Effects of pure and ozonated sunflower seed oil (Helianthus annuus) on hypergranulation tissue formation, infection and healing of equine lower limb wounds

Main Article Content

Paula Alessandra Di Filippo
Luiza Maria Feitosa Ribeiro
Francielli Pereira Gobbi
Gabriela Bravim Lemos
Rachel Bittencourt Ribeiro
Hassan Jerdy
Larissa Carvalho da Silva
Inácio Silva Viana
Célia Raquel Quirino


This study was undertaken to evaluate the therapeutic effects of topical application of pure sunflower seed oil (oil group) and ozonated sunflower seed oil (ozone group) on acute cutaneous wound healing in eight healthy horses. The control group was treated with 0.9% sodium chloride. Two wounds were surgically produced on each horse on the proximal epiphysis of the metacarpus on the dorsal aspect of each forelimb. Wound area and contraction were measured on days 0, 3, 7, 14 and 21. Biopsy samples were taken to evaluate epithelial hyperplasia, inflammation, exuberant granulation tissue, exudate type, angiogenesis, and fibroplasia. Culture swabs were also collected. Wound contraction at 21 days for the oil group was 52.68%, for the ozone group was 72.16%, and for the control group (0.9% sodium chloride) was 34.80%. The ozone group had a significantly smaller wound size and a residual wound area than the control and the oil groups, on days 14 and 21. The control-wounds and oil-wounds had suppurative exudate and the presence of Streptococcus zooepidemicus. Neutrophilic inflammatory infiltrate was observed in all groups. Granulation tissue was observed on day 7 in all groups, but exuberant granulation tissue was observed only in the control group on days 14 and 21. Reepithelialization was observed on day 14 in the ozone group. In conclusion, these results demonstrate that topical application of ozonated sunflower seed oil accelerates acute cutaneous wound repair in horses, preventing hypergranulation tissue and infection, and that it is superior to treatment with pure sunflower seed oil and 0.9% sodium chloride.

Article Details

How to Cite
Di Filippo, P. A., Feitosa Ribeiro, L. M. ., Pereira Gobbi, F. ., Bravim Lemos, G. ., Bittencourt Ribeiro, R. ., Jerdy, H., Carvalho da Silva, L. ., Silva Viana, I. ., & Raquel Quirino, C. . (2020). Effects of pure and ozonated sunflower seed oil (Helianthus annuus) on hypergranulation tissue formation, infection and healing of equine lower limb wounds. Brazilian Journal of Veterinary Medicine, 42(1), e113520. https://doi.org/10.29374/2527-2179.bjvm1115321
Scientific articles


Al-Dalain, S. M., Martinez, G., Candelario-Jalil, E., Menendez, S., Re, L., Giuliani, A., & Leon, O. S. (2001). Ozone treatment reduces markers of oxidative and endothelial damage in an experimental diabetes model in rats. Pharmacological Research, 44(5), 391-396. http://dx.doi.org/10.1006/phrs.2001.0867.

Bertone, A. L. (1989). Management of exuberant granulation tissue. Veterinary Clinics of North America: Equine Practice, 5(3), 551-562. http://dx.doi.org/10.1016/s0749-0739(17)30574-6.

Burgassi, S., Zanardi, I., Travagli, V., Montomoli, E., & Bocci, V. (2009). How much ozone bactericidal activity is compromised by plasma components? Journal of Applied Microbiology, 106(5), 1715-1721. http://dx.doi.org/10.1111/j.1365-2672.2008.04141.x.

Caston, S. S. (2012). Wound care in horses. The Veterinary Clinics of North America. Equine Practice, 28(1), 83-100. http://dx.doi.org/10.1016/j.cveq.2012.01.001. PMid:22640581.

Crovetti, G., Martinelli, G., Issi, M., Barone, M., Guizzardi, M., Campanati, B., Moroni, M., & Carabelli, A. (2004). Platelet gel for healing cutaneous chronic wounds. Transfusion and Apheresis Science, 30(2), 145-151. http://dx.doi.org/10.1016/j.transci.2004.01.004. PMid:15062754.

Freeman, K., Woods, E., Welsby, S., Percival, S., & Cochrane, C. A. (2009). Biofilm evidence and the microbial diversity of horse wounds. Canadian Journal of Microbiology, 55(2), 197-202. http://dx.doi.org/10.1139/W08- 115. PMid:19295652.

Glasgow, W. C., & Eling, G. T. (1990). Epidermal growth factor simulates linoleic acid metabolic in BAB/C 3T3 fibroblasts. Molecular Pharmacology, 38(4), 503-510. PMID: 2233691.

Harman, R. M., Theoret, C. L., & Van de Walle, G. R. (2019). The horse as a model for the study of cutaneous wound healing. Advances in Wound Care, 1-19. In press. http://dx.doi.org/10.1089/wound.2018.0883.

Kim, H. S., Noh, S. U., Han, Y. W., Kim, K. M., Kang, H., Kim, H. O., & Park, Y. M. (2009). Therapeutic effects of topical application of ozone on acute cutaneous wound healing. Journal of Korean Medical Science, 24, 368-374. http://dx.doi.org/10.3346/jkms.2009.24.3.368.

Macri, L., & Clark, R. A. (2009). Tissue engineering for cutaneous wounds: Selecting the proper time and space for growth factors, cells and the extracellular matrix. Skin Pharmacology and Physiology, 22, 83-93. http://dx.doi.org/10.1159/000178867.

