Article
  • Effects of SIS Sponge and Bone Marrow-Derived Stem Cells on the Osteogenic Differentiation for Tissue Engineered Bone
  • Park KS, Jin CM, Yun SJ, Hong KD, Kim SH, Kim MS, Rhee JM, Khang G, Lee HB
  • SIS 스폰지와 골수유래줄기세포를 이용한 조직공학적 골분화 유도
  • 박기숙, 진채문, 윤선중, 홍금덕, 김순희, 김문석, 이종문, 강길선, 이해방
Abstract
Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in 3% (v/v) acetic acid aqueous solution for 48 hrs and freeze-dried. EDC solution (H2O : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100 mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity (ALP), MTT assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation, This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

소장 점막하 조직(SIS)은 면역반응이 없어 생체재료로 널리 사용되고 있다. 본 연구에서는 SIS를 스폰지 형태로 제조하여 1ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC)를 이용하여 경화시켰으며, SIS 스폰지의 구성 원소를 알아보기 위해 원소분석(EA)과 에너지 분산 X선 분광계(EDS)를 사용해 분석하였다. 또한 SIS 함량과 EDC의 농도에 따른 섬유아세포의 부착도 및 성장도를 알아보기 위해 methylthiazoletetrazolium(MTT)을 실시하였다. 이 스폰지에 골수간엽 줄기세포(BMSCs)를 파종해 4주 동안 조직공학적 골분화를 유도하였다. 골분화 유도를 위해 골분화 배지를 사용했으며, 배지에 따른 BMSCs의 세포 성장도와 alkaline phosphatase(ALP) 활성을 측정해 보았다. 또한 역전사 중합연쇄반응을 통해 골분화 여부를 관찰하였다. SEM 관찰 결과 모든 스폰지에서 균일한 형태의 열린 다공이 형성되었음을 확인할 수 있었다. RT-PCR 결과 4주 동안 골분화 배지를 주었을 때 제 I형 교원질이 발현됨을 확인할 수 있었으며, ALP 결과에서도 골분화 배지에서 ALP 활성이 높게 나타남을 볼 수 있엇다. 결론적으로 제조한 SIS 스폰지는 조직공학적 담체로써 우수한 특성을 보이고 있었으며, 또한 BMSCs의 골분화 유도에도 좋은 결과를 보임으로써 조직공학적 골 재생에 잠재적인 가능성을 가지고 있음을 확인할 수 있었다.

