Comparisons of the killing effect of direct current partially mediated by reactive oxygen species on Porphyromonas gingivalis and Prevotella intermedia in planktonic state and biofilm state – an in vitro study

Abstract

Background/purpose: Bacterial biofilms formed on the surface of tissues and biomaterials are major causes of chronic infections in humans. Among them, Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia) are anaerobic pathogens causing dental infections associated with periodontitis. In this study, we evaluated the killing effect and underlying mechanisms of direct current (DC) as an antimicrobial method in vitro. Materials and methods: We chose P. gingivalis and P. intermedia in different states to make comparisons of the killing effect of DC. By viable bacteria counting, fluorescent live/dead staining, reactive oxygen species (ROS) assay, addition of ROS scavenger DMTU and mRNA expression assay of ROS scavenging genes, the role of ROS in the killing effect was explored. Results: The planktonic and biofilm states of two bacteria could be effectively killed by low-intensity DC. For the killing effect of 1000 μA DC, there were significant differences whether on planktonic P. gingivalis and P. intermedia (mean killing values: 2.40 vs 2.62 log10 CFU/mL) or on biofilm state of those (mean killing values: 0.63 vs 0.98 log10 CFU/mL). 1000 μA DC greatly induced ROS production and the mRNA expression of ROS scavenging genes. DMTU could partially decrease the killing values of DC and downregulate corresponding gene’s expression. Conclusion: 1000 μA DC can kill P. gingivalis and P. intermedia in two states by promoting overproduction of ROS, and P. intermedia is more sensitive to DC than P. gingivalis. These findings indicate low-intensity DC may be a promising approach in treating periodontal infections.

Keywords

• Direct current
• Porphyromonas gingivalis
• Prevotella intermedia
• Biofilm
• Reactive oxygen species
• Killing effect