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Effects of Red Deer Antlers on Cutaneous Wound Healing in Full-thickness Rat Models

  • Gu, LiJuan (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Mo, EunKyoung (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Yang, ZhiHong (Colloge of life science) ;
  • Fang, ZheMing (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Sun, BaiShen (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Wang, ChunYan (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Zhu, XueMei (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Bao, JianFeng (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University) ;
  • Sung, ChangKeun (Department of Food Science and Technology, College of Agriculture and Biotechnology Chungnam National University)
  • Received : 2007.06.02
  • Accepted : 2007.10.03
  • Published : 2008.02.01

Abstract

The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-${\beta}$(TGF-${\beta}$, insulin-like growth factor I (IGF- I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-${\beta}$ and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF- ${\beta}$ and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-${\beta}$ and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-${\beta}$(day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-${\beta}$ and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.

Keywords

Deer Antler;Full-thickness;Wound Healing;IGF-I;TGF-${\beta}$;EGF

References

  1. Akiko, K. K., K. M. Oda Yuko, A. M. Suzuki Masashi, O. Junko and I. Toru. 2005. Expression of fibroblast growth factors and their receptors during full-thickness skin wound healing in young and aged mice. J. Endocrinol. 186:273-289. https://doi.org/10.1677/joe.1.06055
  2. Benn, S. I., J. S. Whitsitt, K. N. Broadley, L. B. Nanney, D. Perkins, L. He, M. Patel, J. R. Morgan, W. F. Swain and J. M. Davidson. 1996. Particle-mediated gene transfer with transforming growth factor-beta1 cDNAs enhances wound repair in rat skin. J. Clin. Invest. 98:2894-2902. https://doi.org/10.1172/JCI119118
  3. Brown, G. L., L. Curtsinger, J. R. Brightwell, D. M. Ackerman, G. R. Tobin, H. C. Polk, Jr., C. George-Nascimento, P. Valenzuela and G. S. Schultz. 1986. Enhancement of epidermal regeneration by biosynthetic epidermal growth factor. J. Exp. Med. 163:1319-1324. https://doi.org/10.1084/jem.163.5.1319
  4. Buckley, A., J. M. Davidson, C. D. Kamerath, T. B. Wolt and S. C. Woodward. 1985. Sustained release of epidermal growth factor accelerates wound repair. Proc. Natl. Acad. Sci. USA. 82:7340-7344. https://doi.org/10.1073/pnas.82.21.7340
  5. Diegelmann, R. F. and M. C. Evans. 2004. Wound healing: an overview of acute, fibrotic and delayed healing. Front. Biosci. 9:283-289. https://doi.org/10.2741/1184
  6. Chen, C., J. Zhang, J. Li, J. Huang, C. Yang, G. Huang and J. Shi. 2004. Hydrodynamic-based in vivo transfection of retinoic X receptor-alpha gene can enhance vitamin A-induced attenuation of liver fibrosis in mice. Liver. Int. 24:679-686. https://doi.org/10.1111/j.1478-3231.2004.0977.x
  7. Elliott, J. L., J. M. Oldham, G. R. Ambler, J. J. Bass, G. S. Spencer, S. C. Hodgkinson, B. H. Breier, P. D. Gluckman and J. M. Suttie. 1992. Presence of insulin-like growth factor-I receptors and absence of growth hormone receptors in the antler tip. Endocrinol. 130:2513-2520. https://doi.org/10.1210/en.130.5.2513
  8. Feng, J. Q., D. Chen, J. Esparza, M. A. Harris, G. R. Mundy and S. E. Harris. 1995. Deer antler tissue contains two types of bone morphogenetic protein 4 mRNA transcripts. Biochim. Biophys. Acta. 1263:163-168. https://doi.org/10.1016/0167-4781(95)00106-Q
