Photodynamic inactivation of a multispecies biofilm using Photodithazine(®) and LED light after one and three successive applications

Abstract

In this investigation, the effectiveness of successive applications of antimicrobial photodynamic inactivation (API) mediated by Photodithazine(®) (PDZ) and LED light was evaluated against a multispecies biofilm formed by Candida albicans, Candida glabrata, and Streptococcus mutans on denture base acrylic resin. Standard cell suspensions (bacteria and yeast) were inoculated on acrylic resin samples, and the biofilm was grown for 48 h (37 °C/75 rpm). API was performed by the administration of PDZ (175 and 200 mg/L) and exposure to 37.5 J/cm(2) of LED light (660 nm). Additional samples were treated with PDZ or LED light only. Untreated control samples were not submitted to light or PDZ. The conditions described were applied once or in three consecutive applications for all groups. Cell viability was determined by colony counts (CFU/mL), metabolic activity, total biomass, and confocal laser scanning microscopy (CLSM). Data were analyzed by a nonparametric two-way ANOVA and Tukey tests (α = 0.05). The results obtained demonstrated a significant effect (p < 0.05) of number of applications and treatment groups for CFU/mL, and S. mutans showed the highest susceptibility to API. The metabolic activity of the multispecies biofilm was significantly reduced (p < 0.05) after API for both numbers of applications, which were also significantly different (p < 0.05) between them. The total biomass of the biofilm was significantly different (p < 0.05) only between groups submitted to one and three API applications. CLSM showed a visual increase of dead cells after API. API-mediated PDZ was effective in reducing the cell viability of multispecies biofilm. Three consecutive applications of API were more effective for reducing the cell viability and the total biomass of multispecies biofilm.