Incorporation of zinc into cetylpyridinium chloride mouthwash affects the composition of multispecies biofilms.

This study evaluated the effect of a mouthwash containing 0.075% cetylpyridinium chloride and 0.28% zinc lactate (CPC + Zn) in a multispecies biofilm model. A 7-days 33-species biofilm, formed on Calgary device, was 1-min treated with: 0.12% chlorhexidine (CHX), culture medium (negative control), 0.075% cetylpyridinium chloride (CPC) or CPC + Zn, 2x/day, from day 3 until day 6. The metabolic activity and the microbial composition were evaluated by colorimetric method and checkerboard DNA-DNA hybridization, respectively. The three antimicrobials (CPC, CPC + Zn and CHX) reduced metabolic activity, total biofilm count and several species counts, including Porphyromonas gingivalis, Fusobacterium nucleatum, Parvimonas micra, Campylobacter gracilis and Streptococcus mutans. However, only CPC + Zn reduced counts of the pathogen Prevotella intermedia and did not interfere with the levels of some beneficial species in relation to the negative control. The treatment of multispecies subgingival biofilm with CPC + Zn was effective in controlling periodontal pathogens and favored the colonization of health-associated bacterial species.

Statins and oral biofilm: Simvastatin as a promising drug to control periodontal dysbiosis.

OBJECTIVES: This study evaluated antimicrobial activity of atorvastatin, pravastatin, rosuvastatin, and simvastatin against oral bacteria, and the interaction of simvastatin with standard antimicrobials (amoxicillin and metronidazole). METHODS: Minimal inhibitory concentration assays were performed with Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinomyces odontolyticus, Streptococcus oralis, Streptococcus mitis, Streptococcus salivarius, Streptococcus sanguinis, and Streptococcus gordonii; checkerboard microdilution assays between simvastatin and standard antimicrobials; monospecies and multispecies biofilms. RESULTS: Simvastatin showed the best antimicrobial activity against most species (MIC range from 3.12 to 25 µg/ml), highlighting the sensitivity of P. gingivalis. In the checkerboard assay, synergistic interaction was found between simvastatin and amoxicillin against S. oralis and S. sanguinis. P. gingivalis biofilm was inhibited by simvastatin at 10 and 50× Minimal inhibitory concentration, with similar effects to metronidazole. For multispecies biofilm, SMV reduced the biofilm metabolic activity (79%) and total counts (87%), comparable to amoxicillin. Simvastatin also reduced bacterial counts of Veilonnella parvula, P. gingivalis, Streptococcus mutans, Actinomyces naeslundii, P. intermedia, and Capnocytophaga ochracea in the multispecies biofilm. CONCLUSIONS: Simvastatin showed antimicrobial and antibiofilm activity against oral bacteria and may contribute to the control of dysbiosis, and may be considered in clinical studies as an adjuvant in the treatment of periodontitis.

Development of a multispecies periodontal biofilm model within a stirred bioreactor.

The objective of this work was to develop a subgingival biofilm model using a stirred bioreactor. Discs of bovine teeth were adapted to a stirred bioreactor filled with a culture medium containing bacterial species associated with periodontal health or disease. After anaerobic incubation, the biofilms growing on the substratum surfaces were collected and analyzed. The mean number of Colony-forming Units (CFUs) varied, but with no difference between 3 and 7 days of biofilm formation (p > 0.05). Scanning Electron Microscopy (SEM) analysis showed a uniform biofilm layer covering the cement layer of the root surface containing bacteria with diverse morphology. In checkerboard DNA-DNA hybridization, bacterial species were identified in both biofilms. In conclusion, a subgingival biofilm model was developed using a stirred bioreactor, allowing the in vitro reproduction of complex microbial communities. This is an advanced model that may be useful to mimic complex clinical periodontal biofilms.