The antimicrobial effect of different ozone protocols applied in severe curved canals contaminated with Enterococcus faecalis: ex vivo study

The aim of this study was to evaluate 3 different irrigation protocols using ozone on monoculture biofilm of Enterococcus faecalis. Fifty disto-buccal roots of maxillary first molars were instrumented up to the size 25.08, sterilized and then incubated with Enterococcus faecalis for 21 days for biofilm maturation in the root canal walls. After contamination, the initial collection (S1) was performed and the 50 samples were divided in 3 experimental groups (n = 15) and 1 positive control group (n = 5) as follows: Group1: Ozonated water (40 mu g/ml); Group2: gaseous ozone (40 mu g/ml); Group3: Ozonated water (40 mu g/ml) under continuous ultrasonic irrigation (CUI) and, Group 4: 2.5% sodium hypochlorite (positive control). The final collection (S2) was performed using hedstroem files # 25 by scraping the root canal wall and absorbed paper cone. The collected samples were analyzed by CFU/ml count and qPCR. To compare the effect of irrigation methods and to investigate E. faecalis count data, Kruskal-Wallis and Wilcoxon nonparametric tests were used at p < 05. The values of the differences between the S1 values and the S2 values revealed that the groups did not differ significantly for either E. faecalis CFU count data (p = 0.713) or q-PCR (p = 0.185), however, it is observed that for all groups, the CFU count of E. faecalis decreased over 99%. For the q-PCR assay, the reduction was on the order of 74-95%. Only the NaOCl positive control group presented total elimination of CFU/mL bacterial counting. The ozone protocols assessed were similar in regard to Enterococcus faecalis reduction.