  9. Garcia, R. L., M. Sadighi, S. M. Francis, J. M. Suttie and J. S. Fleming. 1997. Expression of neurotrophin-3 in the growing velvet antler of the red deer Cervus elaphus. J. Mol. Endocrinol. 19:173-182. https://doi.org/10.1677/jme.0.0190173
  10. Ha, Y. W., B. T. Jeon, S. H. Moon, H. Toyoda, T. Toida, R. J. Linhardt and Y. S. Kim. 2005. Characterization of heparan sulfate from the unossified antler of Cervus elaphus. Carbohydr. Res. 340:411-416. https://doi.org/10.1016/j.carres.2004.11.011
  11. Gallant, C. L., M. E. Olson and D. A. Hart. 2004. Molecular, histologic, and gross phenotype of skin wound healing in red Duroc pigs reveals an abnormal healing phenotype of hypercontracted, hyperpigmented scarring. Wound. Repair. Regen. 12:305-319. https://doi.org/10.1111/j.1067-1927.2004.012311.x
  12. Franklin, J. D. and J. B. Lynch. 1979. Effects of topical applications of epidermal growth factor on wound healing. Experimental study on rabbit ears. Plast. Reconstr. Surg. 64:766-770. https://doi.org/10.1097/00006534-197912000-00003
  13. Francis, S. M. and J. M. Suttie. 1998. Detection of growth factors and proto-oncogene mRNA in the growing tip of red deer (Cervus elaphus) antler using reverse-transcriptase polymerase chain reaction (RT-PCR). J. Exp. Zool. 281:36-42. https://doi.org/10.1002/(SICI)1097-010X(19980501)281:1<36::AID-JEZ6>3.0.CO;2-D
  14. Hayashi, K., G. Frangieh, G. Wolf and K. R. Kenyon. 1989. Expression of transforming growth factor-beta in wound healing of vitamin A-deficient rat corneas. Invest. Ophthalmol. Vis. Sci. 30:239-247
  15. Haase, I., R. Evans, R. Pofahl and F. M. Watt. 2003. Regulation of keratinocyte shape, migration and wound epithelialization by IGF-1 and EGF dependent signalling pathways. J. Cell. Sci. 116:3227-3238. https://doi.org/10.1242/jcs.00610
  16. Jeon, B. T., M. H. Kim, S. M. Lee, David G. Thomas and S. H. Moon. 2006. Changes of Chemical Composition in Blood Serum during the Antler Growth Period in Spotted Deer (Cervus nippon). Asian-Aust. J. Anim. Sci. 19:1298-1304. https://doi.org/10.5713/ajas.2006.1298
  17. Kawano, M., S. Suzuki, M. Suzuki, J. Oki and T. Imamura. 2004. Bulge- and basal layer-specific expression of fibroblast growth factor-13 (FHF-2) in mouse skin. J. Invest. Dermatol. 122:1084-1090. https://doi.org/10.1111/j.0022-202X.2004.22514.x
  18. Jeon, B. T., S. K. Kang, S. M. Lee, S. K. Hong and S. H. Moon. 2007. Serum Biochemical Values during Antler Growth in Sika Deer (Cervus nippon). Asian-Aust. J. Anim. Sci. 20:748-753. https://doi.org/10.5713/ajas.2007.748
  19. Hiramatsu, M., K. Hatakeyama, N. Minami and M. Kumegawa. 1982. Increase in collagen synthesis of cotton pellet granuloma in rats by epidermal growth factor. Jpn. J. Pharmacol. 32:198-201. https://doi.org/10.1254/jjp.32.198
  20. Ignotz, R. A. and J. Massague. 1986 Transforming growth factorbeta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J. Biol. Chem. 261:4337-4345.
  21. Okumura, K., Y. Kiyohara, F. Komada, S. Iwakawa, M. Hirai and T. Fuwa. 1990. Improvement in wound healing by epidermal growth factor (EGF) ointment. I. Effect of nafamostat, gabexate, or gelatin on stabilization and efficacy of EGF. Pharm. Res. 7:1289-1293. https://doi.org/10.1023/A:1015946123697
  22. Newbrey, Jarrett W. and William J. Banks. 1983. Ultrastructural features of the cellular and matrical components of developing antler cartilage. In: Proceedings of the 1983 Kingsville Antler Development in Cervidae Symposium, Kingsville,Texas. pp. 261-272
  23. Montesano, R. and L. Orci. 1988. Transforming growth factor beta stimulates collagen-matrix contraction by fibroblasts: implications for wound healing. Proc. Natl. Acad. Sci. USA. 85:4894-4897 https://doi.org/10.1073/pnas.85.13.4894
