화학공학소재연구정보센터
홈 로그인 로그아웃 연락처 사이트맵
Polymer(Korea), Vol.29, No.5, 501-507, September, 2005
Export Citation
SIS 스폰지와 골수유래줄기세포를 이용한 조직공학적 골분화 유도
Effects of SIS Sponge and Bone Marrow-Derived Stem Cells on the Osteogenic Differentiation for Tissue Engineered Bone
박기숙1, 진채문2, 윤선중2, 홍금덕2, 김순희2, 김문석3, 이종문1, 강길선2, 3, †, 이해방3
  • 1전북대학교 고분자나노공학과
  • 2전북대학교 유기신물질공학과
  • 3한국화학연구원 나노생체재료연구팀
Ki Suk Park1, Chae Moon Jin2, Sun Jung Yun2, Keum Duck Hong2, Soon Hee Kim2, Moon Suk Kim3, John M. Rhee1, Gilson Khang2, 4, †, and Hai Bang Lee3
  • 1Department of PolymerNano Science and Technology, Chonbuk National University, 664-14, Dukjin, Jeonju 561-756, Korea
  • 2Department of Advanced Organic Materials, Chonbuk National University, 664-14, Duckjin, Jeonju 561-756, Korea
  • 3Nanobiomaterials Laboratory, Korea Research Institute of Chemical Technology, P.O. Box 107, Yusung, Daejeon 305-600, Korea
  • 4Nanobiomaterials Laboratory, Korea Research Institute of Chemical Technology
E-mail:
초록
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.
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.
Keywords:small intestinal submucosa;sponge;osteogenesis;BMSCs
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