Antibiofilm activity of ultrasonic and diode laser activated sodium hypochlorite, chitosan, and chlorhexidine: A confocal laser scanning microscopic in vitro study

J Conserv Dent Endod. 2023 Jul-Aug;26(4):441-446. doi: 10.4103/jcd.jcd_224_23. Epub 2023 Jul 28.

Abstract

Aim: The aim of this study was to assess and compare the effect of ultrasonic and diode laser-activated sodium hypochlorite (NaOCl), chitosan, and chlorhexidine (CHX) on the removal of Enterococcus faecalis biofilm adherent to the root canal using a confocal laser scanning microscope (CLSM).

Materials and methods: Root canals in 112 single-rooted teeth were instrumented using a rotary Ni-Ti system. Biofilms of E. faecalis were generated based on an established protocol. Samples were randomly divided into three experimental (n = 28) and one control (n = 28) group based on the irrigation protocol employed and the three experimental groups were further subdivided into subgroups based on the activation protocol (subgroup A - ultrasonic activated and subgroup B - diode laser activated). The groups were Group 1 (control), Group 2 (3% NaOCl for 6 min; subgroup A - activated using a diode laser, subgroup B - ultrasonic activation), Group 3 (2% CHX for 6 min; subgroup A - activated using a diode laser, subgroup B - ultrasonic activation), and Group 4 (0.2% chitosan for 6 min; subgroup A - activated using a diode laser, subgroup B - ultrasonic activation. Confocal laser scanning microscopy was used to assess bacterial viability in situ. Data were analyzed by appropriate statistical analyses with P = 0.05.

Results: All experimental irrigation protocols destroyed 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 CHX activated with ultrasonics, CHX activated with a diode laser, chitosan activated with ultrasonics and chitosan activated with diode laser groups (P > 0.05), whereas NaOCl ultra and NaOCl diode groups brought about more bacterial reduction than other groups (P < 0.05). The mean effectiveness and the bacterial kill ability were seen highest for the NaOCl activated with the ultrasonics group. There was no significant difference between diode laser activation and ultrasonic activation in CHX activated with ultrasonics, CHX activated with a diode laser, chitosan activated with ultrasonics and chitosan activated with diode laser groups (P > 0.05), but there was a significant difference between diode laser and ultrasonic activation in NaOCl group. Ultrasonic activation of the NaOCl was more effective than diode activation in reducing E. feacalis biofilms (P < 0.05).

Conclusions: The use of NaOCl with the activation by ultrasonics caused the greatest reduction of E. faecalis. Ultrasonic activation was found superior to diode laser activation in dentinal tubule disinfection.

Keywords: Biofilm; Enterococcus faecalis; chitosan; chlorhexidine; confocal microscopy; diode laser; sodium hypochlorite; ultrasonics.