摘要
目的探索明胶-硫酸软骨素-透明质酸钠多孔载体作为新型组织工程软骨载体的可行性。方法观察-80℃冷冻抽真空干燥制备的明胶-硫酸软骨素-透明质酸钠载体的孔径、孔隙率、交通孔和密度;分离、扩增兔骨髓基质干细胞(MSCs),MTT检测MSCs在载体上的黏附率及多孔载体对细胞增殖功能的影响;兔MSCs经转化生长因子β1(TGF-β1)诱导7d后免疫组化检测Ⅱ型胶原、S-100蛋白表达;诱导后的MSCs与载体复合后植入裸鼠肌袋内,分别于术后2、4周取材进行组织学观察。结果载体孔径为230±30μm,孔隙率为79%,密度为11.50±0.36(μg/mm3);细胞在载体上黏附率87.5%;载体可轻度促进MSCs增殖;经TGF-β1诱导7d后MSCsⅡ型胶原、S-100蛋白免疫组化染色阳性;诱导后的MSCs在体内生长增殖良好,4周时可见新生软骨组织形成。结论该载体具有良好的孔径、孔隙率,与MSCs具有较好的相容性,是软骨组织工程中的一种新型仿生载体。
Objective To investigate the feasibility of using the gelatin- chondroitin sulfate- sodium hyaluronate tri- copolymer scaffold as a novel cartilage tissue engineering scaffold. Methods The scaffold was prepared with gelatin - chondroitin sulfate- sodium hyaluronate tri - copolymer at - 80℃. Pore size, porosity, internal pores and density were observed with light microscopy and scanning electron microscopy （SEM）. The number of bone marrow mesenchymal stem cells （MSCs） attached to scaffold and the function of cells were also determined with MTT colorimetric microassay. MSCs were cultured in the medium of DMEM with TGFβ1 microsphere for 7 days. Immunohistochemistry was used for detecting the expression of type 11 collagen and S- 100 protein. Scaffold was combined with induced MSCs and then transplanted into the muscle of nude mice. The mice were sacrificed after 2 and 4 weeks respectively and samples were prepared for histological examination. Results The pore size, porosity, and density were 230 ± 30μ, 79% and 11.50 ± 0.36μg/mm^3 respectively. The cell adhesion rate was 87.5 %. The scaffold could promote the proliferation of MSCs mildly. The differentiation of MSCs toward chondrogenic phenotypc was verified by the positive result of type Ⅱ collagen and S- 100 protein in cells cultured in the medium of DMEM with TGFβ1 microsphere. The induced MSCs were proliferated successfully and new cartilage was found by the end of 4 weeks. Conclusion The gelatin- chondroitin sulfate- sodium hyaluronate tri - copolymer scaffold has proper pore size, porosity and internal pores. It is a novel and potential scaffold in cartilage tissue engineering.
出处
《中国骨与关节损伤杂志》
2006年第1期33-35,共3页
Chinese Journal of Bone and Joint Injury
基金
国家"九五"高技术计划新材料领域基金资助项目(009-0160)
