Objective: The aim of the present study was to investigate the biochemical and histopathological impact of ozone treatment in an experimental model of osteomyelitis in rats. Methods: A total of 24 adult male Sprague-Dawley rats (3 months old, each weighing 300 to 400 g) were randomly allocated into three groups. Group I (n=8) served as a control and received no interventions or medications. In Group II (n=8), osteomyelitis was induced in the femur and no treatment was applied. Group III (n=8) received intraperitoneal ozone treatment for 3 weeks after the formation of osteomyelitis in the femur. Serum samples were taken to assess total antioxidant capacity (TAC), protein carbonyl content (PCO), and lactate dehydrogenase (LDH). Bone specimens obtained from the femur were histopathologically evaluated for inflammation, necrosis, osteomyelitis, and abscess formation. Results: Serum TAC levels were notably higher (p<0.001), while LDH levels were lower (p=0.002) in Group III than Group II. No significant difference was detected between groups with respect to PCO level. Similarly, Group III displayed more favorable histopathological outcomes with respect to osteomyelitis (p=0.008), inflammation (p=0.001), necrosis (p=0.022), and abscess formation (p=0.022). Conclusion: Ozone may be a useful adjunct treatment for osteomyelitis. Further studies in animals and humans are needed to clarify and confirm these preventive effects, understand the underlying pathophysiology, and establish guidelines.