Advanced noninvasive light-activated disinfection: assessment of cytotoxicity on fibroblast versus antimicrobial activity against Enterococcus faecalis.

Recent interest in light-activated disinfection demands insight on the selectivity towards bacterial cells compared with mammalian cells. This study was aimed to evaluate the cytotoxicity and selectivity of an advanced noninvasive light-activated disinfection (ANILAD) developed in our laboratory. The extent of cytotoxic effect of methylene blue activated by visible light of wavelength 664 nm was tested and compared with sodium hypochlorite (NaOCl) under in vitro and ex vivo conditions on fibroblast L929 cells. Simultaneous evaluation of cytotoxicity and antibacterial effect was also conducted to study the specificity of light-activated therapy (LAT) toward prokaryotic cells (Enterococcus faecalis). The cytotoxicity was evaluated by 3-(4, 5-dimethylthiazol-2- yl)-2, 5-diphenyltetrazolium bromide assay and trypan blue viability test, whereas colony-forming units were determined to evaluate bacterial viability. Data from both in vitro and ex vivo experiments showed that cytotoxicity was significantly less in LAT compared with NaOCl (p<0.001). E faecalis cells were killed at a faster rate than fibroblasts. An irradiation dose producing 97.7% bacterial killing showed only 30% fibroblast dysfunction. This study indicated that ANILAD produced an insignificant effect on mammalian cells.