Magalhães, M. S., Fechine, F. V., Macedo, R. N., Monteiro, D. L., Oliveira, C. C., Brito, G. A., Moraes, M. E., & Moraes, M. O. (2008). Effect of a combination of medium chain triglycerides, linoleic acid, soy lecithin and vitamins A and E on wound healing in rats. Acta Cirurgica Brasileira, 23(3), 262-269. http://dx.doi.org/10.1590/S0102-86502008000300009. PMid:18552998.

Marques, S. R., Peixoto, C. A., Messias, J. B., Albuquerque, A. R., & Silva Junior, V. A. (2004). The effects of topical application of sunflower seed oil on open wound healing in lambs. Acta Cirurgica Brasileira, 19(3), 196-205. http://dx.doi.org/10.1590/S0102-86502004000300005.

Martinez-Ferrer, M., Afshar-Sherif, A. R., Uwamariya, C., Crombrugghe, B., Davidson, J. M., & Bhowmick, N. A. (2010). Dermal transforming growth factor-beta responsiveness mediates wound contraction and epithelial closure. The American Journal of Pathology, 176, 98-107. http://dx.doi.org/10.2353/ajpath.2010.090283.

Maxson, S., Lopez, E. A., Yoo, D., Danilkovitch-Miagkova, A., & Leroux, M. A. (2012). Concise review: Role of mesenchymal stem cells in wound repair. Stem Cells Translational Medicine, 1, 142-149. http://dx.doi.org/10.5966/sctm.2011-0018.

Morgan, D. D., McClure, S., Yaeger, M. J., Schumacher, J., & Evans, R. B. (2009). Effects of extracorporeal shock wave therapy on wounds of the distal portion of the limbs in horses. Journal of the American Veterinary Medical Association, 234(9), 1154-1161. http://dx.doi.org/10.2460/javma.234.9.1154. PMid:19405886.

Oliveira Junior, L. A. T., Souza, V. R. S., Endringer, D. C., Hendrickson, D. A., & Coelho, C. S. (2012). Effects of topical application of sunflower seed oil on experimentally induced wounds in horses. Journal of Equine Veterinary Science, 32(3), 139-145. http://dx.doi.org/10.1016/j.jevs.2011.08.006.

Ramsey, D. T., Pope, E. R., Wagner-Mann, C., Berg, J. N., & Swaim, S. F. (1995). Effects of three occlusive dressing materials on healing of full-thickness skin wounds in dogs. American Journal of Veterinary Research, 56(7), 941-949. PMid:7574165.

Theoret, C. L., Barber, S. M., Moyana, T. N., & Gordon, J. R. (2002). Preliminary observations on expression of transforming growth factors beta1 and beta3 in equine full-thickness skin wounds healing normally or with exuberant granulation tissue. Veterinary Surgery, 31, 266-273. http://dx.doi.org/10.1053/jvet.2002.32394.

Tracey, A. K., Alcott, C. J., Schleining, J. A., Safayi, S., Zaback, P. C., Hostetter, J. M., & Reinertson, E. L. (2014). The effects of topical oxygen therapy on equine distal limb dermal wound healing. The Canadian Veterinary Journal. La Revue Veterinaire Canadienne, 55(12), 1146-1152. PMid:25477541.

Travagli, V., Zanardi, I., Valacchi, G., & Bocci, V. (2010). Ozone and ozonated oils in skin diseases: a review. Mediators of Inflammation, 2010, 610418. http://dx.doi.org/10.1155/2010/610418. PMid:20671923.

Valacchi, G., Fortino, V., & Bocci, V. (2005). The dual action of ozone on the skin. British Journal of Dermatology, 153, 1096-100. http://dx.doi.org/10.1111/j.1365-2133.2005.06939.x

Valacchi, G., Lim, Y., Belmonte, G., Miracco, C., Zanardi, I., Bocci, V., & Travagli, V. (2011). Ozonated sesame oil enhances cutaneous wound healing in SKH1 mice. Wound Repair and Regeneration, 19, 107-115. http://dx.doi.org/10.1111/j.1524-475X.2010.00649.x.

Valacchi, G., Pagnin, E., Corbacho, A. M., Olano, E., Davis, P. A., Packer, L., & Cross, C. E. (2004). In vivo ozone exposure induces antioxidant/stress-related responses in murine lung and skin. Free Radical Biology and Medicine, 36, 673-681. http://dx.doi.org/10.1016/j.freeradbiomed.2003.12.005.

Westgate, S. J., Percival, S. L., Knottenbelt, D. C., Clegg, P. D., & Cochrane, C. A. (2011). Microbiology of equine wounds and evidence of bacterial biofilms. Veterinary Microbiology, 150(1-2), 152-159. http://dx.doi.org/10.1016/j. vetmic.2011.01.003. PMid:21273008.

Witzel, I. I., Koh, L. F., & Perkins, N. D. (2010). Regulation of cyclin D1 gene expression. Biochemical Society Transactions, 38(Pt 1), 217-222. http://dx.doi.org/10.1042/BST0380217. PMid:20074063.

Ziboh, V. A. (1996). The significance of polyunsaturated fatty acids in cutaneous biology. Lipids, 31(1, Suppl), S249-S53. http://dx.doi.org/10.1007/BF02637085. PMid:8729128.

Most read articles by the same author(s)