J Korean Med Sci.  2009 Jun;24(3):368-374. 10.3346/jkms.2009.24.3.368.

Therapeutic Effects of Topical Application of Ozone on Acute Cutaneous Wound Healing

Affiliations
  • 1Department of Dermatology, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea. yymmpark6301@hotmail.com

Abstract

This study was undertaken to evaluate the therapeutic effects of topical ozonated olive oil on acute cutaneous wound healing in a guinea pig model and also to elucidate its therapeutic mechanism. After creating full-thickness skin wounds on the backs of guinea pigs by using a 6 mm punch biopsy, we examined the wound healing effect of topically applied ozonated olive oil (ozone group), as compared to the pure olive oil (oil group) and non-treatment (control group). The ozone group of guinea pig had a significantly smaller wound size and a residual wound area than the oil group, on days 5 (P<0.05) and 7 (P<0.01 and P<0.05) after wound surgery, respectively. Both hematoxylin-eosin staining and Masson-trichrome staining revealed an increased intensity of collagen fibers and a greater number of fibroblasts in the ozone group than that in the oil group on day 7. Immunohistochemical staining demonstrated upregulation of platelet derived growth factor (PDGF), transforming growth factor-beta (TGF-beta) and vascular endothelial growth factor (VEGF) expressions, but not fibroblast growth factor expression in the ozone group on day 7, as compared with the oil group. In conclusion, these results demonstrate that topical application of ozonated olive oil can accelerate acute cutaneous wound repair in a guinea pig in association with the increased expression of PDGF, TGF-beta, and VEGF.

Keyword

Collagen; Ozone; Wound Healing

MeSH Terms

Acute Disease
Administration, Topical
Animals
Female
Guinea Pigs
Ozone/administration & dosage/*therapeutic use
Plant Oils/administration & dosage/therapeutic use
Platelet-Derived Growth Factor/metabolism
Skin/*drug effects/pathology
Transforming Growth Factor beta/metabolism
Vascular Endothelial Growth Factor A/metabolism
Wound Healing/*drug effects

Figure

  • Fig. 1 The effects of ozonated olive oil on clinical wound closure. The photomicrographs demonstrate the enhanced wound closure, in the ozone group, on the left two wounds (a and b) on the back of the guinea pig, as compared to the oil group (c) as well as the control group (d). (Bar=5 mm).

  • Fig. 2 Masson-trichrome staining of the wound bed and the edge of the injury site on days 3 and 7. The ozone group revealed the increased staining intensity of collagen fibers and the number of fibroblasts at the wound bed and edge, in comparison to the oil group and the control group on day 7, but not on day 3 (original magnification ×400, Bar=50 µm).

  • Fig. 3 Immunohistochemical staining for FGF, PDGF, TGF-β, and VEGF on day 7. The ozone group revealed the relatively increased expressions of PDGF (B, F, J), TGF-β (C, G, K), and VEGF (D, H, L), but not FGF (A, E, I) as compared to the oil group on day 7 (×100, original magnification, inlet; ×400, Bar=500 µm).


