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Antibacterial activity of chlorhexidine after final irrigation with ethanol: CLSM and culture‐based method analysis

This study investigated the effect of 95% ethanol on the antibacterial properties of 2% chlorexidine (CHX) over monospecies biofilm (Enterococcus faecalis) through a culture‐based method, and over multispecies biofilm using confocal laser scanning microscopy (CLSM). For monospecies model, E. faecalis biofilm was induced in 40 root canals. The irrigation procedures were: S—saline solution; S/CHX—saline solution + CHX; E—ethanol; and E/CHX—ethanol + CHX. Microbial sampling was performed at three periods: before (S1), immediately after (S2), and 72 h after the final flush (S3). For multispecies biofilm model, 28 sterilized bovine dentin blocks were fixed on a removable orthodontic device to allow intraoral biofilm development. Seven samples were used in each group. Statistical analysis was carried out by using the Kruskal–Wallis test and Dunn's test for multiple comparisons. There was a significant reduction in CFUs count immediately after the final flush (S2) in all experimental groups (P < 0.05). However, only S/CHX, E and E/CHX groups had CFU counts close to zero, without differences among them (P > 0.05). After 72h (S3), the S/CHX and E/CHX groups had CFU counts near zero (P > 0.05). The CFU count increased in S3 for S and E groups (P < 0.05). CLSM showed that the percentages of remaining live cells were similar in S/CHX, E, and E/CHX groups (P > 0.05). The S group had the highest percentage of live cells (P < 0.05). The 95% ethanol did not interfere in the antibacterial properties of 2% CHX over mono‐ and multispecies biofilms. Microsc. Res. Tech. 78:682–687, 2015. © 2015 Wiley Periodicals, Inc.