냉동보관 후 이식한 지방조직의 생물학적 변화와 생존에 관한 연구

A Study for the Biologic Changes and Viability of Adipose tissue in Cryopreserved Fat Graft

  • 한재정 (동아대학교 의과대학 성형외과학교실) ;
  • 권용석 (동아대학교 의과대학 성형외과학교실) ;
  • 이장호 (동아대학교 의과대학 성형외과학교실) ;
  • 허정 (동아대학교 의과대학 성형외과학교실) ;
  • 이근철 (동아대학교 의과대학 성형외과학교실) ;
  • 김명훈 (동아대학교 의과대학 성형외과학교실) ;
  • 김석권 (동아대학교 의과대학 성형외과학교실)
  • Han, Jae-Jeong (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Kwon, Yong-Seok (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Lee, Jang-Ho (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Heo, Jung (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Lee, Keun-Cheol (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Kim, Myung-Hoon (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University) ;
  • Kim, Seok-Kwun (Department of Plastic & Reconstructive Surgery, College of Medicine. Dong-A University)
  • 발행 : 2009.03.15

초록

Purpose: Adipose tissue injection as a free graft for the correction of soft - tissue deficiency or depression deformity is a widespread procedure in plastic surgery. This study is to analyze the changes and viability of cryopreserved adipose tissue and to find out efficient long - term storage period. Methods: After centrifugation of aspirated abdominal tissues, $10m{\ell}$ of packed Adipose tissue were freezed at $-20^{\circ}C$. For 2, 4, 6, 8 months, each frozen samples were taken and injected into scalp of SCID mice. After 15 weeks, injected Adipose tissue were sampled and analyzed at 2 months interval. We compared and analyzed each group about the weight of the injected fat, histologic impressions, activity of mitochondria, size of a fat cell and rate of survival. Results: Significant weight changes were observed in cryopreservation for 2 months(p<0.05). Histologic changes were observed, independent of the freezing period with H - E stain. Among cryopreservations for 2, 4, 6 months, no significant change were observed. The reduction of mitochondrial enzymatic activity was observed independent of time interval but activity of mitochondrial dehydrogenase was reduced less than 50% in MTT assay. Conclusion: Freezing in $-20^{\circ}C$ for 6 months has no adverse effect to Adipose tissue, but fragile adipocytes, damaged cell membrane during harvesting procedure, were disrupted within 1 - 2 month and the maximum volume reduction were followed less than 2 months. These results demonstrate that tissue preparation cells without membrane damage have the greatest viability level and cryopreservation less than 2 months has great volume effect and cryopreservation for 6 months has stable volume effect.

키워드

참고문헌

  1. Sommer B, Sattler G: Current concepts of fat graft survival: histology of aspirated adipose tissue and review of the literature. Dermatol Surg 26: 1159, 2000 https://doi.org/10.1046/j.1524-4725.2000.00278.x
  2. Rohrich RJ, Sorokin ES, Brown SA: In search of improved fat transfer viability: a quantitative analysis of the role of centrifugation and harvest site. Plast Reconstr Surg 113: 391, 2004 https://doi.org/10.1097/01.PRS.0000097293.56504.00
  3. Boschert MT, Beckert BW, Puckett CL, Concannon MJ: Analysis of lipocyte viability after liposuction. Plast Reconstr Surg 109: 761, 2002 https://doi.org/10.1097/00006534-200202000-00054
  4. MacRae JW, Tholpady SS, Ogle RC, Morgan RF: Ex vivo fat graft preservation: effects and implications of cryopreservation. Ann Plast Surg 52: 281, 2004 https://doi.org/10.1097/01.sap.0000110559.58945.2e
  5. MacRae MW, Tholpady SS, Katz AJ, GAmpper TG, Drake DB, Ogle RC, Morgan RF: Human adipocyte viability testing: a new assay. Aesthetic Surg J 23: 265, 2003 https://doi.org/10.1016/S1090-820X(03)00151-1
  6. Miller RR Jr, Sheffer CJ, Cornett CL, McClean R, MacCallum C, Johnston SD: Sperm membrane fatty acid composition in the eastern grey kangaroo(Macropus giganteus), koala(Phascolarctos cinereus) and common wombat(Vombatus ursinus) and its relationship to cold shock injury and cryopreservation success. Cryobiology 49: 137, 2004 https://doi.org/10.1016/j.cryobiol.2004.06.002
  7. Rendal Vazquez ME, Roman TD, Cuesta MG, Botta CZ, Ibanez JS, Diaz SP, Nunez CA: Viability and histologic structure of porcine valves after cryopreservation. Ann Thorac Surg 77: 186, 2004 https://doi.org/10.1016/S0003-4975(03)01491-7
  8. Lalikos JF, Li YQ, Roth TP, Doyle JW, Matory WE, Lawrence WT: Biochemical assessment of cellular damage after adipocyte harvest. J Surg Res 70: 95, 1997 https://doi.org/10.1006/jsre.1997.5090
  9. Muller G, Jordan H, Jung C, Kleine H, Perty S: Analysis of lipocytes in adipocytes using a fluorescent fatty acid derivative. Biochimie 85: 1245, 2003 https://doi.org/10.1016/j.biochi.2003.10.004
  10. Lu JH, Chiu YT, Sung HW, Hwang B, Chong CK, Chen SP, Mao SJ, Yang PZ, Chang Y: XTT-colorimetric assay as a marker of viability in cryoprocessed cardiac valve. J Mol Cell Cardiol 29: 1189, 1997 https://doi.org/10.1006/jmcc.1996.0354
  11. O'Connell M, McClure N, Lewis SE: The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 17: 704, 2002 https://doi.org/10.1093/humrep/17.3.704
  12. Kim YK, Park HS, Lee HJ: Studies on the proper storage period and change of -20°Ccryopreserved adipocyte. J Korean Soc Aesthtic Plast Surg 12: 33, 2006