Reference

1. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev. 2003. 83:835–870.
Article
2. Ornitz DM, Itoh N. Fibroblast growth factors. Genome Biol. 2001. 2:REVIEWS3005.
3. Heldin CH, Westermark B. Mechanism of action and in vivo role of platelet-derived growth factor. Physiol Rev. 1999. 79:1283–1316.
Article
4. Lauer G, Sollberg S, Cole M, Flamme I, Sturzebecher J, Mann K, Krieg T, Eming SA. Expression and proteolysis of vascular endothelial growth factor is increased in chronic wounds. J Invest Dermatol. 2000. 115:12–18.
Article
5. Valacchi G, Fortino V, Bocci V. The dual action of ozone on the skin. Br J Dermatol. 2005. 153:1096–1100.
Article
6. Al-Dalain SM, Martinez G, Candelario-Jalil E, Menendez S, Re L, Giuliani A, Leon OS. Ozone treatment reduces markers of oxidative and endothelial damage in an experimental diabetes model in rats. Pharmacol Res. 2001. 44:391–396.
7. Martinez-Sanchez G, Al-Dalain SM, Menendez S, Re L, Giuliani A, Candelario-Jalil E, Alvarez H, Fernandez-Montequin JI, Leon OS. Therapeutic efficacy of ozone in patients with diabetic foot. Eur J Pharmacol. 2005. 523:151–161.
8. Bocci V. Ozone: a new medical drug. 2005. Dordrecht: Springer.
9. de Monte A, van der Zee H, Bocci V. Major ozonated autohemotherapy in chronic limb ischemia with ulcerations. J Altern Complement Med. 2005. 11:363–367.
10. Lim Y, Phung AD, Corbacho AM, Aung HH, Maioli E, Reznick AZ, Cross CE, Davis PA, Valacchi G. Modulation of cutaneous wound healing by ozone:differences between young and aged mice. Toxicol Lett. 2006. 160:127–134.
11. Gajendrareddy PK, Sen CK, Horan MP, Marucha PT. Hyperbaric oxygen therapy ameliorates stress-impaired dermal wound healing. Brain Behav Immun. 2005. 19:217–222.
Article
12. Janic B, Umstead TM, Phelps DS, Floros J. Modulatory effects of ozone on THP-1 cells in response to SP-A stimulation. Am J Physiol Lung Cell Mol Physiol. 2005. 288:L317–L325.
Article
13. Valacchi G, van der Vliet A, Schock BC, Okamoto T, Obermuller-Jevic U, Cross CE, Packer L. Ozone exposure activates oxidative stress responses in murine skin. Toxicology. 2002. 179:163–170.
Article
14. Bocci V. Biological and clinical effects of ozone. Has ozone therapy a future in medicine? Br J Biomed Sci. 1999. 56:270–279.
15. Valacchi G, Bocci V. Studies on the biological effects of ozone: 10. Release of factors from ozonated human platelets. Mediators Inflamm. 1999. 8:205–209.
Article
16. Lang DS, Jorres RA, Mucke M, Siegfried W, Magnussen H. Interactions between human bronchoepithelial cells and lung fibroblasts after ozone exposure in vitro. Toxicol Lett. 1998. 96-97:13–24.
Article
17. Sen CK, Khanna S, Babior BM, Hunt TK, Ellison EC, Roy S. Oxidant-induced vascular endothelial growth factor expression in human keratinocytes and cutaneous wound healing. J Biol Chem. 2002. 277:33284–33290.
Article
18. Sechi LA, Lezcano I, Nunez N, Espim M, Dupre I, Pinna A, Molicotti P, Fadda G, Zanetti S. Antibacterial activity of ozonized sunflower oil (Oleozon). J Appl Microbiol. 2001. 90:279–284.
Article
19. Fischer SM. Is cyclooxygenase-2 important in skin carcinogenesis? J Environ Pathol Toxicol Oncol. 2002. 21:183–191.
Article
20. Valacchi G, Pagnin E, Corbacho AM, Olano E, Davis PA, Packer L, Cross CE. In vivo ozone exposure induces antioxidant/stress-related responses in murine lung and skin. Free Radic Biol Med. 2004. 36:673–681.
Article
21. Paramio JM, Casanova ML, Segrelles C, Mittnacht S, Lane EB, Jorcano JL. Modulation of cell proliferation by cytokeratins K10 and K16. Mol Cell Biol. 1999. 19:3086–3094.
Article
22. Menendez S, Falcon L, Simon DR, Landa N. Efficacy of ozonized sunflower oil in the treatment of tinea pedis. Mycoses. 2002. 45:329–332.
23. Bocci V. Oxygen-ozone therapy: a critical evaluation. 2002. Dordrecht: Kluwer Academic Publisher.
24. FDA Wound Healing Clinical Focus Group. Guidance for industry: chronic cutaneous ulcer and burn wounds-developing products for treatment. Wound Repair Regen. 2001. 9:258–268.
25. Vincent F. Freedberg IM, Eisen AZ, Wolff K, Austen KF, Goldsmith LA, Katz SI, editors. Mechanisms of cutaneous wound repair. Fitzpatrick's dermatology in general medicine. 2003. 6th ed. New York: McGraw Hill;236–246.
26. Detmar M, Brown LF, Berse B, Jackman RW, Elicker BM, Dvorak HF, Claffey KP. Hypoxia regulates the expression of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) and its receptors in human skin. J Invest Dermatol. 1997. 108:263–268.
Article
27. Jazwa A, Loboda A, Golda S, Cisowski J, Szelag M, Zagorska A, Sroczynska P, Drukala J, Jozkowicz A, Dulak J. Effect of heme and heme oxygenase-1 on vascular endothelial growth factor synthesis and angiogenic potency of human keratinocytes. Free Radic Biol Med. 2006. 40:1250–1263.
Article
28. Nissen NN, Polverini PJ, Koch AE, Volin MV, Gamelli RL, DiPietro LA. Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing. Am J Pathol. 1998. 152:1445–1452.
29. Werner S, Peters KG, Longaker MT, Fuller-Pace F, Banda MJ, Williams LT. Large induction of keratinocyte growth factor expression in the dermis during wound healing. Proc Natl Acad Sci USA. 1992. 89:6896–6900.
Article
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