Brazilian red propolis reduces orange-complex periodontopathogens growing in multispecies biofilms.

This study investigated the antimicrobial effects of the ethanolic extract of Brazilian red propolis (BRP) on multispecies biofilms. A seven-day-old subgingival biofilm with 32 species was grown in a Calgary device. Biofilms were treated with BRP (1,600, 800, 400 and 200 µg ml-1) twice a day for 1 min, starting from day 3. Chlorhexidine (0.12%) and dilution-vehicle were used as positive and negative controls, respectively. On day 7, metabolic activity and the microbial composition of the biofilms by DNA-DNA hybridization were determined. The viability data were analyzed by one-way ANOVA followed by Tukey's post hoc, whereas the microbial composition data were transformed via BOX-COX and analyzed using Dunnett's post hoc. BRP (1,600 µg ml-1) decreased biofilm metabolic activity by 45%, with no significant difference from chlorhexidine-treated samples. BRP (1,600 µg ml-1) and chlorhexidine significantly reduced levels of 14 bacterial species compared to the vehicle control. Taken together, BRP showed promising antimicrobial properties which may be useful in periodontal disease control.

Brazilian Red Propolis Is as Effective as Amoxicillin in Controlling Red-Complex of Multispecies Subgingival Mature Biofilm In Vitro.

This study investigated the effects of Brazilian Red Propolis (BRP) extract on seven-day-old multispecies subgingival biofilms. Mixed biofilm cultures containing 31 species associated with periodontal health or disease were grown for six days on a Calgary device. Then, mature biofilms were treated for 24 h with BRP extract at different concentrations (200-1600 µg/mL), amoxicillin (AMOXI) at 54 µg/mL (positive control) or vehicle (negative control). Biofilm metabolic activity was determined by colorimetry, and bacterial counts/proportions were determined by DNA-DNA hybridization. Data were analyzed by Kruskal-Wallis and Dunn's tests. Treatment with BRP at 1600, 800 and 400 µg/mL reduced biofilm metabolic activity by 56%, 56% and 57%, respectively, as compared to 65% reduction obtained with AMOXI. Mean total cell counts were significantly reduced in all test groups (~50-55%). Lower proportions of red, green and yellow complex species were observed upon treatment with BRP (400 µg/mL) and AMOXI, but only AMOXI reduced the proportions of Actinomyces species. In conclusion, BRP extract was as effective as AMOXI in killing seven-day-old multispecies biofilm pathogens and did not affect the levels of the host-compatible Actinomyces species. These data suggest that BRP may be an alternative to AMOXI as an adjunct in periodontal therapy. In vivo studies are needed to validate these results.

In Vitro Antimicrobial Effect of Cetylpyridinium Chloride on Complex Multispecies Subgingival Biofilm.

Periodontopathogenic subgingival biofilm is the main etiological agent of periodontitis. Thus, a search for antimicrobials as adjuvant for periodontal treatment in the literature is intense. Cetylpyridinium chloride (CPC) is a well-known antimicrobial agent commonly used in mouthrinses. However, CPC effects on a complex biofilm model were not found over the literature. Therefore, the aim of this manuscript is to evaluate 0.075% CPC antimicrobial properties in a multispecies subgingival biofilm model in vitro. The subgingival biofilm composed by 31 species related to periodontitis was formed for 7 days, using the calgary device. The treatments with CPC and chlorhexidine (CHX) 0.12% (as positive control) were performed 2x/day, for 1 min, from day 3 until the end of experimental period, totaling 8 treatments. After 7 days of biofilm formation, biofilm metabolic activity was evaluated by a colorimetric reaction and biofilms microbial composition by DNA-DNA hybridization. Statistical analysis was performed using ANOVA with data transformed via BOX-COX followed by Dunnett post-hoc. Both CPC and CHX reduced biofilm metabolic activity in 60% and presented antimicrobial activity against 13 different species. Specifically, only CHX reduced levels of F.n. vicentii and P. gingivalis while only CPC reduced A. odontolyticus and A. israelli. CPC was as effective as CHX as antimicrobial through in vitro complex multispecies subgingival biofilm. However, future studies using in vivo models of experimental periodontal disease should be performed to prove such effect.

In vitro antimicrobial effect of cetylpyridinium chloride on complex multispecies subgingival biofilm

Abstract Periodontopathogenic subgingival biofilm is the main etiological agent of periodontitis. Thus, a search for antimicrobials as adjuvant for periodontal treatment in the literature is intense. Cetylpyridinium chloride (CPC) is a well-known antimicrobial agent commonly used in mouthrinses. However, CPC effects on a complex biofilm model were not found over the literature. Therefore, the aim of this manuscript is to evaluate 0.075% CPC antimicrobial properties in a multispecies subgingival biofilm model in vitro. The subgingival biofilm composed by 31 species related to periodontitis was formed for 7 days, using the calgary device. The treatments with CPC and chlorhexidine (CHX) 0.12% (as positive control) were performed 2x/day, for 1 min, from day 3 until the end of experimental period, totaling 8 treatments. After 7 days of biofilm formation, biofilm metabolic activity was evaluated by a colorimetric reaction and biofilms microbial composition by DNA-DNA hybridization. Statistical analysis was performed using ANOVA with data transformed via BOX-COX followed by Dunnett post-hoc. Both CPC and CHX reduced biofilm metabolic activity in 60% and presented antimicrobial activity against 13 different species. Specifically, only CHX reduced levels of F.n. vicentii and P. gingivalis while only CPC reduced A. odontolyticus and A. israelli. CPC was as effective as CHX as antimicrobial through in vitro complex multispecies subgingival biofilm. However, future studies using in vivo models of experimental periodontal disease should be performed to prove such effect.

Resumo O biofilme subgengival periodontopatogênico é o principal agente etiológico da periodontite. Assim, a pesquisa de antimicrobianos como adjuvantes para o tratamento periodontal na literatura é intensa. Cloreto de cetilpiridínio (CPC) é um agente antimicrobiano comumente usado em enxaguatórios bucais. No entanto não foram encontrados na literatura estudos avaliando os efeitos do CPC em um modelo complexo de biofilme. Portanto, o objetivo deste artigo é avaliar as propriedades antimicrobianas do cloreto de cetilpiridinio 0,075% em um modelo de biofilme subgengival multiespécie in vitro. O biofilme subgengival composto por 31 espécies relacionadas à periodontite foi formado por 7 dias, utilizando o dispositivo calgary. Os tratamentos com CPC e clorexidina (CHX) 0,12% (controle positivo) foram realizados 2x/dia, por 1 min, do dia 3 até o final do período experimental, totalizando 8 tratamentos. Após 7 dias de formação do biofilme, a atividade metabólica do biofilme foi avaliada por reação colorimétrica e a composição microbiana dos biofilmes por hibridização DNA-DNA. A análise estatística foi realizada usando ANOVA com dados transformados via BOX-COX seguido do teste de Dunnett. Tanto o CPC como a CHX reduziram a atividade metabólica do biofilme em aproximadamente 60% e apresentaram atividade antimicrobiana contra 13 espécies diferentes. Especificamente, apenas os níveis de F.n. Vicentii e P. gingivalis foram reduzidos somente pelo tratamento com a CHX enquanto apenas o CPC reduziu A. odontolyticus e A. israelli. O CPC foi tão eficaz quanto o CHX como antimicrobiano através de biofilme subgengival complexo multiespecífico in vitro. No entanto, futuros estudos usando modelos in vivo de doença periodontal experimental devem ser realizados para comprovar tal efeito.