Evaluation of chlorhexidine substantivity on salivary flora by epifluorescence microscopy

OBJECTIVE: To evaluate the in vivo antimicrobial activity of chlorhexidine (CHX) in saliva 7 h after its application using an epifluorescence microscopy technique. SUBJECTS AND METHODS: Fifteen volunteers performed a single mouthrinse with sterile water (SM-water) and with 0.2% CHX (SM-0.2% CHX). Saliva samples were taken at 30 s and 1, 3, 5 and 7 h after each application. The bacterial suspension was mixed with the SYTO 9/propidium iodide staining and observed using an Olympus BX51 microscope. The mean percentage of viable bacteria was calculated for each sample. RESULTS: In comparison with baseline values, the frequency of viable bacteria decreased significantly at 30 s after the SM-0.2% CHX (P < 0.001) and presented significant antibacterial activity up to 7 h after the mouthrinse (P < 0.001). In comparison with SM-water, the prevalence of viable bacteria was significantly lower at 30 s after the SM-0.2% CHX (P < 0.001) and showed a significant antibacterial effect up to 7 h after the mouthrinse (P < 0.001). CONCLUSIONS: Epifluorescence microscopy permits evaluating the antimicrobial activity of CHX on the salivary flora in real-time. Fluorescence assays could be particularly useful to analyse simultaneously the effect of antimicrobials that alter the cytoplasmic membrane integrity on different oral ecosystems.