Polymer(Korea) (폴리머)

The Polymer Society of Korea (한국고분자학회)
권호 표지

Effects of SIS Sponge and Bone Marrow-Derived Stem Cells on the Osteogenic Differentiation for Tissue Engineered Bone

SIS 스폰지와 골수유래줄기세포를 이용한 조직공학적 골분화 유도

  • Park Ki Suk (Department of Polymer Nano Science and Technology, Chonbuk National University) ;
  • Jin Chae Moon (Department of Advanced Organic Materials, Chonbuk National University) ;
  • Yun Sun Jung (Department of Advanced Organic Materials, Chonbuk National University) ;
  • Hong Keum Duck (Department of Advanced Organic Materials, Chonbuk National University) ;
  • Kim Soon Hee (Department of Advanced Organic Materials, Chonbuk National University) ;
  • Kim Moon Suk (Nanobiomaterials Laboratory, Korea Research Institute of Chemical Technology) ;
  • Rhee John M. (Department of Polymer Nano Science and Technology, Chonbuk National University) ;
  • Khang Gilson (Department of Advanced Organic Materials, Chonbuk National University, Nanobiomaterials Laboratory, Korea Research Institute of Chemical Technology) ;
  • Lee Hai Bang (Nanobiomaterials Laboratory, Korea Research Institute of Chemical Technology)
  • 박기숙 (전북대학교 고분자나노공학과) ;
  • 진채문 (전북대학교 유기신물질공학과) ;
  • 윤선중 (전북대학교 유기신물질공학과) ;
  • 홍금덕 (전북대학교 유기신물질공학과) ;
  • 김순희 (전북대학교 유기신물질공학과) ;
  • 김문석 (한국화학연구원 나노생체재료연구팀) ;
  • 이종문 (전북대학교 고분자나노공학과) ;
  • 강길선 (전북대학교 유기신물질공학과, 한국화학연구원 나노생체재료연구팀) ;
  • 이해방 (한국화학연구원 나노생체재료연구팀)
  • Published : 2005.09.01
Download PDF
⟨ Previous Next ⟩

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 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. 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), n 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 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride(EDC)를 이용하여 경화시켰으며, SIS 스폰지의 구성 윈소를 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 골수 간엽 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 ALP활성이 높게 나타남을 볼 수 있었다. 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

