Antibiofilm effect of ozonized physiological saline solution on peri-implant–related biofilm

Background: Removal of dental plaque and local application of local chemical adjuncts, such as chlorhexidine (CHX), have been used to control and treat peri-implant disease. However, these methods can damage the surface properties of the implants or promote bacterial resistance. The application of ozone as an adjunctive treatment represents a new approach in the management of peri-implantitis. Thus, the purpose of this study was to evaluate the antimicrobial effect of ozonized physiological saline solution in different concentrations against oral biofilms developed on titanium surface. Methods: Single and multi-species biofilms of Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus oralis were formed on titanium specimens for 5 days in anaerobic conditions. Biofilms were treated with ozonized saline solution at different concentrations (25, 50, and 80 μg/NmL), for 30 seconds and 1 minute. CHX (0.12%) and saline solution (0.89% NaCl) were used as positive and negative controls, respectively. Bacterial viability was quantified by colony forming units (CFU mL⁻¹), and biofilm images were acquired by confocal laser scanning microscopy (CLSM). Data were analyzed by parametric test (ANOVA) with Tukey post-hoc test (P < 0.05). Results: Ozonized saline solution showed antibiofilm activity at a concentration of 80 μg/NmL for 30 seconds and 1 minute, reducing, mainly, Porphyromonas gingivalis viability, with 2.78 and 1.7 log₁₀ CFU mL⁻¹ of reduction in both single and multi-species biofilms, respectively, when compared to the control (saline), whereas CHX reduced 1.4 and 1.2 log₁₀ CFU mL⁻¹. Conclusion: Ozonized saline solution has antibiofilm activity, with better effect when applied for 1 minute at 80 μg/NmL, being a promising candidate therapy for the treatment of peri-implant diseases.