Musculoskeletal| Volume 171, ISSUE 1, e61-e68, November 2011

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Evaluation and Comparison of the Effects of Hyperbaric Oxygen and Ozonized Oxygen as Adjuvant Treatments in an Experimental Osteomyelitis Model


      We evaluated and compared the efficacy of ozone (O3) and hyperbaric oxygen (HBO) therapies in an experimental rat model of osteomyelitis.

      Materials and Methods

      Forty-eight male Sprague-Dawley rats were divided into sham, osteomyelitis (control), vancomycin (V), vancomycin + HBO (VHB), vancomycin + O3 (VO), and vancomycin + HBO + O3 (VOHB) groups. Osteomyelitis was induced by a bone injection of 108 CFU/mL methicillin-resistant Staphylococcus aureus. HBO was administered daily at 2.8-atm pressure for 90 min; O3 therapy was provided as intraperitoneal injections of 0.7 mg/kg O3/O2 gas mixture once daily. Treatments were continued from d 7 to 21 after induction of osteomyelitis. Bone tissues and blood samples were harvested for biochemical, histopathologic, and microbiologic analyses.


      Rats in the sham, VO, and VOHB groups gained weight but those in the control, V, and VHB groups did not. Levels of malondialdehyde, superoxide dismutase, and glutathione peroxidase were lower in the VHB, VO, and VOHB groups than in V and control groups. Levels of interleukin-10 and -1β and tumor necrosis factor-α were decreased in the VHB, VO, and VOHB groups; transforming growth factor-β was increased in these groups compared with V and control groups (P ≤ 0.001). Bacteria counts in VOHB were significantly lower than those in group of V (P = 0.012). Histopathologic scores in group VO were significantly lower than those in group V (P = 0.046).


      O3 was as effective as HBO in decreasing oxidative parameters and inflammatory cytokines. Rats in the VO and VOHB groups gained more weight than did the other groups. Bacteria counts were significantly decreased in group VOHB compared with the other groups. Histopathologic scores in group VO were significantly decreased compared with the other groups.

