RESUMO
AIM: To investigate the impact of three irrigation protocols, activated by three different methods, on mature biofilms of Enterococcus faecalis in vitro. METHODOLOGY: Root canals in 280 single-rooted teeth were instrumented using a rotary Ni-Ti system. Biofilms of E. faecalis were generated based on a previously established protocol. Samples were randomly divided into three experimental (n = 80) and one control (n = 40) group based on the irrigation protocol employed: group 1 (NaOCl + Etidronic acid), 1 : 1 mixture of 6% NaOCl and 18% etidronic acid; group 2 (NaOCl-EDTA), 3% NaOCl followed by 17% EDTA; group 3 (NaOCl-EDTA-NaOCl), 3% NaOCl followed by 17% EDTA and a final flush of 3% NaOCl. Saline served as the control. Samples were further divided into four subgroups (n = 20) based on the activation method: subgroup A, no activation; subgroup B, ultrasonic activation; group C, diode laser; group D, Er:YAG laser. Confocal laser scanning microscopy was used to assess bacterial viability in situ. Root dentine powder was obtained for determining the colony-forming units (CFU mL(-1) ). Data were analysed by appropriate statistical analyses with P = 0.05. RESULTS: All experimental irrigation protocols caused complete destruction of the biofilm in the root canal lumen. Within the dentinal tubules, all groups had a significantly higher percentage of dead bacteria than the saline control (P < 0.05). There was no significant difference between NaOCl + etidronic acid and NaOCl-EDTA-NaOCl (P > 0.05), whereas both groups brought about more bacterial reduction than NaOCl-EDTA (P < 0.05). There was no significant difference between diode laser and Er:YAG laser in any of the groups (P > 0.05). Both diode and Er:YAG laser were more effective than ultrasonic activation and conventional syringe irrigation in reducing E. fecalis biofilms (P < 0.05). CONCLUSIONS: The use of NaOCl after or in combination with a chelator caused the greatest reduction of E. faecalis. Diode laser and Er:YAG laser activation were superior to ultrasonics in dentinal tubule disinfection.