Validation of ATP bioluminescence as a tool to assess antimicrobial effects of mouthrinses in an in vitro subgingival-biofilm model

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.

Effects of Azithromycin, Metronidazole, Amoxicillin, and Metronidazole plus Amoxicillin on anIn VitroPolymicrobial Subgingival Biofilm Model

“…The model presents several advantages over previously existing models (16)(17)(18)(19)(20)(21), in that we were able to consistently establish 35 species, compared with the previous maximum of 5 to 10 species (18,20,21). Further, the Calgary Biofilm Device afforded a high-throughput system to grow several repeats of biofilms with good consistency across pegs and plates, as evidenced by the high minimum similarity coefficient values.…”

“…The first step in such an endeavor is to demonstrate efficacy against subgingival microorganisms in vitro, ideally in models involving organized biofilms that respond like those in vivo. Currently used in vitro oral multispecies biofilm models all suffer from one or more limitations, generally comprising only up to 5 or 10 species (16)(17)(18)(19)(20)(21). In addition, existing models lack reproducibility and have low throughput (17,19).…”

In vitro method for prediction of plaque reduction by dentifrice

“…In contrast, ATP is a precise and valid metric previously applied to similar oral biofilm testing. ATP of dispersed oral biofilm cells after treatment was successfully used to quantitate relative efficacy of mouthrinses and mouthrinse ingredients (Pan et al, 2010;Sánchez et al, 2013). Sánchez et al (2013) also found that viable cell counts and ATP levels were significantly (p b 0.001) correlated.…”

Commercial antimicrobials mouthrinses on caries and periodontitis-related biofilm control: a review of literature

“…Sánchez et al [40] evaluated the antimicrobial effect of three commercial mouthrinses (0.12% CHX plus 0.05% CPC, EO, and fluoride/stannous fluoride -AFSF) on mixed periodontopathogenic biofilm (S. oralis, V. parvula, A. naeslundii, F. nucleatum, A. actinomycetemcomitans and P. gingivalis) in vitro using ATP bioluminescence and CFU methods. 72h-biofilm was exposed to mouthrinses or control for 1 min.…”