title: Validation of ATP bioluminescence as a tool to assess antimicrobial effects  of mouthrinses in an in vitro subgingival-biofilm model.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: ETEP (Aetiology and Therapeutics of Periodontal Diseases) Research Group, University Complutense of Madrid, Spain.
authors.forename: María-Carmen
authors.initials: MC
authors.lastname: Sánchez
authors.forename: Arancha
authors.initials: A
authors.lastname: Llama-Palacios
authors.forename: María-José
authors.initials: MJ
authors.lastname: Marín
authors.forename: Elena
authors.initials: E
authors.lastname: Figuero
authors.forename: Rubén
authors.initials: R
authors.lastname: León
authors.forename: Vanessa
authors.initials: V
authors.lastname: Blanc
authors.forename: David
authors.initials: D
authors.lastname: Herrera
authors.forename: Mariano
authors.initials: M
authors.lastname: Sanz
mesh_terms: D000255:Adenosine Triphosphate; D000891:Anti-Infective Agents, Local / Q000494:pharmacology*; D018407:Bacterial Physiological Phenomena / Q000187:drug effects*; D018441:Biofilms / Q000187:drug effects*; D008163:Luminescent Measurements; D009067:Mouthwashes / Q000494:pharmacology*
chemical_list: D000891:Anti-Infective Agents, Local; D009067:Mouthwashes; D000255:Adenosine Triphosphate