Chemical efficacy of several NaOCl concentrations on biofilms of different architecture: new insights on NaOCl working mechanisms.

ABSTRACT

AIM: To investigate the anti-biofilm efficacy and working mechanism of several NaOCl concentrations on dual-species biofilms of different architecture as well as the changes induced on the architecture of the remaining biofilms. METHODOLOGY: Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 were co-cultured under different growth conditions on saliva-coated hydroxyapatite discs. A constant-depth film fermenter (CDFF) was used to grow steady-state, four-day mature biofilms (dense architecture). Biofilms were grown under static conditions for 4 days within a confined space (less dense architecture). Twenty microlitres of buffer, 2-, 5-, and 10% NaOCl were applied statically on the biofilms for 60 s. Biofilm disruption and dissolution, as well as bubble formation, were evaluated with optical coherence tomography (OCT). The viscoelastic profile of the biofilms post-treatment was assessed with low load compression testing (LLCT). The bacteria/extracellular polysaccharide (EPS) content of the biofilms was examined through confocal laser scanning microscopy (CLSM). OCT, LLCT and CLSM data were analysed through one-way analysis of variance (ANOVA) and Tukey's HSD post-hoc test. Linear regression analysis was performed to test the correlation between bubble formation and NaOCl concentration. The level of significance was set at a < 0.05. RESULTS: The experimental hypothesis according to which enhanced biofilm disruption, dissolution and bubble formation were anticipated with increasing NaOCl concentration was generally confirmed in both biofilm types. Distinct differences between the two biofilm types were noted with regard to NaOCl anti-biofilm efficiency as well as the effect that the several NaOCl concentrations had on the viscoelasticity profile and the bacteria/EPS content. Along with the bubble generation patterns observed, these led to the formulation of a concentration and biofilm structure-dependent theory of biofilm removal. CONCLUSIONS: Biofilm architecture seems to be an additional determining factor of the penetration capacity of NaOCl, and consequently of its anti-biofilm efficiency.