Effect of Relaxin Expressing Adenovirus for Rat Skin Flap Viability

Relaxin을 분비하는 아데노바이러스가 피판의 생존에 미치는 영향

  • Yun, In-Sik (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine) ;
  • Park, Yong-Sun (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine) ;
  • Cheon, Young-Woo (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine) ;
  • Jeon, Yeo-Reum (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine) ;
  • Lee, Won-Jai (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine) ;
  • Yun, Chae-Ok (Yonsei Cancer Research Center, Yonsei University College of Medicine) ;
  • Rah, Dong-Kyun (Institue for Human Tissue Restoration & Department of Plastic & Reconstructive Surgery Yonsei University College of Medicine)
  • 윤인식 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 박용순 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 전영우 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 전여름 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 이원재 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 윤채옥 (연세대학교 의과대학 성형외과학교실 연세암연구소) ;
  • 나동균 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소)
  • Received : 2010.05.07
  • Accepted : 2010.07.19
  • Published : 2010.09.10

Abstract

Purpose: Of various effects of relaxin, we assumed that anti-fibrotic effects, neovascularization effects and vasodilatation effects of relaxin might enhance the survival rate of skin flap. In the current study, we used adenovirus expressing relaxin genes to examine whether these genes could enhance the survival rate of a skin flap. Methods: A total of 30 Sprangue-Dawley rats were divided into three groups: RLX group (10; relaxin virus injected group), CTR group (10; no gene coded virus injection group), and PBS group (10; PBS injected group). Each group was intradermally injected with the virus ($10^7$ PFU) and PBS 48 hours before and immediately before the flap elevation. A distally based flap $3{\times}9\;cm$ in size was elevated on the dorsal aspect of each rat. Following this, a flap was placed in the original location and then sutured using a #4-0 Nylon. A surviving area of the flap was measured and then compared on postoperative days 3, 7 and 10. Using a laser Doppler, the amount of blood flow was measured. On postoperative day 10, tissues were harvested for histologic examination and the number of blood vessels was counted. Results: There was a significant increase in the area of the flap survival in the RLX group on postoperative days 3 and 7. The Doppler measurement also showed significantly increased blood flow immediately after the operation and on postoperative days 7 and 10. The number of blood vessels was significantly greater in the RLX group in the tissue harvested on postoperative day 10. The VEGF concentration was significantly higher in the RLX group than others in the tissues harvested on postoperative day 10. Conclusion: Following an analysis of the effects of relaxin-secreting adenovirus on the survival of a flap, the surviving area of the flap and the blood flow also increased. A histopathology also showed an increase in the number of blood vessels and the concentration of VEGF.

Keywords

References

  1. Sherwood OD: Relaxin's physiological roles and other diverse actions. Endocr Rev 25: 205, 2004 https://doi.org/10.1210/er.2003-0013
  2. Unemori EN, Erikson ME, Rocco SE, Sutherland KM, Parsell DA, Mak J, Grove BH: Relaxin stimulates expression of vascular endothelial growth factor in normal human endometrial cells in vitro and is associated with meno-metrorrhagia in women. Hum Reprod 14: 800, 1999 https://doi.org/10.1093/humrep/14.3.800
  3. Samuel CS, Hewitson TD, Unemori EN, Tang ML: Drugs of the future: the hormone relaxin. Cell Mol Life Sci 64: 1539, 2007 https://doi.org/10.1007/s00018-007-6543-y
  4. Kim JH, Lee YS, Kim H, Huang JH, Yoon AR, Yun CO: Relaxin expression from tumor-targeting adenoviruses and its intratumoral spread, apoptosis induction, and efficacy. J Natl Cancer Inst 98: 1482, 2006 https://doi.org/10.1093/jnci/djj397
  5. Khan A, Ashrafpour H, Huang N, Neligan PC, Kontos C, Zhong A, Forrest CR, Pang CY: Acute local subcutaneous VEGF165 injection for augmentation of skin flap viability: efficacy and mechanism. Am J Physiol Regul Integr Comp Physiol 287: R1219, 2004 https://doi.org/10.1152/ajpregu.00143.2004
  6. Kerrigan CL: Skin flap failure: pathophysiology. Plast Reconstr Surg 72: 766, 1983 https://doi.org/10.1097/00006534-198312000-00003
  7. Hopper RA, Forrest CR, Xu H, Zhong A, He W, Rutka J, Neligan P, Pang CY: Role and mechanism of PKC in ischemic preconditioning of pig skeletal muscle against infarction. Am J Physiol Regul Integr Comp Physiol 279: R666, 2000 https://doi.org/10.1152/ajpregu.2000.279.2.R666
  8. Maldonado C, Stadelmann WK, Ramirez S, Quan EE, Barker JH: Preconditioning of latissimus dorsi muscle flaps with monophosphoryl lipid a. Plast Reconstr Surg 111: 267, 2003 https://doi.org/10.1097/00006534-200301000-00045
  9. Seify H, Bilkay U, Jones G: Improvement of TRAM flap viability using human VEGF-induced angiogenesis: a comparative study of delay techniques. Plast Reconstr Surg 112: 1032, 2003 https://doi.org/10.1097/01.PRS.0000076186.97093.92
  10. Scalise A, Tucci MG, Lucarini G, Giantomassi F, Orlando F, Pierangeli M, Pugnaloni A, Bertani A, Ricotti G, Biagini G: Local rh-VEGF administration enhances skin flap survival more than other types of rh-VEGF administration: a clinical, morphological and immunohistochemical study. Exp Dermatol 13: 682, 2004 https://doi.org/10.1111/j.0906-6705.2004.00220.x
  11. Shima DT, Deutsch U, D'Amore PA: Hypoxic induction of vascular endothelial growth factor (VEGF) in human epithelial cells is mediated by increases in mRNA stability. FEBS Lett 370: 203, 1995 https://doi.org/10.1016/0014-5793(95)00831-S
  12. Huang N, Khan A, Ashrafpour H, Neligan PC, Forrest CR, Kontos CD, Pang CY: Efficacy and mechanism of adeno-virus-mediated VEGF-165 gene therapy for augmentation of skin flap viability. Am J Physiol Heart Circ Physiol 291: H127, 2006 https://doi.org/10.1152/ajpheart.01253.2005
  13. Bani D, Maurizi M, Bigazzi M: Original article: relaxin reduces the number of circulating platelets and depresses platelet release from megakaryocytes: studies in rats. Platelets 6: 330, 1995 https://doi.org/10.3109/09537109509078467
  14. Bani D, Bigazzi M, Masini E, Bani G, Sacchi TB: Relaxin depresses platelet aggregation: in vitro studies on isolated human and rabbit platelets. Lab Invest 73: 709, 1995
  15. Zheng Y, Yi C, Xia W, Ding T, Zhou Z, Han Y, Guo S: Mesenchymal stem cells transduced by vascular endothelial growth factor gene for ischemic random skin flaps. Plast Reconstr Surg 121: 59, 2008 https://doi.org/10.1097/01.prs.0000293877.84531.5a