In vitro evaluation of the effectiveness of alkaline peroxide solutions in reducing the viability of specific biofilms.

Purpose: To evaluate in vitro the effectiveness of eight different cleansers containing alkaline peroxide against yeast [Candida albicans (Ca) and Candida glabrata (Cg) ], Gram-negative bacteria [Escherichia coli (Ec) and Pseudomonas aeruginosa (Pa) ], and Gram-positive bacteria [ Bacillus subtilis (Bs), Enterococcus faecalis (Ef), Staphylococcus aureus (Sa), and Streptococcus mutans (Sm) ].

Methods: Denture base acrylic resin specimens (Lucitone 550; 1264 denture base acrylic resin specimens) were obtained from a circular metal matrix (15 x 3 mm) and sterilized with microwave irradiation (650 W, 6 minutes). The specimens were contaminated with 10⁶ CFU/mL suspension of yeasts and 10⁸ CFU/mL suspensions of Gram-positive and -negative bacteria. The contaminated specimens were incubated (37°C/48 hours) and immersed (short cycle) in a stainless steel basket placed in a beaker. The beaker contained one of the following cleanser solutions, which were prepared and used according to the manufacturers' instructions (n=10): PBS (positive control, C), Fixodent (FI), Medical Interporous (MI), Kukident (KU), Efferdent Plus (EF), Equate (EQ), Kroger (KR), Kirari (KI), and Corega Tabs (CT). Reduction in the viability of microorganisms was evaluated through counting of Colony Forming Units (CFU/mL), metabolic activity (XTT), and cell viability by epifluorescence microscopy (EM) analyses. For counting CFU, the specimens (n=10 of each group) were washed and immersed in Letheen medium, from which aliquots were obtained and seeded into selective solid culture media. After incubation (37°C, 24 hours), the colonies were counted (CFU/mL). The metabolic activity of microorganisms was measured using XTT reduction assay (n=5). For EM analysis, the specimens (n= 2 of each group) were stained (Live/Dead BacLight), and the images were analyzed in terms of biofilm areas (total and living cells). Data were processed and analyzed by Wilcoxon, Mann Whitney and Kruskal-Wallis tests followed by Dunn test (α= 0.05).

Results: Significant differences were found between the experimental groups and C for the evaluated microorganisms. MI was the most effective for yeasts; this cleanser reduced the metabolic activity, viability, and total counts of both yeasts evaluated. All cleansers reduced the total counts of Gram-negative microorganisms, except CT for Pa. For Gram-negative bacteria, KR, CT, EF, and EQ significantly reduced the metabolic activity of Ec but did not remove all dead cells. FI and MI significantly reduced the metabolic activity of Pa, and the latter also removed the dead cells of Pa. FI and MI showed improved results against all Gram-positive bacteria. Both cleansers significantly decreased the metabolic activity of all bacterial species and reduced the viability of Sa, Ef, and Bs. MI also reduced the biofilm aggregates of Sm.

Clinical Significance: This study concluded that under the experimental conditions tested, denture cleansers can be used as coadjutants in denture cleaning. The evaluated cleansers exhibited disinfectant action, thereby preventing the proliferation of microorganisms on the denture surface and reducing the risk of opportunistic infections.