally, every microorganism and association was divided in three groups: in the first, an implant was removed from its wrap and put right away into the sauce; the second, it was previously drenched into saliva for one hour; and the last one, into plasma, for one hour, as well. The implants were separately placed in culturing plaque wells of cells containing saccharose sauce (in vitro plaque) and the microorganism’s suspension. After 24 hours of incubation time at 37 °C and 5% of CO2, the implants were taken washed three times for a minute in saline sterile solution and put in a sonicator holding 10 ml of saline in order to disperse adherent cells. Then, seriated dilutions were done, and sowing in culture media specific for each of the. After a 48h-incubation time at 37°C and 5% of CO2, a counting was carried, of the colony forming units (UFC/ml) and the data were submitted to the ANOVA, Tukey test, at a significance level of 5%. To illustrate the adherence of the microorganisms, some samples were exposed to electronic sweeping microscopy. The results did show great microorganism adherence to the surfaces studied, mainly when associated forming a biofilm...