Optimal Condition of Microporous Membrane for Bone Marrow Stromal Cell Allotransplantation to Stimulate Wound Healing in Vitro

창상치유목적의 골수기질세포 동종이식을 위한 고분자막의 조건

  • Lee, Eun-Sang (Department of Plastic Surgery, Korea University College of Medicine) ;
  • Kim, Myeong-Joo (Department of Plastic Surgery, Korea University College of Medicine) ;
  • Han, Seung-Kyu (Department of Plastic Surgery, Korea University College of Medicine) ;
  • Hong, Sung-Taek (Department of Plastic Surgery, Korea University College of Medicine) ;
  • Kim, Woo-Kyung (Department of Plastic Surgery, Korea University College of Medicine)
  • 이은상 (고려대학교 의과대학 성형외과학교실) ;
  • 김명주 (고려대학교 의과대학 성형외과학교실) ;
  • 한승규 (고려대학교 의과대학 성형외과학교실) ;
  • 홍성택 (고려대학교 의과대학 성형외과학교실) ;
  • 김우경 (고려대학교 의과대학 성형외과학교실)
  • Received : 2010.05.17
  • Accepted : 2010.06.22
  • Published : 2010.09.10

Abstract

Purpose: Major drawbacks of conventional bone marrow stromal cells (BSCs) transplantation method are mainly caused by direct transplanted cell to host cell interactions. We hypothesized that separation of the transplanted cells by a microporous membrane might inhibit most of the potential adverse effects and induce superior effect. The purpose of the study is to determine the optimal condition of the microporous membrane. Methods: First, BSCs were placed in polyethylene terephthalate (PET) transwell inserts with 3, 8, or $12{\mu}m$ pore size, and cultured in 24 well culture plates. After 5 days, bottoms of the plates were observed for presence of attached BSCs in monolayer and cell numbers were evaluated. Second, BSCs were placed PET, polycarbonate (PCT), and mixed cellulose esters (MCE) transwell inserts with 3 and $8{\mu}m$ pore size, and cultured in 24 well culture plates. After 3 days, the supernatants of the media left in culture plate were analyzed for collagen, vascular endothelial growth factor (VEGF), platelet derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF). Third, BSCs were placed in 15% and 70% of the PET membrane with $3{\mu}m$ pore size. All the experimental conditions and methods were same as the second study. Results: The optimal pore sizes to prevent BSC leakage were $3{\mu}m$ and $8{\mu}m$. The amounts of type I collagen and three growth factors tested did not show significant differences among PET, PCT, and MCE groups. However, the collagen, VEGF, and bFGF levels were much higher in the high (70%) density group than in the low (15%) density group. Conclusion: This study revealed that the optimal pore size of membrane to prevent direct BSC to recipient cell contact is in between $3{\mu}m$ and $8{\mu}m$. Membrane materials and pore sizes do not influence the collagen and growth factor passage through the membrane. The most striking factor for collagen and growth factor transport is pore density of the membrane.

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. Kuroyanagi Y, Yamada N, Yamashita R, Uchinuma E: Tissue-engineered product: allogenic cultured dermal substitute composed of spongy collagen with fibroblasts. Artif organs 25: 180, 2001 https://doi.org/10.1046/j.1525-1594.2001.025003180.x
  3. Mansbridge JN, Liu K, Pinney RE, Patch R, Ratcliffe A, Naughton GK: Growth factors secreted by fibroblast: role in healing diabetic foot ulcers. Diabetes Obes Metab 1: 265, 1999 https://doi.org/10.1046/j.1463-1326.1999.00032.x
  4. Han SK, Yoon TH, Lee DG, Lee MA, Kim WK: Potential of human bone marrow stromal cells to accelerate wound healing in vitro. Ann Plast Surg 55: 414, 2005 https://doi.org/10.1097/01.sap.0000178809.01289.10
  5. Han SK, Chun KW, Gye MS, Kim WK: The effect of human bone marrow stromal cells and dermal fibroblasts on angiogenesis. Plast Reconstr Surg 117: 829, 2006 https://doi.org/10.1097/01.prs.0000201458.80364.31
  6. Embil JM, Papp K, Sibbald G, Tousignant J, Smiell JM, Wong B, Lau CY: Recombinant human platelet-derived growth factor-BB (becaplermin) for healing chronic lower extremity diabetic ulcers: an open-label clinical evaluation of efficacy. Wound Repair Regen 8: 162, 2000 https://doi.org/10.1046/j.1524-475x.2000.00162.x
  7. Tsang MW, Wong WK, Hung CS, Lai KM, Tang W, Cheung EY, Kam G, Leung L, Chan CW, Chu cm, Lam EK: Human epidermal growth factor enhances healing of diabetic foot ulcers. Diabetes Care 26: 1856, 2003 https://doi.org/10.2337/diacare.26.6.1856
  8. Conget PA, Minguell JJ: Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol 181: 67, 1999 https://doi.org/10.1002/(SICI)1097-4652(199910)181:1<67::AID-JCP7>3.0.CO;2-C
  9. Watanabe N, Woo SL, Papageorgiou C, Celechovsky C, Takai S: Fate of donor bone marrow cells in medial collateral ligament after simulated autologous transplantation. Microsc Res Tech 58: 39, 2002 https://doi.org/10.1002/jemt.10115
  10. Quirici N, Soligo D, Bossolasco P, Servida F, Lumini C, Deliliers GL: Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies. Exp Hematol 30: 783, 2002 https://doi.org/10.1016/S0301-472X(02)00812-3
  11. Prockop DJ: Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276: 71, 1997 https://doi.org/10.1126/science.276.5309.71
  12. Briganti E, Losi P, Raffi A, Scoccianti M, Munao A, Soldani G: Silicone based polyurethane materials: a promising biocompatible elastomeric formulation for cardiovascular applications. J Mater Sci Mater Med 17: 259, 2006 https://doi.org/10.1007/s10856-006-7312-4
  13. Gerald K: Introduction to the study of cell biology. Cell and molecular biology. 1st ed, New York, John Wiley & Sons, 1996, p 17-19
  14. Ng KW, Leong DT, Hutmacher DW: The challenge to measure cell proliferation in two and three dimensions. Tissue Eng 11: 182, 2005 https://doi.org/10.1089/ten.2005.11.182
  15. Striz I, Slavcev A, Kalanin J, Jaresova M, Rennard SI: Cell-cell contacts with epithelial cells modulate the pheno-type of human macrophages. Inflammation 25: 241, 2001 https://doi.org/10.1023/A:1010975804179