摘要
目的探讨含2型胶原蛋白(Col2)、硫酸软骨素(CS)和(或)透明质酸(HA)的软骨仿生基质材料,在体外条件下对大鼠骨髓间充质干细胞(BMSC)分化为软骨细胞样细胞的影响。方法体外培养BMSC,分别接种于对照材料、含Col2、Col2-CS、Col2-HA、Col2-CS-HA的软骨仿生基质材料。诱导7、14、21、28 d后,采用CCK-8法检测各复合基质对BMSC增殖、分化活性的影响;采用反转录PCR技术检测软骨细胞标志物Col2、Y染色体性别决定区相关高迁移率组基因9(SOX9)及聚集蛋白聚糖(aggrecan)的mRNA水平,免疫荧光技术检测Col2、SOX9及aggrecan的蛋白水平。结果 CCK-8法检测结果显示,以Col2为基础的基质材料都能作为BMSC生长的良好细胞外基质,CS和HA使BMSC的增殖活性逐渐增加;各复合基质材料均能在体外有效诱导BMSC向软骨细胞样细胞分化;Col2-CS-HA包被能够最大程度上调Col2、SOX9及aggrecan mRNA及蛋白的水平。结论以Col2为基础的仿生基质材料可在体外有效诱导大鼠BMSC分化为软骨细胞样细胞,其中以复合Col2-CS-HA基质材料效率最高。
Objective To investigate the effect of chondrocytic biomimetic matrix scaffold materials containing type 2 collagen （Col2） hydrogel, chondroitin sulphate （CS） and （or） hyaluronic acid （HA） on chondrogenic differentiation of rat bone marrow mesenchymal stem cells （BMSCs） in vitro. Methods BMSCs were cultured in different groups as following： control, Col2, Col2-CS, Col2-HA, Col2-CS-HA. On day 7, 14, 21 and 28, cell proliferation and differentiation activities were determined using CCK-8 assay. The mRNA and protein expressions of Col2, sex-determining region Y （SRY）-related high mobility group-box gene 9 （SOXg） and aggrecan were detected using reverse transcription PCR and immunofluorescence technique. Results CCK-8 assay indicated that chondrocytic biomimetic matrix scaffold materials based on Col2 could be used as good extracellular matrix for BMSCs culture in vitro. CS and HA increased the proliferation and differentiation activities. All the groups induced effectively BMSCs to differentiate into chondrocytes. Compared with the other groups, Col2-CS-HA-coated group up-regulated the mRNA and protein expressions of Col2, SOX9 and aggrecan the most effectively. Conclusion Chondrocytic biomimetic matrix scaffold materials based on Col2 could effectively induce rat BMSCs to differentiate into chondrocyte-like cells in vitro, and supplementation of the hydrogels with both CS and HA had the maximum efficacy.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第11期1502-1505,1510,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81271981)
关键词
仿生基质
骨髓间充质干细胞
软骨细胞样细胞
extracellular matrix
bone marrow mesenchymal stem cells
chondrecyte-like cells
