Experimental human root canal irrigant NaOCl against Enterococcus faecalis and 3T3, and determination of cytotoxicity effect

Abstract

The main aim of endodontic treatment is disinfection of root canal and to prevent chances of reinfection. The most commonly isolated species due to oral infections is Enterococcus faecalis. For nonsurgical endodontic procedures Sodium hypochlorite (NaOCl) has been the irrigant of choice. The mechanism by which endodontic irrigants induce cytotoxicity is still unclear. However, many studies clearly indicated that rapid expression of the reactive oxygen species (ROS) leads to free radicals formation which results in cytotoxicity and cell death. Hence this study was done to determine the viability of cells and oxidative stress mediated by NaOCl, an endodontic irrigant. The irrigants were tested for their effect against fibroblast isolated from human primary buccal mucosa and against 3T3 Cell line. Antibacterial activity was performed against Enterococcus faecalis. Cytotoxicity was determined by MTT. To determine the oxidative stress, total intracellular glutathione, superoxide radical scavenging activity, and catalase assays were performed. The MIC (Minimal Inhibitory Concentration) for the irrigantsagainst Enterococcusfaecalis was found to be 10 µl. 10 µl of NaOCl plain 5.2% produced the same effect as that of 10 ìl of NaOCl plain 3%. The higher concentration of the irrigants decreased viability of the cells during dye exclusion assay. Enzyme based study showed there is a decrease in enzyme dehydrogenase when treat with irrigants. Glutathione, SOD level was increased gradually on 3T3 cells. But CAT level was increased when the irrigants concentration less. The results of this study indicated that endodontic irrigants were potentially controlling the Enterococcus faecalis and non-toxic/reduced viability of 3T3 cells by MTT which could be due to the oxidative stress and loss of cellular integrity probably due to the liberation of ROS evidenced by the alteration of antioxidant enzymes Glutathione, SOD and CAT.