In vitro cytotoxicity and antibacterial activity of hypochlorous acid antimicrobial agent

Bacteria-associated oral diseases such as dental caries and periodontitis are widespread epidemics that cause oral pain and loss of function. The purpose of this study was to evaluate the in vitro cytotoxicity and antibacterial activity of different concentrations of hypochlorous acid (HOCl).

Five different concentrations (100, 200, 300, 400, and 500 ppm) of HOCl were evaluated for their antimicrobial efficacy against Gram-negative (A. actinomycetcmcomitans and P. gingivalis) and Gram-positive bacteria (S. mutans and S. sanguinis) after treatment for 1 and 10 min. Sodium hypochlorite (NaOCl) and chlorhexidine (CHX) were used as positive controls. In addition, HOCl was examined for L929 cytotoxicity and RAW 264.7 growth.

The bacteriostatic ratio of NaOCl was comparable to that of CHX and significantly (P < 0.05) higher than that of all HOCl solutions. Higher HOCl concentration had significantly (P < 0.05) higher antibacterial effect, and the bacteriostatic ratio of 10 min treatment was slightly higher than that of 1 min treatment. CHX and NaOCl seeded into L929 cells resulted in low cell viability with only 30-39%, much significantly (P < 0.05) lower than all HOCl groups (greater than 80%). All HOCl solutions met the recommendations of ISO 10993-5 and showed no cytotoxicity, although there was a concentration-dependent decrease in cell viability. All antimicrobial agents showed the same trend of response to RAW 264.7 as L929.

Within the limit of this study, 400 ppm HOCl disinfectant may be a potential antimicrobial candidate for mouthwash, endodontic irrigants, and periodontitis treatment.