Cross-kingdom interaction of Candida albicans and Actinomyces viscosus elevated cariogenic virulence

Objective: How the interactions between Candida albicans and Actinomyces viscosus contributed to the root caries was not clear. This study aimed to investigate their cross-kingdom interactions on the biomass and the cariogenic virulence in dual-species biofilms. Design: Suspensions of C. albicans and A. viscosus were formed the mono and polymicrobial biofilms in vitro. Crystal violet assay, viable plate count, scanning electron microscopy and fluorescence in situ hybridization were used to analyze the biomass and biofilm structure. Glycolytic pH drop and the spectrophotometric method were used to evaluate the acid production and hydroxyapatite dissolution, respectively. The exopolysaccharide production was measured by the anthrone-sulfuric acid method, while the adhesion force was measured by atomic force microscopy. Results: The biomass and colony-forming units of mixed-species were significantly increased compared to that of the mono-species at 24 h, 48 h, 72 h. The structure of dual-species biofilm had more microcolonies and was much denser. The dual-species biofilms significantly decreased the pH value and damaged the hydroxyapatite compared with the mono-species biofilms at various time points, indicating the strong cariogenic virulence. Moreover, the dual-species biofilms significantly enhanced the exopolysaccharide production and adhesion force suggesting the increase of biofilm adhesion. Conclusions: Cross-kingdom interactions of C. albicans and A. viscosus significantly elevated the biomass and cariogenic virulence of dual-species biofilm.