Anti-biofilm and bystander effects of antimicrobial photo-sonodynamic therapy against polymicrobial periopathogenic biofilms formed on coated orthodontic mini-screws with zinc oxide nanoparticles.

ABSTRACT

BACKGROUND: The present study evaluated the anti-biofilm and bystander effects of antimicrobial photo-sonodynamic therapy (aPSDT) on the polymicrobial periopathogenic biofilms formed on mini-screws coated with zinc oxide nanoparticles (ZnONPs). MATERIALS AND METHODS: Thirty orthodontic identical mini-screws were divided into 6 groups (n = 5) as follows 1. negative control uncoated mini-screw + phosphate-buffered saline (PBS), 2. positive control uncoated mini-screw + 0.2% CHX, 3. coating control coated mini-screw + PBS, 4. antimicrobial photodynamic therapy (aPDT) coated mini-screw+light emitting diode (LED), 5. Antimicrobial sonodynamic therapy (aSDT) coated mini-screw+ultrasound waves, and 6. aPSDT coated mini-screw+LED+ultrasound waves. Electrostatic spray-assisted vapor deposition was employed to coat ZnONPs on titanium mini-screws. The biofilm inhibition test was used to assess the anti-biofilm efficacy against polymicrobial periopathogenic biofilms including Porphyromonas gingivitis, Prevotella intermedia, and Aggregatibacter actinomycetemcomitans, and the results were shown as the percent reduction of Log10 colony-forming unit (CFU)/mL. Following each treatment, the gene expression levels of TNF-α, IL-1ß, and IL-6 were evaluated on human gingival fibroblast (HGF) cells via quantitative real-time polymerase chain reaction (qRT-PCR) to reveal the bystander effects of aPSDT on HGF cells. RESULTS: A significant reduction in log10 CFU/mL of periopathogens was observed in groups treated with aPDT, aSDT, aPSDT, and 0.2% CHX up to 6.81, 6.63, 5.02, and 4.83 log, respectively, when compared with control groups (P<0.05). 0.2% CHX and aPSDT groups demonstrated significantly higher capacity in eliminating the periopathogen biofilm compared with other groups (P<0.05). The qRT-PCR showed that the expression level of inflammatory cytokines was significantly down regulated in aPDT, aSDT, and aPSDT groups (P<0.05). CONCLUSION: It was found that the ZnONPs-mediated aPSDT could significantly reduce periopathogen biofilm as well as the expression level of inflammatory cytokines.