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.

Effect of roselle calyx extract on in vitro viability and biofilm formation ability of oral pathogenic bacteria

OBJECTIVE@#To investigate the effect of the roselle calyx extract (RCE) (Hibiscus sabdariffa L.) on the in vitro viability and biofilm formation ability of oral pathogenic bacteria.@*METHODS@#RCE was prepared by soaking roselle calyx powder with ethyl alcohol for 24 h at room temperature. After centrifugation, the extract was lyophilized. Then, the extract was dissolved in phosphate-buffered saline, the pH was adjusted, and the extract was aseptically filtered. We used Streptococcus mutans, Streptococcus sanguinis, Lactobacillus casei, Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in this study. The antibacterial activity of the RCE was determined by treating the cells of these bacteria with the extract for 10 or 20 min at room temperature. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration was determined using the microdilution method, and the effect of the RCE on the ability to form biofilm was determined using a polystyrene micro plate assay. In addition, we used the WST-1 assay to determine the cytotoxicity of the RCE on HGF, Ca9-22 and KB cells.@*RESULTS@#The RCE had antibacterial activity against oral bacteria used in this study. In particular, most significant antibacterial activity was observed against Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas gingivalis. The MIC and minimum bactericidal concentration were 7.2 mg/mL-28.8 mg/mL and 14.4 to >57.6 mg/mL. The RCE had an inhibitory effect on biofilm formation at the MIC and sub-MIC levels. In addition, the RCE had low cytotoxic effects on HGF, Ca9-22 and KB cells.@*CONCLUSIONS@#Thus, our results indicate that the RCE may be used for preventing oral diseases.

Effect of roselle calyx extract on in vitro viability and biofilm formation ability of oral pathogenic bacteria

Objective: To investigate the effect of the roselle calyx extract (RCE) (Hibiscus sabdariffa L.) on the in vitro viability and biofilm formation ability of oral pathogenic bacteria. Methods: RCE was prepared by soaking roselle calyx powder with ethyl alcohol for 24 h at room temperature. After centrifugation, the extract was lyophilized. Then, the extract was dissolved in phosphate-buffered saline, the pH was adjusted, and the extract was aseptically filtered. We used Streptococcus mutans, Streptococcus sanguinis, Lactobacillus casei, Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in this study. The antibacterial activity of the RCE was determined by treating the cells of these bacteria with the extract for 10 or 20 min at room temperature. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration was determined using the microdilution method, and the effect of the RCE on the ability to form biofilm was determined using a polystyrene micro plate assay. In addition, we used the WST-1 assay to determine the cytotoxicity of the RCE on HGF, Ca9-22 and KB cells. Results: The RCE had antibacterial activity against oral bacteria used in this study. In particular, most significant antibacterial activity was observed against Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas gingivalis. The MIC and minimum bactericidal concentration were 7.2 mg/mL-28.8 mg/mL and 14.4 to >57.6 mg/mL. The RCE had an inhibitory effect on biofilm formation at the MIC and sub-MIC levels. In addition, the RCE had low cytotoxic effects on HGF, Ca9-22 and KB cells. Conclusions: Thus, our results indicate that the RCE may be used for preventing oral diseases.

Comparison of Antimicrobial Activity of Electrolyzed Water Using Various Electrodes against Biofilm of Oral Pathogens

Biofilms of oral microbes can cause various diseases in the oral cavity, such as dental caries, periodontitis and mucosal disease. Electrolyzed water generated by an electric current passed via water using a metal electrode has an antimicrobial effect on pathogenic bacteria which cause food poisoning. This study investigated the antimicrobial activity of electrolyzed waters using various metal electrodes on the floatage and biofilms of oral microbes. The electrolyzed water was generated by passing electric current using copper, silver and platinum electrodes. The electrolyzed water has a neutral pH. Streptococcus mutans, Porphyromonas gingivalis and Tannerella forsythia were cultured, and were used to form a biofilm using specific media. The floatage and biofilm of the microbes were then treated with the electrolyzed water. The electrolyzed water using platinum electrode (EWP) exhibited strong antimicrobial activity against the floatage and biofilm of the oral microbes. However, the electrolyzed water using copper and silver electrodes had no effect. The EWP disrupted the biofilm of oral microbes, except the S. mutans biofilm. Comparing the different electrolyzed waters that we created the platinum electrode generated water may be an ideal candidate for prevention of dental caries and periodontitis.

