Archives of Plastic Surgery

Korean Society of Plastic and Reconstructive Surgeons (대한성형외과학회)
권호 표지

Optimal Concentration of Thrombin to Activate Platelet for Wound Healing

창상치유 목적의 혈소판 치료를 위한 Thrombin의 최적 농도

  • Eum, Soo Jin (Department of Plastic Surgery Korea University College of Medicine) ;
  • Han, Seung Kyu (Department of Plastic Surgery Korea University College of Medicine) ;
  • Chun, Kyung Wook (Department of Plastic Surgery Korea University College of Medicine) ;
  • Kim, Woo Kyung (Department of Plastic Surgery Korea University College of Medicine)
  • 엄수진 (고려대학교 의과대학 성형외과학교실) ;
  • 한승규 (고려대학교 의과대학 성형외과학교실) ;
  • 전경욱 (고려대학교 의과대학 성형외과학교실) ;
  • 김우경 (고려대학교 의과대학 성형외과학교실)
  • Published : 2009.01.15
⟨ Previous Next ⟩

Abstract

Purpose: Platelet transplantation is a novel therapeutic strategy for acceleration of wound healing. When applying platelets, efficacy of adding thrombin to stimulate growth factor release from platelets has already been proved. However, no quantitative data of the thrombin treatment has been reported. The purpose of this study was to determine the optimal thrombin concentration to maximize growth factor release of platelets. In particular, this study was designed to quantify levels of platelet derived growth factor(PDGF)-BB, which is a major growth factor contained in the platelets, in vitro. Methods: Fresh platelets were obtained from a blood bank. They were suspended in DMEM/F - 12 and incubated with thrombin of various concentrations. The concentrations of thrombin tested were 0, 6.25, 12.5, 25, 50, 100, 200, and 400 IU/ml. After 30 minutes, 1, 3, 5, and 7 days, the levels of PDGF - BB were measured using enzyme linked immunosorbent assay. Platelets from four donors were included in this study. Each sample was tested in triplicate and the mean value was used as a data for each sample. Results: The addition of thrombin increased the level of PDGF - BB. Increases in storage time of platelets resulted in decreased levels of PDGF - BB. Higher levels of PDGF were detected in consort with increased thrombin concentrations. However, there was no significant difference between samples of 200 and 400 IU/ml concentrations. Conclusion: The results indicate that adding thrombin accelerates the release of growth factors from platelets and the optimal thrombin concentration to maximize this function is 200 IU/ml.

Keywords

References

  1. Han SK, Choi KJ, Kim WK: Clinical application of fresh fibroblast allografts for the treatment of diabetic foot ulcers: a pilot study. Plast Reconstr Surg 114: 1783, 2004 https://doi.org/10.1097/01.PRS.0000142415.57470.DF
  2. Crovetti G, Martinelli G, Issi M, Barone M, Guizzardi M, Campanati B, Moroni M, Carabelli A: Platelet gel for healing cutaneous chronic wounds. Transfus Apher Sci 30: 145, 2004 https://doi.org/10.1016/j.transci.2004.01.004
  3. Marx RE: Platelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg 62: 489, 2004 https://doi.org/10.1016/j.joms.2003.12.003
  4. Knighton DR, Ciresi KF, Fiegel VD, Austin LL, Butler EL: Classification and treatment of chronic nonhealing wounds. Successful treatment with autologous platelet- derived wound healing factors(PDWHF). Ann Surg 204: 322, 1986 https://doi.org/10.1097/00000658-198609000-00011
  5. Knighton DR, Ciresi K, Fiegel VD, Schumerth S, Butler E, Cerra F: Stimulation of repair in chronic, nonhealing, cutaneous ulcers using platelet-derived wound healing formula. Surg Gynecol Obstet 170: 56, 1990
  6. Krupski WC, Reilly LM, Perez S, Moss KM, Crombleholme PA, Rapp JH: A prospective randomized trial of autologous platelet-derived wound healing factors for treatment of chronic nonhealing wounds: a preliminary report. J Vasc Surg 14: 526, 1991 https://doi.org/10.1067/mva.1991.31288
  7. Margolis DJ, Kantor J, Santanna J, Strom BL, Berlin JA: Effectiveness of platelet releasate for treatment of diabetic neuropathic foot ulcer. Diabetes Care 24: 483, 2001 https://doi.org/10.2337/diacare.24.3.483
  8. Han SK, Kim DW, Jeong SH, Hong YT, Woo HS, Kim WK, Gottrup F: Potential use of blood bank platelet concentrates to accelerate wound healing of diabetic ulcer. Ann Plast Surg 59: 532, 2007 https://doi.org/10.1097/01.sap.0000258983.25728.0b
  9. Hong YT, Han SK, Lee BI, Kim WK: Level of Platelet Derived Growth Factor(PDGF) in Blood Bank Platelet Concentrate. J Korean Soc Plast Reconstr Surg 33: 732, 2006
  10. Greenhalgh DG: Wound healing and diabetes mellitus, Clin Plast Surg 30: 37, 2003 https://doi.org/10.1016/S0094-1298(02)00066-4
  11. Eppley BL, Woodell JE, Higgins J: Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plast Reconstr Surg 114: 1502, 2004
  12. Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR: Platelet-rich plasma: Growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 85: 638, 1998 https://doi.org/10.1016/S1079-2104(98)90029-4
  13. Puri RN: Phospholipase A2: its role in ADP-and thrombin-induced platelet activation mechanisms. Int J Biochem Cell Biol 30: 1107, 1998 https://doi.org/10.1016/S1357-2725(98)00080-6
  14. I. Martineau I, Lacoste E, Gagnon G: Effects of calcium and thrombin on growth factor release from platelet concentrates: Kinetics and regulation of endothelial cell proliferation. Biomaterials 25: 4489, 2004 https://doi.org/10.1016/j.biomaterials.2003.11.013