Interkingdom interaction between C. albicans and S. salivarius on titanium surfaces

In oral candidiasis models, Candida albicans and Streptococcus salivarius sp. biofilms have an antagonistic relationship. Due to this, S. salivarius have been used experimentally as probiotic. However, the interaction between these microorganisms in the peri-implantitis-like microenvironment remains unknown. This study aimed to evaluate the interaction between C. albicans and S. salivarius biofilms developed on titanium surfaces, under reduced oxygen levels.

Titanium specimens were pre-conditioned with artificial saliva (1 h, 37 °C). Single-species biofilms of C. albicans (ATCC 90028) and co-culture biofilms of C. albicans and S. salivarius (ATCC 7073) was developed for 24 and 72 h on titanium specimens. Subsequently, the effect of these intervals of biofilm formation and the interactions among the cells were evaluated. Biofilms from cultures were collected and analyzed for cell viability (CFU/mL), biofilm biomass, and total protein dosage. Data were analyzed using Mann-Whitney test (α = 5%). In addition, co-culture biofilms were analyzed using fluorescence microscopy.

C. albicans growth did not change due to the presence of S. salivarius. Besides, co-culture biofilms showed a significant difference in the number of viable cells between 24 and 72 h of biofilm development (p < 0.05). The highest biofilm biomass and protein dosage were observed in co-cultures at 72 h of biofilm development. Fluorescence microscopy showed that co-cultures biofilms at 24 h have limited number of pseudo-hyphal and hyphae cells of C. albicans. At 72 h, these types of cells have increased. S. salivarius in both stages of development was present in some clusters surrounded by C. albicans.

Co-cultivation of C. albicans with S. salivarius in biofilms developed on titanium surfaces, under lower oxygen levels, did not affect fungus growth. In addition, S. salivarius did not hind C. albicans virulence. These findings suggest that the use of S. salivarius as a probiotic would be ineffective in peri-implant disease treatment.