Nano-graphene oxide with antisense walR RNA inhibits the pathogenicity of Enterococcus faecalis in periapical periodontitis

BACKGROUND/PURPOSE: Enterococcus faecalis (E. faecalis) is considered a predominant pathogen for persistent periapical infections. Antisense walR (ASwalR) RNA was reported to inhibit the biofilm formation and sensitized E. faecalis to calcium hydroxide medication. The aims of this study were to investigate whether the graphene oxide (GO) nanosheets could be used to enhance antibacterial activity of ASwalR RNA for E. faecalis in periapical periodontitis. MATERIALS AND METHODS: We developed a graphene-based plasmid transformation system by loading antisense walR plasmid with GO-polyethylenimine (PEI) complexes (GO-PEI-ASwalR). The particle size distributions and zeta-potential of the GO-PEI-ASwalR were evaluated. Then, ASwalR plasmids were labeled with gene encoding enhanced green fluorescent protein (ASwalR-eGFP). The transformation efficiencies and the bacterial viability of E. faecalis were evaluated by confocal laser scanning microscopy. Quantitative real-time PCR assays were used to investigate the expressions of E. faecalis virulent genes after transformed by GO-PEI-ASwalR. Also, the antibacterial properties of the GO-PEI-ASwalR were validated in the rat periapical periodontitis model. RESULTS: We showed that GO-PEI could efficiently deliver the ASwalR plasmid into E. faecalis cell. GO-PEI-ASwalR significantly reduced virulent-associated gene expressions. Furthermore, GO-PEI-ASwalR suppressed biofilm aggregation and improved bactericidal effects using infected canal models in vitro. In four-weeks periapical infective rat models, the GO-PEI-ASwalR strains remarkably reduced the periapical lesion size. CONCLUSION: Transformation efficiency and antibacterial prosperity of ASwalR can be marked improved by GO-PEI based delivery system for E. faecalis infections.