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        • Tiemann A.H.
        • Hofmann G.O.
        Principles of the therapy of bone infections in adult extremities: Are there any new developments?.
        Strat Traum Limb Recon. 2009; 4: 57
        • Fritz J.M.
        • McDonald J.R.
        Osteomyelitis: Approach to diagnosis and treatment.
        Phys Sportsmed. 2008; 36: 116823
        • Stallmann H.P.
        • Faber C.
        • Bronckers A.L.
        • et al.
        Osteomyelitis prevention in rabbits using antimicrobial peptide hLF1-11- or gentamicin-containing calcium phosphate cement.
        J Antimicrob Chemother. 2004; 54 (Epub July 1, 2004): 472
        • Mader J.T.
        • Guckian J.C.
        • Glass D.L.
        • et al.
        Therapy with hyperbaric oxygen for experimental osteomyelitis due to Staphylococcus aureus in rabbits.
        J Infect Dis. 1978; 138: 312
        • Mader J.T.
        • Adams K.R.
        • Wallace W.R.
        • et al.
        Hyperbaric oxygen as adjunctive therapy for osteomyelitis.
        Infect Dis Clin North Am. 1990; 4: 433
        • Yıldız S.
        • Pehlivan O.
        • Ay H.
        • et al.
        Hyperbaric oxygen therapy in chronic osteomyelitis.
        Gulhane Med J. 2004; 46: 189
        • Mader J.T.
        • Brown G.L.
        • Guckian J.C.
        • et al.
        A mechanism for the amelioration by hyperbaric oxygen of experimental staphylococcal osteomyelitis in rabbits.
        J Infect Dis. 1980; 142: 915
        • Re L.
        • Mawsouf M.N.
        • Menéndez S.
        • et al.
        Ozone therapy: Clinical and basic evidence of its therapeutic potential.
        Arch Med Res. 2008; 39: 17
        • Bocci V.
        Ozone as Janus: This controversial gas can be either toxic or medically useful.
        Mediators Inflamm. 2004; 13: 3
        • Bocci V.
        Scientific and medical aspects of ozone therapy. State of the art.
        Arch Med Res. 2006; 37: 425
        • Oter S.
        • Korkmaz A.
        Relevance of hyperbaric oxygen to ozone therapy.
        Arch Med Res. 2006; 37: 917
        • Rice R.G.
        • Robson M.
        • Miller G.W.
        • et al.
        Uses of ozone in drinking water treatment.
        J Am Water Works Assoc. 1981; 73: 44
        • Steinhart H.
        • Schulz S.
        • Mutters R.
        Evaluation of ozonated oxygen in an experimental animal model of osteomyelitis as a further treatment option for skull-base osteomyelitis.
        Eur Arch Otorhinolaryngol. 1999; 256: 153
        • Dubovskiy I.M.
        • Martemyanov V.V.
        • Vorontsova Y.L.
        • et al.
        Effect of bacterial infection on antioxidant activity and lipid peroxidation in the midgut of Galleria mellonella L. larvae (Lepidoptera, Pyralidae).
        Comp Biochem Physiol C Toxicol Pharmacol. 2008; 148: 1
        • Paglia D.E.
        • Valentine W.N.
        Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase.
        J Lab Clin Med. 1967; 70: 158
        • García-Alvarez F.
        • Monzón M.
        • Grasa J.M.
        • et al.
        Interleukin-1, interleukin-6, and interleukin-10 responses after antibiotic treatment in experimental chronic Staphylococcus aureus osteomyelitis.
        J Orthop Sci. 2006; 11: 370
        • Galluzzo M.L.
        • Hernandez C.
        • Davila M.T.
        • et al.
        Clinical and histopathological features and a unique spectrum of organisms significantly associated with chronic granulomatous disease osteomyelitis during childhood.
        Clin Infect Dis. 2008; 46: 745
        • Aragón-Sánchez F.J.
        • Cabrera-Galván J.J.
        • Quintana-Marrero Y.
        • et al.
        Outcomes of surgical treatment of diabetic foot osteomyelitis: A series of 185 patients with histopathological confirmation of bone involvement.
        Diabetologia. 2008; 51: 1962
        • Darouiche R.O.
        • Landon G.C.
        • Patti J.M.
        • et al.
        Role of Staphylococcus aureus surface adhesins in orthopaedic device infections: Are results model-dependent?.
        J Med Microbiol. 1997; 46: 75
        • Kalteis T.
        • Beckmann J.
        • Schröder H.J.
        • et al.
        Moxifloxacin superior to vancomycin for treatment of bone infections–A study in rats.
        Acta Orthop. 2006; 77: 315
        • Monzon M.
        • Garcia-Alvarez F.
        • Lacleriga A.
        • et al.
        Evaluation of four experimental osteomyelitis infection models by using precolonized implants and bacterial suspensions.