Cuantificación de bacterias relacionadas con la caries dental en saliva de adultos y adultos mayores / Quantification of caries-associated bacteria from saliva of adults and older adults

Introducción: Se considera que Streptococcus mutans (S. mutans) y Lactobacillus spp. se asocian con la caries. Otras especies del biofilm oral, como Streptococcus sanguinis (S. sanguinis) han sido sindicadas como protectoras, pero principalmente en niños. Existe escasa evidencia sobre el nivel de estas bacterias en adultos mayores. Objetivo: Determinar si existen diferencias en los recuentos microbianos de tres especies relacionadas con la caries en pacientes adultos y adultos mayores. Materiales y Métodos: Una muestra de pacientes por conveniencia compuesta de 63 pacientes (18 a 79 años) proporcionó saliva estimulada con la que se sembraron placas de agar MSB, MM10 SB y Agar Rogosa para el cultivo de S. mutans, S. sanguinis y Lactobacillus spp., respectivamente. Los recuentos bacterianos fueron expresados como UFC/mL. Resultados: Los recuentos de S. mutans y Lactobacillus spp. no mostraron variaciones relacionadas con la edad (p>0.05). Los adultos mostraron mayores recuentos de S. sanguinis que los adultos mayores, 3.7 x 105 +/- 3.8 x 105 UFC/mL y 5.9 x 104 +/- 9.4 x 104 UFC/mL, respectivamente (p<0.05). Conclusiones: La edad no parece afectar los niveles de especies tradicionalmente consideradas como cariogénicas. Estos resultados sugieren que la edad puede relacionarse con los patrones de colonización de S. sanguinis en el biofilm oral.

Introduction: Streptococcus mutans (S. mutans) and Lactobacillus spp. have been traditionally associated with caries, regardless of the subject’s age. Other oral biofilm species have been linked as caries protective, including Streptococcus sanguinis (S. sanguinis), but mainly in children. Scarce evidence exists on the levels of these bacteria in older adults. Aim: To determine whether there are differences in the microbial counts of three caries-associated bacterial species in adults and older adults. Methodology: A convenience sample of sixty three patients, aged 18 to 79 years, participated in the study. Stimulated saliva samples were obtained and in MSB, MM10 and Rogosa agar plates for the culture of S. mutans, S. sanguinis and Lactobacillus spp., respectively. Bacterial counts were obtained by microscopic observation (10x) of the colonies and expressed in CFU/mL. Results: Bacterial counts of S. mutans and Lactobacillus spp. did not reveal age-related differences (p>0.05). Adults showed higher S. sanguinis counts than older adults with 3.7 x 105 +/- 3.8 x 105 CFU/mL and 5.9 x 104 +/- 9.4 x 104 CFU/mL, respectively (p<0.05). Conclusions: Age does not seem to affect the levels of bacterial species traditionally associated with caries. The results suggest that age may be related to colonization patterns of S. sanguinis in the oral biofilm.

Atividade antimicrobiana, antiaderente e antifúngica in vitro de plantas medicinais brasileiras sobre microrganismos do biofilme dental e cepas do gênero Candida / In vitro antimicrobial, antiadherent and antifungal activity of Brazilian medicinal plants on oral biofilm microorganisms and strains of the genus Candida

Avaliou-se in vitro a atividade antimicrobiana, antifúngica e antiaderente da aroeira-do-sertão, malva e goiabeira sobre microrganismos do biofilme dental e candidose oral. Os extratos mostraram-se eficazes, inibindo o crescimento das bactérias do biofilme dental e fungos da candidose oral, sugerindo a utilização dessas plantas como meio alternativo na terapêutica odontológica.

The antimicrobial, antifungal and antiadherent activity of aroeira-do-sertão, mallow and guava tree on oral biofilm microorganisms and oral candidiasis was evaluated in vitro. The extracts were shown to be effective in inhibiting the growth of bacteria of the oral biofilm and fungi of oral candidiasis, thus suggesting that these extracts can be used as alternative means of dental therapy.