  24. Lee, S. R., B. T. Jeon, S. J. Kim, M. H. Kim, S. M. Lee and S. H. Moon. 2007. Effects of Antler Development Stage on Fatty acid, Vitamin and GAGs Contents of Velvet Antler in Spotted Deer (Cervus nippon). Asian-Aust. J. Anim. Sci. 20:1546-1550. https://doi.org/10.5713/ajas.2007.1546
  25. Lee, E. H. and C. K. Joo. 1999. Role of transforming growth factor-$\beta$ in transdifferentiation and fibrosis of lens epithelial cells. Invest. Ophth. Vis. Sci. 40:2025-2032.
  26. Rappolee, D. A., D. Mark, M. J. Banda and Z. Werb. 1988. Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping. Sci. 24:708-712. https://doi.org/10.1126/science.24.622.708
  27. Robson, M. C., T. A. Mustoe and T. K. Hunt. 1998. The future of recombinant growth factors in wound healing. Am. J. Surg. 176 (2A Suppl):80S-82S
  28. Pierce, G. F., T. A. Mustoe, J. Lingelbach, V. R. Masakowski, P. Gramates and T. F. Deuel. 1989. Transforming growth factor beta reverses the glucocorticoid-induced wound-healing deficit in rats: possible regulation in macrophages by platelet-derived growth factor. Proc. Natl. Acad. Sci. USA. 86:2229-2233 https://doi.org/10.1073/pnas.86.7.2229
  29. Sporn, M. B., A. B. Roberts, J. H. Shull, J. M. Smith, J. M. Ward and J. Sodek. 1983. Polypeptide transforming growth factors isolated from bovine sources and used for wound healing in vivo. Sci. 219:1329-1331. https://doi.org/10.1126/science.6572416
  30. Suh, D. Y., T. K. Hunt and E. M. Spencer. 1992. Insulin-like growth factor-I reverses the impairment of wound healing induced by corticosteroids in rats. Endocrinol. 131:2399-2403. https://doi.org/10.1210/en.131.5.2399
  31. Singer, A. J. and Richard A. F. Clark. 1999. Cutaneous Wound Healing. Mech. Dis. 341:738-746.
  32. Toyokawa, H., Y. Matsui, J. Uhara, H. Tsuchiya, S. Teshima, H. Nakanishi, A. H. Kwon, Y. Azuma, T. Nagaoka, T. Ogawa and Y. Kamiyama. 2003. Promotive effects of far-infrared ray on full-thickness skin wound healing in rats. Exp. Biol. Med. (Maywood) 228:724-729. https://doi.org/10.1177/153537020322800612
  33. Tsunoda, S., H. Yamabe, H. Osawa, M. Kaizuka, K. Shirato and K. Okumura. 2001. Cultured rat glomerular epithelial cells show gene expression and production of transforming growth factorbeta: expression is enhanced by thrombin. Nephrol. Dial. Transplant. 16:1776-1782. https://doi.org/10.1093/ndt/16.9.1776
  34. Wang, Z. and J. Jiang. 2000. Wound healing research in the new century. Chin. Med. J. (Engl) 113:383-384.
  35. Bamman, M. M., J. R. Shipp, J. Jiang, B. A. Gower, G. R. Hunter, A. Goodman, C. L. McLafferty, Jr. and R. J. Urban. 2001. Mechanical load increases muscle IGF-I and androgen receptor mRNA concentrations in humans. Am. J. Physiol. Endocrinol. Metab. 280:E383-390. https://doi.org/10.1152/ajpendo.2001.280.3.E383
  36. Barling, P. M., A. K. Lai and L. F. Nicholson. 2005. Distribution of EGF and its receptor in growing red deer antler. Cell. Biol. Int. 29:229-236. https://doi.org/10.1016/j.cellbi.2004.12.004
  37. Kurtz, C. A., T. G. Loebig, D. D. Anderson, P. J. DeMeo and P. G. Campbell. 1999. Insulin-like growth factor I accelerates functional recovery from achilles tendon injury in a rat model. Am. J. Sport. Med. 27:363-369. https://doi.org/10.1177/03635465990270031701
  38. Theoret, C. L., S. M. Barber, T. N. Moyana and J. R. Gordon. 2001. Expression of transforming growth factor beta(1), beta(3), and basic fibroblast growth factor in full-thickness skin wounds of equine limbs and thorax. Vet. Surg. 30:269-277. https://doi.org/10.1053/jvet.2001.23341

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