[Killing activity of nonequilibrium plasma against young and old Enterococcus faecalis biofilms with long-term exposure in infected root canals in vitro].

Objective To evaluate the antimicrobial activity of nonequilibrium plasma against Enterococcus faecalis (Ef) biofilms in vitro and to obtain novel evidence of root canal disinfection with nonequilibrium plasma. Methods Sterile cover slips and single-rooted canals were filled with Ef and incubated to form 1-week-old and 3-week-old biofilms, respectively. The infected samples were subjected to nonequilibrium plasma, 2% chlorhexidine digluconate (CHX) and saline for 3, 10 and 30 minutes, respectively. After treatment, the killing effectiveness of nonequilibrium plasma was analyzed by using laser scanning confocal microscopy (LSCM) and colony forming unit (CFU) counting. Results The 3-dimentional reconstruction LSCM images showed that about 48.3%-79.8% of 1-week-old Ef biofilm cells and 40.0%-67.4% of 3-week-old biofilm cells were killed by nonequilibrium plasma and 2% CHX compared to saline (P<0.05). The proportion of killing activity was lower after 3 minutes (40.0%-50.9% killing) than after 10 minutes (65.3%-77.8% killing) and 30 minutes (66.4%-79.8% killing) (P<0.05). And the killing of biofilm bacteria was fastest during the first 3 minutes (13.3%-17.0% killing per minute) and slow down greatly after 10 minutes. Remarkably more bacteria were killed in 1-week-old Ef biofilms (48.3%-79.8% killing) than in 3-week-old biofilms (P<0.05). Conclusions The nonequilibrium plasma killed more Ef biofilm cells in infected root canals showed promotional as an additional approach against bacterial biofilms during root canal disinfection. Key words: Biofilms; Enterococcus faecalis; Chlohexidine; Root canal disinfection; Nonequlibrium plasma