A presença de patógenos respiratórios no biofilme bucal de pacientes com pneumonia nosocomial / Presence of respiratory pathogens in the oral biofilm of patients with nosocomial pneumonia

JUSTIFICATIVA E OBJETIVOS: O paciente internado em unidade de terapia intensiva (UTI), geralmente apresenta higiene bucal insatisfatória, podendo a região orofaríngea ser colonizada por patógenos envolvidos em pneumonia nosocomial. O objetivo deste estudo foi investigar a presença de patógenos respiratórios na cavidade bucal em pacientes em UTI. MÉTODO: Foram incluídos neste estudo transversal, 30 pacientes residentes no município de Nova Friburgo no estado do Rio de Janeiro, com idade entre 18 e 82 anos e média ponderada de 53,53 anos, sendo 17 homens e 13 mulheres, internados na UTI geral, excetuando a unidade coronariana, do Hospital Municipal Raul Sertã, Nova Friburgo, com diagnóstico de pneumonia nosocomial (PN). Foi realizada cultura das amostras do aspirado traqueal para identificar os micro-organismos responsáveis pela PN. Em contrapartida amostras microbiológicas da placa dental supragengival, da língua e do tubo do umidificador, foram analisadas para avaliação da presença do agente etiológico da PN. RESULTADOS: As bactérias mais freqüentemente encontradas no aspirado traqueal dos pacientes foram S. pneumoniae 23,3 por cento (7), P. aeruginosa 20 por cento (6), S. aureus 13,3 por cento (4), Kleibsella pneumoniae 13,3 por cento (4), Candida albicans 6,6 por cento (2), Streptococcus a-hemolítico 6,6 por cento (2), Staphylococcus sp. 6,6 por cento (2), Acinetobacter calcoaceticus - baumanii complex (A. calcoaceticus) em 1 paciente (3,3 por cento do total de pacientes), Eschericia coli (E.coli) 3,3 por cento (1), Enterobacter cloacae (E. cloacae) 3,3 por cento (1). Nesses pacientes, 70 por cento destas bactérias foram encontradas no biofilme dental, 63,33 por cento em amostras da língua, 73,33 por cento nas amostras do tubo do respirador artificial e em 43,33 por cento em todos as áreas simultaneamente. Não foram observadas diferenças significativas nas proporções das amostras dos locais de coleta (p > 0,05). CONCLUSÕES: Os resultados...

BACKGROUND AND OBJECTIVES: Hospitalized patients receiving treatment at intensive care units (ICU) usually show poor oral hygiene, and may have the mouth and oropharingeal region colonized by pathogens involved in nosocomial pneumonia. The presence of these pathogens may increase the risk for respiratory diseases. The aim of this study was to investigate the presence of respiratory pathogens in the oral cavity of hospitalized patients at ICU. METHODS: Were included in the study 30 patients from Hospital Raul Sertã, Nova Friburgo, with the diagnostic of nosocomial pneumonia, and tracheal aspirate samples were cultured to identify the causing microorganisms. In addition, microbiological samples from supragingival dental plaque, tongue and respiratory tube were cultured for the presence of a panel of respiratory pathogens. RESULTS: The most frequently found bacteria in the tracheal aspirate were S. Pneumoniae 23.3 percent (7), P. aeruginosa 20 percent (6), S. aureus 13.3 percent (4), K. pneumoniae 13.3 percent (4), C. albicans 6.6 percent (2), a-hemolytic streptococcus 6.6 percent (2), Staphylococcus sp. 6.6 percent (2), A. calcoaceticus 3.3 percent (1), E. coli 3.3 percent (1) and E. cloacae 3.3 percent (1). 70 percent (21) of these microorganisms were found in the dental biofilm, 63.33 percent (19) in tongue samples; 73.33 percent (22) in the respiratory tube; and 43.33 percent (13) in all sampling sites simultaneously. No differences in proportions could be observed between the sampling sites (p > 0.05) CONCLUSIONS: The results of this study show that respiratory pathogens associated with nosocomial pneumonia are present in the oral biofilm of hospitalized patients in ICU, which may serve as a reservoir for these microorganisms.