Abstract
Peri-implantitis is a bacteria-initiated infection that as yet has no effective treatment. A novel approach to treat peri-implantitis is the use of low-temperature plasma (LTP). LTP disrupts the biofilm while conditioning the surrounding host environment for bone growth around the infected implant. The goal of this study was to evaluate the antimicrobial properties of LTP on newly formed (24-h) and mature (7-days) peri-implant-related biofilms. Biofilm was composed of Actinomyces naeslundii (ATCC 12104), Porphyromonas gingivalis (W83), Streptococcus oralis (ATCC 35037), and Veillonella dispar (ATCC 17748). They were cultivated in brain heart infusion supplemented with 1% yeast extract, hemin (0.5 mg/mL), and menadione (5 mg/mL) and kept at 37⁰C in anaerobic conditions for 24-h. The species were mixed for a final concentration of ~105 colony forming units (CFU)/mL (OD=0.01), and the bacterial suspension was transferred to 24-well plates containing titanium specimens. Biofilms were treated with LTP for 1, 3, and 5 min at 3 or 10 mm from plasma-tip to sample. Controls were no treatment (Negative control=NC) and argon-flow at the same LTP conditions. Positive controls were 14 g/mL amoxicillin and 140 µg/mL metronidazole individually or combined, and 0.12% chlorhexidine. Biofilms were evaluated by CFU, confocal laser scanning microscopy (CLSM), and Fluorescence in situ Hybridization (FISH). Wilcoxon Signed-Rank and Wilcoxon Rank Sum tests were applied (α = 0.05). Bacterial growth was observed in all no-treatment groups corroborated by FISH. LTP treatment significantly reduced all bacteria species when compared to the NC in both tested periods and in all treatment combinations (p≤0.016), these results were corroborated by CLSM. There were no significant differences during biofilm development, between 24-h, 3, and 7 days within each LTP treatment, or among the bacteria within each LTP treatment (p≥0.05). LTP application is effective to reduce peri-implantitis-related multispecies biofilms on titanium surfaces.