References

  1. G. Khang and H. B. Lee, 'Cell-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. G. Khang, I. Lee, J. M. Rhee, and H. B. Lee, Polymer Science & Technology, 12,527 (2001)
  3. G. Khang, S. J. Lee, I. Lee, and H. B. Lee, Polymer Science & Technology, 13, 4 (2002)
  4. G. Khang, S. J. Lee, C. W. Han, J. M. Rhee, and H. B. Lee, 'Preparation and characterization of natural/synthetic hybrid scaffolds', in Advances in Experimental Medicine and Biology, M. Elcin, Editor, KIuwer-Plenum Press, London, Vol. 657, pp 235 (2003)
  5. G. Khang, M. S. Kim, S. H. Cho, I. Lee, J. M Rhee, and H. B. Lee, Tissue Eng. Regen. Med., 1, 9 (2004)
  6. G. Khang, S. J. Lee, M. S. Kim, and H. B. Lee, 'Scaffolds., Tissue Engineering', in Webster's Biomedical Engineering Handbook, S. Webster, Editor, John & Wiley Press, NY, in press, 2005
  7. M. B. Habal, Clin. Orthop., 188, 239 (1984)
  8. J. M. Lane and H. S. Sandhu, Orthop. Clin. North Am., 18, 213 (1997)
  9. H. E. Jergesen, J. Chua, R. T. Kao, and L. B. Kaban, Clin. Orthop., 268, 253 (1991)
  10. M. K. Choi, G. Khang, I. Lee, J. M. Rhee, and H. B. Lee, Polymer(Korea), 25, 318 (2001)
  11. E. K. Jeon, J. Y. Shim, H. J. Whang, G. Khang, I. Jo, I. Lee, J. M. Rhee, and H. B. Lee, Biomater. Res., 5, 23 (2001)
  12. E. K. Jeon, G. Khang, I. Lee, J. M. Rhee, and H. B. Lee, Polymer(Korea), 25, 893 (2001)
  13. H. S. Kim, I. Lee, J. M. Lee, C. W. Han, J. H. Sung, M. Y. Park, G. Khang, and H. B. Lee, J. Korea Soc. Endorcrinology, 17, 206 (2002)
  14. S. J. Lee, D. H. Lee, G. Khang, Y. M. Lee, and H. B. Lee, Macromol. Chem. Symp., 15, 201 (2002)
  15. J. W. Jang, B. Lee, C. W. Han, I. Lee, H. B. Lee, and G. Khang, Polymer(Korea), 27, 226 (2003)
  16. G. Khang, C. S. Park, J. M. Rhee, S. J. Lee, Y. M. Lee, I. Lee, M. K. Choi, and H. B. Lee, Korea Polym. J., 9, 267 (2001)
  17. G. Khang, P. Shin, I. Kim, B. Lee, S. J. Lee, Y. M. Lee, H. B. Lee, and I. Lee, Macromol. Res., 10, 158 (2002) https://doi.org/10.1007/BF03218266
  18. P. K. Shin, S. J. Lee, B. Lee, Y. M. Lee, H. B. Lee, and G. Khang, Macromol. Chem. Symp., 15, 175 (2002)
  19. J. A. Kang, S. J. Lee, G. Khang, I. Lee, and H. B. Lee, Biomater. Res., 6, 107 (2002)
  20. S. F. Badylak, B. Kropp, and T. B. McPherson, Tissue Eng., 4, 379 (1998) https://doi.org/10.1089/ten.1998.4.379
  21. E. H. Ledet, A. L. Carl, D. J. Dirisio, M. P. Tymeson, L. B. Andersen, C. E. Sheenban, B. Kallakury, M. Slivka, and H. Seman, Spine J., 2, 188 (2002) https://doi.org/10.1016/S1529-9430(02)00182-1
  22. H. W. Shin, S. H. Kim, J. W. Jang, M. S. Kim, S. H. Cho, H. B. Lee, and G. Khang, Polymer(Korea), 28, 194 (2004)
  23. S. H. Kim, H. W. Shin, M. S. Kim, S. H. Cho, H. B. Lee and G. Khang, Biomater. Res., 8, 143(2004)
  24. S. J. Lee, I. Lee, Y. M. Lee, H. B. Lee, and G. Khang, J. Biomater. Sci., Polym. Ed., 15, 1003(2004) https://doi.org/10.1163/1568562041526487
  25. S. H. Kim, H. W. Shin, J. W. Jang, M. S. Kim, S. H. Cho, H. B. Lee, and G. Khang, Polymer(Korea), 28, 478 (2004)
  26. S. F. Badylak, G. C. Lantz, and A. C. Coffey, J. Surg. Res., 47, 74 (1989) https://doi.org/10.1016/0022-4804(89)90050-4
  27. C. J. Morgan, A. G. P. Coutts, M. C. McFadyen, T. P. King, and D. Kelly, J. Nutri. Biochem., 7, 339 (1996) https://doi.org/10.1016/0955-2863(96)00054-X
  28. G. C. Lantz, S. F. Badylak, and A. C. Coffey, J. Invest. Surg., 3, 217 (1990) https://doi.org/10.3109/08941939009140351
  29. S. F. Badylak, B. Kropp, and T. B. McPherson, Tissue Eng., 4, 379 (1998) https://doi.org/10.1089/ten.1998.4.379
  30. G. E. Sandusky, S. F. Badylak, and R. J. Horff, Ann. J. Path., 140, 317 (1992)
  31. S. P. Hong, J. K. Park, S. H. Huh, and H. S. Kim, J. Plast. Surg., 25, 547 (1998)
  32. E. N. Lamme, H. J. de Vries, H. van Veen, G. Gabbiani, W. Westerhof, and E. Middelkoop, J. Histochem. Cytochem., 44, 1311 (1996) https://doi.org/10.1177/44.11.8918906
  33. D. Prevel, B. L. Eppley, and S. F. Badylak, Ann. Plast. Surg., 35, 381 (1995) https://doi.org/10.1097/00000637-199510000-00009
  34. B. P. Kropp, B. L. Eppley, C. D. Prevel, M. K. Rippy, R. C. Harruff, S. F. Badylak, M. C. Adams, R. C. Rink, and M. A. Keating, Urology, 46, 396 (1995) https://doi.org/10.1016/S0090-4295(99)80227-1
  35. M. Rosen, J. Ponsky, R. Petras, A. Fanning, F. Brody, and F. Duperier, Surgery, 132, 480 (2002) https://doi.org/10.1067/msy.2002.126505
  36. S. B. Song, G. Khang, S. K. Hong, I. Lee, S. W. Kim, and H. B. Lee, Biomater. Res., 4, 13 (2000)
  37. S. R. Park, 'Cellular interations with cells and extracellular matrix', in Tissue Engineering: Concepts and Application, J. J. Yoo and I. Lee, Editors, Korea Medical Book Publisher Co., Seoul, p 43 (2002)
  38. J. W. Jang, K. S. Park, S. H. Kim, C. S. Park, M. S. Kim, C. W. Han, J. M. Rhee, G. Khang and H. B. Lee, Tissue Eng. Regen. Med., 2, 34 (2005)
  39. J. W. Jang, E. J. Kim, B. Lee, C. W. Han, M. S. Kim, S. H. Cho, H. B. Lee, and G. Khang, Biomater. Res. 8, 51 (2004)
  40. E. J. Kim, J. W. Jang, M. S. Kim, S. H. Cho, H. B. Lee, and G. Khang, Polymer(Korea), 28, 218 (2004)