The in vivo antimicrobial activity of 0.12% and 0.2% chlorhexidine (CHX) on the salivary flora up to 7 h after its application, using epifluorescence microscopy with the SYTO 9/propidium iodide dual staining, was evaluated. Fifteen volunteers performed a single mouthrinse with sterile water (SM-water), a single mouthrinse with 0.12% CHX (0.12% SM-CHX) and a single and double mouthrinse with 0.2% CHX (0.2% SM-CHX and 0.2% DM-CHX). Samples of saliva were taken at 30 s, and 1, 3, 5, and 7 h after each application. In comparison with SM-water, 0.2% CHX (SM and DM) showed a significant antibacterial effect up to 7 h after the mouthrinse, whereas this effect only persisted up to 5 h after the 0.12% SM-CHX mouthrinse. On comparing the two concentrations of CHX, significantly higher percentages of bacterial vitality were observed in all the saliva samples after the use of 0.12% CHX than after 0.2% CHX. On comparison of the 0.2% SM-CHX and 0.2% DM-CHX, significantly higher percentages of live bacteria were observed in the saliva samples taken at 1, 3, 5, and 7 h after the single mouthrinse compared with the double mouthrinse. The 0.2% CHX mouthrinse had the greatest antimicrobial activity on the salivary flora up to 7 h after its application, with a progressive recovery in bacterial vitality. The differences observed with respect to the 0.12% CHX mouthrinse demonstrate the influence of the concentration on its immediate antimicrobial activity and substantivity.
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.
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