Amphotericin B, fluconazole, and nystatin as development inhibitors of Candida albicans biofilms on a dental prosthesis reline material: Analytical models in vitro

Abstract

Statement of problem: The use of antifungals has been suggested during the treatment of denture stomatitis associated with Candida albicans biofilms. However, how time, material surface, and substrates present during adhesion and biofilm development can influence clinical treatment is unclear.

Purpose: The purpose of this in vitro study was to investigate the growth kinetics of C. albicans biofilms on surfaces of specimens under the influence of adsorbed films and to evaluate the antibiofilm efficacy of antifungal agents: amphotericin B, fluconazole, and nystatin.

Material and methods: Specimens of Silagum-Comfort Soft Relining were submerged in preconditioning systems: phosphate-buffered saline, artificial saliva, fetal bovine serum, and artificial saliva+fetal bovine serum. Planktonic cells were incubated (phosphate-buffered saline+specimens) for 1.5 hours (adhesion phase) and washed with phosphate-buffered saline solution. The specimens were then incubated (YNB+glucose) for 8, 24, and 48 hours (initial, intermediate, and maturation phases). The biofilm sessile minimum inhibitory concentration was determined by the broth microdilution method (7.81 to 500 μg/mL). The metabolic activity of the biofilms was tested by colorimetric assay (cell metabolic activity). Cell viability, relative biomass (μm³), and the thickness of the biofilm (μm) were evaluated by confocal laser scanning microscopy.

Results: The highest bioactivity was recorded in the presence of fetal bovine serum. Biofilms treated with fluconazole and amphotericin B were partially inhibited in a dose-dependent manner. Nystatin inhibited metabolic activity mainly from ≥15.63 or 62.5 μg/mL. Variations in magnitude parameters (relative biomass and thickness) were observed depending on the development phases of biofilms, whereas biological parameters (percentage of nonviable cells) were constant throughout the formation of C. albicans biofilms.

Conclusions: The data suggest that partial (fluconazole and amphotericin B) or more effective (nystatin) reduction of metabolic activity of C. albicans biofilms occurred depending on the time and the antifungal and its concentrations.