Abstract
Objectives. The aim of this investigation was to evaluate whether the adenosine triphosphate (ATP) bioluminescence method is an appropriate tool to assess the efficacy of antiseptic mouthrinses in terms of quantitative reductions of total viable microbial counts in mixed biofilm populations in vitro. Study Design. Three mouthrinses, containing respectively, chlorhexidine and cetylpyridinium chloride (CHX/CPC), essential oils (EO) and amine fluoride/stannous fluoride (AFSF), as well as Phosphate Buffered Saline (PBS) used as control, were tested in an in vitro static biofilm model by ATP bioluminescence and compared to culture method. Biofilms were grown on saliva-coated hydroxyapatite disks for 72 hours and then exposed for 1 minute to the mouthrinse or control by immersion. The antibacterial effect of the rinses was tested by analysis of variance. The reliability of the ATP bioluminescence method was assessed by calculating the Pearson correlation coefficients when compared to the viable cell counts obtained by culture. Results. Using ATP bioluminescence, the antimicrobial activity of the tested mouthrinses was demonstrated when compared to the PBS control. The ATP bioluminescence values were significantly correlated (0.769, p<0.001) to the viable cell counts. CHX/CPC and AFSF showed similar antimicrobial activity, although AFSF had a less homogeneous effect, being both more effective than the EO rinse. Conclusion. ATP bioluminescence viability testing may be considered a useful tool to assess the in vitro efficacy of antibacterial compounds. In the proposed model, CHX/CPC and AFSF containing mouthrinses demonstrated superior antimicrobial activity, as compared to EO rinses, in a multispecies biofilm model. Key words:Biofilm, ATP bioluminescence,mouthrinse, essential oils, chlorhexidine, amine fluoride/stannous fluoride.
Highlights
The effective control of the dental plaque is the key in the prevention of periodontal diseases
The objective of this study was to validate the use of the adenosine triphosphate (ATP) bioluminescence method for rapid quantitative evaluation of viable total oral bacteria in biofilm samples after exposure to the antimicrobial agents
The obtained results have shown that the ATP bioluminescence method was capable of quantifying bacterial vitality, when applied to a validated in vitro biofilm model
Summary
The effective control of the dental plaque is the key in the prevention of periodontal diseases. Our research group has recently developed and tested such an in vitro biofilm model reporting its structure, viability and bacterial kinetics [20] This model uses six bacteria from the subgingival biofilm, containing initial (Streptococcus oralis and Actinomyces naeslundii), early (Veillonella parvula), secondary (Fusobacterium nucleatum), and late colonizers (Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans). It is, the main purpose of this investigation evaluate the possibility to use ATP bioluminescence method for rapid quantitative evaluation of viable total oral bacteria in biofilms samples, as compared to standard culture methods, to test the bactericidal efficacy of antiseptic mouthrinses. We used three commercially available antiseptic mouthrinses containing respectively, chlorhexidine digluconate (CHX) and cetyl-pyridinium chloride (CPC), essential oils (EO), and amine fluoride/stannous fluoride (AMSF) in a tested and validated in vitro biofilm model [20]
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