Biocompatibility assessment of ozonated olive oil and common irrigating agents in regenerative endodontics: A cytotoxicity study on red blood cells

Introduction: The selection of appropriate treatment agents is critical in regenerative endodontics to ensure successful revascularization outcomes. Ozonated oil and water, known for their potent antibacterial and oxidation properties, offer a novel class of disinfectants. This study aims to evaluate the biocompatibility of ozonated olive oil and other commonly used irrigating solutions in regenerative endodontics, specifically focusing on their impact on red blood cells (RBCs). Objective: This research assesses the cytotoxic effects of ozonated olive oil and other irrigating solutions commonly employed in regenerative endodontics on red blood cells, aiming to identify potential alternatives with minimal adverse effects. Methods: Five test solution groups were established: Group I - 3% NaOCl, Group II - 1.5% NaOCl, Group III - 17% EDTA, Group IV - Ozonated olive oil, and Group V - Normal saline (control). Thirty-five test tubes containing diluted RBC suspensions were assigned randomly to these groups (seven per group). Each test tube received 100 microliters of the respective irrigant, followed by a 3-minute incubation and subsequent centrifugation. Hemoglobin (Hb%) concentration in the resulting supernatant was measured to estimate cytotoxicity. Statistical analyses employed Analysis of Variance and Tukey's Post Hoc analysis. Results: Among the test solutions, 3% NaOCl demonstrated the highest cytotoxicity, while normal saline exhibited the lowest, with mean Hb concentrations of 0.515 g/dl and 0.043 g/dl, respectively, due to hemolysis. Statistical analysis revealed significant differences among the various test groups (p = 0.001). Ozonated olive oil demonstrated cytotoxicity levels similar to saline, with no statistically significant difference between them (p = 0.988). Conclusion: In comparison to 3% NaOCl, 1.5% NaOCl, and 17% EDTA, ozonated olive oil displayed lower levels of cytotoxicity and released Hb concentration (0.086 g/dl). This suggests that ozonated olive oil holds promise as a potentially favorable adjunct irrigant for regenerative endodontic procedures, due to its reduced impact on red blood cells.