In vitro studies of the antimicrobial effect of non-thermal plasma-activated water as a novel mouthwash

The aim of this study was to evaluate the antimicrobial effects of non-thermal plasma-activated water (PAW) as a novel mouthwash in vitro. Three representative oral pathogens - Streptococcus mutans, Actinomyces viscosus and Porphyromonas gingivalis - were treated with PAW. The inactivation effect was evaluated using the colony-forming unit (CFU) method, and the morphological and structural changes of a cell were observed by scanning electron microscopy and transmission electron microscopy (TEM). The physicochemical properties of PAW were analysed, and its influence on the leakage of intracellular proteins and DNA was evaluated. The results showed significant reduction of Streptococcus mutans within 60 s, of Actinomyces viscosus within 40 s, and of Porphyromonas gingivalis in less than 40 s. Scanning electron microscopy and TEM images showed that the normal cell morphology changed by varying degrees after treatment with PAW. Intracellular proteins (280 nm) and DNA (260 nm) leaked from all three species of bacteria after treatment with PAW. Reactive oxygen species (ROS), especially atomic oxygen (O), hydroxyl radical (OH), and hydrogen peroxide (H2O2), were generated and led to strong oxidative stress and cell damage. These results suggest that PAW has potential use as a novel antimicrobial mouthwash.