基底膜基质/壳聚糖诱导骨髓间充质干细胞成软骨分化  

Chondrogenic differentiation of mesenchymal stem cells on matrigel/chitosan scaffolds

作　　者：戴云[1] 何静[1] 吴方[1] 

机构地区：[1]四川大学生物材料工程研究中心,四川省成都市610064

出　　处：《中国组织工程研究》2015年第10期1506-1510,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助(81271702)~~

摘　　要：背景:课题组前期研究发现基底膜基质能够定向诱导骨髓间充质干细胞向软骨方向分化,但其力学性能与实际应用有较大差距,还需要进一步研究。目的:制备兼具适宜力学性能和优异生物相容性的壳聚糖/基底膜基质水凝胶支架用于软骨修复。方法:以京尼平为交联剂,将壳聚糖溶液与基底膜基质按2︰1,1︰1,1︰3比例混合制成水凝胶,接种大鼠骨髓间充质干细胞,培养14 d。经材料力学测试、细胞增殖、活细胞染色、酶联免疫吸附测试以及阿尔新蓝染色等方法评价材料诱导细胞成软骨分化能力。结果与结论:在基底膜基质内添加壳聚糖后,材料力学性能从0.48 k Pa上升到1.78 k Pa。标志性蛋白分泌结果显示,纯壳聚糖组早期诱导成软骨活性高于其他组,但后期诱导能力减弱,而含基底膜基质各组在后期能够保持较好的诱导活性,其中壳聚糖/基底膜基质=1︰1组材料具有一定的力学强度,且Ⅱ型胶原和Ⅹ型胶原的表达量在14 d较其他组高。结果表明实验制备的壳聚糖/基底膜基质水凝胶具有良好的力学性能,并能够促进骨髓间充质干细胞向软骨方向分化,可用于软骨组织工程研究。BACKGROUND： Previous studies have found that the basement membrane matrix can induced the chondrogenic differentiation of bone marrow mesenchymal stem cells, but there is a wide gap between its mechanical properties and practical application, and further research is needed.OBJECTIVE： To prepare a suitable matrigel/chitosan scaffold that has appropriate mechanical properties and remarkable bioactivity for cartilage repair.METHODS： We selected genipin as cross-linking agent, and mixed Matrigel with cross-linked chitosan at different ratios（2：1, 1：1, 1：3）. Then rat bone marrow mesenchymal stem cells were seeded on different scaffolds and cultured for 14 days. The mechanical properties of materials were measured by DMA. Cell counting kit-8, FDA staining, ELISA kits and Alcian blue staining were used to measure the bioactivity of materials.RESULTS AND CONCLUSION： The storage modulus of scaffolds was raised from 0.48 k Pa to 1.78 k Pa with increase ratio of chitosan. Cells spread well in the early period on all scaffolds, and then the cells on the chitosan scaffold showed reduced chondrogenic activity, but cells on the scaffolds with matrigel could maintain chondrogenic differentiation. The matrigel/chitosan scaffold at a ratio of 1：1 had appropriate mechanical properties and higher levels of collagen II and collagen X at 14 days. The prepared matrigel/chitosan scaffold with decent mechanical performance can promote the differentiation of bone marrow mesenchymal stem cells into chondrogenic lineages, which can be used in cartilage tissue engineering.

关 键 词：骨髓 间质干细胞 基底膜 壳聚糖 干细胞 骨髓干细胞 基底膜基质 骨髓间充质干细胞 力学性能 成软骨分化 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]