Keywords: small intestinal submucosa; sponge; osteogenesis; BMSCs

References
  • 1. Khang G, Lee HBCell-synthetic surface interaction: Physicochemical surface modification, in Methods of Tissue Engineering, A. Atala and R. Lanza, Editors, Academic Press, Section II, pp 771 (2001)
  •  
  • 2. Khang G, Rhee JM, Lee I, Lee HB, Polym. Sci. Technol., 12(4), 527 (2001)
  •  
  • 3. Khang G, Lee I, Lee HB, Lee SJ, Polym. Sci. Technol., 13(1), 4 (2002)
  •  
  • 4. Khang G, Lee SJ, Han CW, Rhee JM, Lee HBPreparation and characterization of natural/synthetic hybrid scaffolds , in Advances in Experimental medicine and Biology, M. Elcin, Editor, Kluwer-Plenum Press, London, Vol. 657, pp 235 (2003)
  •  
  • 5. Khang G, Kim MS, Cho SH, Lee I, Rhee JM, Lee HB, Tissue Eng. Regen. Med., 1, 9 (2004)
  •  
  • 6. Khang G, Lee SJ, Kim MS, Lee HBScaffolds; Tissue Engineering, in Webster's Biomedical Engineering Handbook, S. Webster, Editor, John & Wiley Press, NY, in press (2005)
  •  
  • 7. Habal MB, Clin. Orthop., 188, 239 (1984)
  •  
  • 8. Lane JM, Sandhu HS, Orthop. Clin. North Am., 18, 213 (1997)
  •  
  • 9. Jergesen HE, Chua J, Kao RT, Kaban LB, Clin. Orthop., 268, 253 (1991)
  •  
  • 10. Choi MG, Khang G, Lee IW, Rhee JM, Lee HB, Polym.(Korea), 25(3), 318 (2001)
  •  
  • 11. Jeon EK, Shim JY, Whang HJ, Khang G, Jo I, Lee I, Rhee JM, Lee HB, Biomater. Res., 5, 23 (2001)
  •  
  • 12. Jeon EK, Whang HJ, Khang G, Lee I, Rhee JM, Lee HB, Polym.(Korea), 25(6), 893 (2001)
  •  
  • 13. Kim HS, Lee I, Lee JM, Han CW, Sung JH, Park MY, Khang G, Lee HB, J. Korea Soc. Endorcrinology, 17, 206 (2002)
  •  
  • 14. Lee SJ, Lee DH, Khang G, Lee YM, Lee HB, Macromol. Chem. Symp., 15, 201 (2002)
  •  
  • 15. Jang JW, Lee B, Han CW, Lee IW, Lee HB, Khang G, Polym.(Korea), 27(3), 226 (2003)
  •  
  • 16. Khang G, Park CS, Rhee JM, Lee SJ, Lee YM, Choi MK, Lee HB, Lee I, Korea Polym. J., 9(5), 267 (2001)
  •  
  • 17. Khang G, Rhee JM, Shin P, Kim IY, Lee B, Lee SJ, Lee YM, Lee HB, Lee I, Macromol. Res., 10(3), 158 (2002)
  •  
  • 18. Shin PK, Lee SJ, Lee B, Lee YM, Lee HB, Khang G, Macromol. Chem. Symp., 15, 175 (2002)
  •  
  • 19. Kang JA, Lee SJ, Khang G, Lee I, Lee HB, Biomater. Res., 6, 107 (2002)
  •  
  • 20. Badylak SF, Kropp B, McPherson TB, Tissue Eng., 4, 379 (1998)
  •  
  • 21. Ledet EH, Carl AL, Dirisio DJ, Tymeson MP, Andersen LB, Sheenhan CE, Kallakury B, Slivka M, Serhan H, Spine J., 2, 188 (2002)
  •  
  • 22. Shin HW, Kim SH, Jang JW, Kim MS, Cho SH, Lee HB, Khang G, Polym.(Korea), 28(2), 194 (2004)
  •  
  • 23. Kim SH, Shin HW, Kim MS, Cho SH, Lee HB, Khang G, Biomater. Res., 8, 143 (2004)
  •  
  • 24. Lee SJ, Lee I, Lee YM, Lee HB, Khang G, J. Biomater. Sci.-Polym. Ed., 15, 1003 (2004)
  •  
  • 25. Kim SH, Shin HW, Jang JW, Kim MS, Cho SH, Lee HB, Khang G, Polym.(Korea), 28(6), 478 (2004)
  •  
  • 26. Babylak SF, Lantz GC, Coffey AC, J. Surg. Res., 47, 74 (1989)
  •  
  • 27. Morgan CJ, Coutts AGP, McFadyen MC, King TP, Kelly D, J. Nutr. Biochem., 7, 339 (1996)
  •  
  • 28. Lantz GC, Badylak SF, Coffey AC, J. Invest. Surg., 3, 217 (1990)
  •  
  • 29. Badylak SF, Kropp B, McPherson TB, Tissue Eng., 4, 379 (1998)
  •  
  • 30. Sandusky GE, Badylak SF, Horff RJ, Am. J. Pathol., 140, 317 (1992)
  •  
  • 31. Hong SP, Park JK, Huh SH, Kim HS, J. Plast. Surg., 25, 547 (1998)
  •  
  • 32. Lamme EN, de Vries HJ, van Veen H, Gabbiani G, Westerhof W, Middelkoop E, J. Histochem. Cytochem., 44, 1311 (1996)
  •  
  • 33. Prevel D, Eppley BL, Badylak SF, Ann. Plast. Surg., 35, 381 (1995)
  •  
  • 34. Kropp BP, Eppley BL, Prevel CD, Rippy MK, Harruff RC, Badylak SF, Adams MC, Rink RC, Keating MA, Urology, 46, 396 (1995)
  •  
  • 35. Rosen M, Ponsky J, Petras R, Fanning A, Brody F, Duperier F, Surgery, 132, 480 (2002)
  •  
  • 36. Song SB, Khang G, Hong SK, Lee I, Kim SW, Lee HB, Biomater. Res., 4, 13 (2000)
  •  
  • 37. Park SRCellular interations with cells and extracellular matrix, in Tissue Engineering: Concets and Application, J. J. Yoo and I. Lee, Editors, Korea Medical Book Publisher Co., Seoul, p 43 (2002)
  •  
  • 38. Jang JW, Park KS, Kim SH, Park CS, Kim MS, Han CW, Rhee JM, Khang G, Lee HB, Tissue Eng. Regen. Med., 2, 34 (2005)
  •  
  • 39. Jang JW, Kim EJ, Lee B, Han CW, Kim MS, Cho SH, Lee HB, Khang G, Biomater. Res., 8, 51 (2004)
  •  
  • 40. Kim EJ, Park CS, Kim MS, Cho SH, Rhee JM, Lee HB, Khang G, Polym.(Korea), 28(3), 218 (2004)
  •  
  • Polymer(Korea) 폴리머
  • Frequency : Bimonthly(odd)
    ISSN 0379-153X(Print)
    ISSN 2234-8077(Online)
    Abbr. Polym. Korea
  • 2023 Impact Factor : 0.4
  • Indexed in SCIE

This Article

  • 2005; 29(5): 501-507

    Published online Sep 25, 2005

  • Received on Jul 20, 2005
  • Accepted on Sep 9, 2005