Background. Endodontic infections caused by remaining biofilm following disinfection with chemical fluids encourage secondary bacterial infection; hence, employing laser pulses to activate the fluids is advised to improve microbial biofilm clearance. This study investigated the performance of Er,Cr:YSGG laser in photon-induced photoacoustic streaming (PIPS) agitation of 5.25% sodium hypochlorite (NaOCl) to enhance the removal of mature Enterococcus faecalis (E. faecalis) biofilms in complex root canal systems. Methods. The mesial roots of the lower first and second molars were separated and inoculated with E. faecalis bacteria for 30 days. The roots were irrigated with 5.25% NaOCl, some of them were agitated with passive ultrasonic irrigation (PUI), and the other roots were agitated by Er,Cr:YSGG laser using PIPS at 60 µs/pulse, 5 Hz, and 0.25, 0.5, 0.75, 1, and 1.25 W. An atomic force microscope (AFM) was used as a new method to obtain the results in the isthmus area; the results that have been obtained from each group were compared with each other. ANOVA was utilized to compare the means of the test groups. Results. Based on the AFM and SEM analyses, laser agitation and passive ultrasonic activation groups have shown higher antimicrobial efficacy than the conventional syringe irrigation group (P<0.05). Conclusion. Based on the findings of this investigation, the agitation of 5.25% NaOCl solution by Er,Cr:YSGG laser in PIPS at (60 µs/pulse, 5 Hz, 1.25 W) offers better mature bacterial biofilm removal in the mesial root of lower human molars than the same irrigant with syringe irrigation and passive ultrasonic activation technique.