        Acta Orthop Scand. 2002; 73: 11
        • Ohkawa H.
        • Ohishi N.
        • Yagi K.
        Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction.
        Anal Biochem. 1979; 95: 351
        • Durak I.
        • Yurtarslanl Z.
        • Canbolat O.
        • et al.
        A methodological approach to superoxide dismutase (SOD) activity assay based on inhibition of nitroblue tetrazolium (NBT) reduction.
        Clin Chim Acta. 1993; 214: 103
        • Kanellakopoulou K.
        • Thivaios G.C.
        • Kolia M.
        • et al.
        Local treatment of experimental Pseudomonas aeruginosa osteomyelitis with a biodegradable dilactide polymer releasing ciprofloxacin.
        Antimicrob Agents Chemother. 2008; 52: 2335
        • Mader J.T.
        • Adams K.
        Comparative evaluation of daptomycin (LY146032) and vancomycin in the treatment of experimental methicillin-resistant Staphylococcus aureus osteomyelitis in rabbits.
        Antimicrob Agents Chemother. 1989; 33: 689
        • Mader J.T.
        • Adams K.
        • Morrison L.
        Comparative evaluation of cefazolin and clindamycin in the treatment of experimental Staphylococcus aureus osteomyelitis in rabbits.
        Antimicrob Agents Chemother. 1989; 33: 1760
        • Hać S.
        • Dobosz M.
        • Kaczor J.J.
        • et al.
        Neutrophil engagement and septic challenge in acute experimental pancreatitis in rats.
        World J Gastroenterol. 2005; 11: 6459
        • Narayana K.
        An aminoglycoside antibiotic gentamycin induces oxidative stress, reduces antioxidant reserve and impairs spermatogenesis in rats.
        J Toxicol Sci. 2008; 33: 85
        • Fierbinţeanu-Braticevici C.
        • Mohora M.
        • Creţoiu D.
        • et al.
        Role of oxidative stress in the pathogenesis of chronic hepatitis C (CHC).
        Rom J Morphol Embryol. 2009; 50: 407
        • Saini A.
        • Sharma S.
        • Chhibber S.
        Protective efficacy of Emblica officinalis against Klebsiella pneumoniae induced pneumonia in mice.
        Indian J Med Res. 2008; 128: 188
        • Bocci V.
        Does ozone therapy normalize the cellular redox balance?.
        Med Hypot. 1996; 46: 150
        • Rodriguez Z.Z.
        • Guanche D.
        • Alvarez R.G.
        • et al.
        Preconditioning with ozone/oxygen mixture induces reversion of some indicators of oxidative stress and prevents organic damage in rats with fecal peritonitis.
        Inflamm Res. 2009; 58: 371
        • Guven A.
        • Gundogdu G.
        • Vurucu S.
        • et al.
        Medical ozone therapy reduces oxidative stress and intestinal damage in an experimental model of necrotizing enterocolitis in neonatal rats.
        J Pediatr Surg. 2009; 44: 1730
        • Madej P.
        • Plewka A.
        • Madej J.A.
        • et al.
        Ozonotherapy in an induced septic shock. I. Effect of ozonotherapy on rat organs in evaluation of free radical reactions and selected enzymatic systems.
        Inflammation. 2007; 30: 52
        • Schulz S.
        • Steinhart H.
        • Mutters R.
        • et al.
        Chronic osteomyelitis in a modified rabbit model.
        J Invest Surg. 1996; 9: 181
        • Yamayoshi T.
        • Tatsumi N.
        Microbicidal effects of ozone solution on methicillin-resistant Staphylococcus aureus.
        Drugs Exp Clin Res. 1993; 19: 59
      1. Sartori HE. Continously monitored high concentration ozonation for sterilization of blood and blood products. Canadian Patent CA2193418;1996.

        • Kekez M.M.
        • Sattar S.A.
        A new ozone-based method for virus inactivation: Preliminary study.
        Phys Med Biol. 1997; 42: 2027
      2. Ruan RR, Ma H, Chen PL, et al. Non-thermal disinfection of biological fluids using non-thermal plasma. United States Patent 20060127271 A1; 2006.

        • Burgassi S.
        • Zanardi I.
        • Travagli V.
        • et al.
        How much ozone bactericidal activity is compromised by plasma components?.
        J Appl Microbiol. 2009; 106: 1715
        • Torossian A.
        • Ruehlmann S.
        • Eberhart L.
        • et al.
        Pre-treatment with ozonized oxygen (O3) aggravates inflammation in septic rats.
        Inflamm Res. 2004; 53: S122
        • Bette M.
        • Nüsing R.M.
        • Mutters R.
        • et al.
        Efficiency of tazobactam/piperacillin in lethal peritonitis is enhanced after preconditioning of rats with O3/O2-pneumoperitoneum.
        Shock. 2006; 25: 23