Liao Junyi, Zhou Nian, Lin Liangbo, Yi Shixiong, Fan Tingxu, Zhao Chen, Hu Ning, Liang Xi, Si Weike, Huang Wei
Department of Orthopedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2014 Mar;34(3):317-22.
To investigate the effect of co-expression of bone morphogenetic protein 2 (BMP2) and Sox9 on chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro and provide experimental evidence for tissue engineering of cartilage.
Mouse embryonic bone marrow MSC C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP2, Sox9 and green fluorescent protein (GFP) for 3-14 days, with cells infected with the adenovirus carrying GFP gene as the control. The mRNA expression of the markers of chondrogenic differentiation, including collagen type II (Col2a1), aggrecan (ACAN), and collagen type X (Col10a1), were determined by real-time PCR. Alcian blue staining was used for quantitative analysis of sulfated glycosaminoglycan in the cellular matrix. The expression of Col2a1 protein was assayed by immunohistochemical staining and Western blot analysis.
Adenovirus-mediated BMP2 expression induced chondrogenic differentiation of C3H10T1/2 cells. Overexpression of Sox9 effectively enhanced BMP2-induced expression of the chondrogenic markers Col2a1, aggrecan and Col10a1 mRNAs, and promoted the synthesis of sulfated glycosaminoglycan and Col2a1 protein in C3H10T1/2 cells.
Co-expression of BMP2 and Sox9 can promote chondrogenic differentiation of MSCs in vitro, which provides a new strategy for tissue engineering of cartilage.
探讨骨形态发生蛋白2（BMP2）与Sox9共表达对间充质干细胞（MSCs）体外软骨分化的影响，为软骨组织工程提供实验依据。
用表达BMP2、Sox9和绿色荧光蛋白（GFP）的重组腺病毒感染小鼠胚胎骨髓MSCs C3H10T1/2细胞3 - 14天，以感染携带GFP基因腺病毒的细胞作为对照。通过实时聚合酶链反应（PCR）检测软骨分化标志物Ⅱ型胶原（Col2a1）、聚集蛋白聚糖（ACAN）和Ⅹ型胶原（Col10a1）的mRNA表达。采用阿尔新蓝染色对细胞基质中的硫酸化糖胺聚糖进行定量分析。通过免疫组织化学染色和蛋白质印迹分析检测Col2a1蛋白的表达。
腺病毒介导的BMP2表达诱导C3H10T1/2细胞发生软骨分化。Sox9的过表达有效增强了BMP2诱导的软骨分化标志物Col2a1、聚集蛋白聚糖和Col10a1 mRNA的表达，并促进了C3H10T1/2细胞中硫酸化糖胺聚糖和Col2a1蛋白的合成。
BMP2与Sox9共表达可促进MSCs体外软骨分化，为软骨组织工程提供了新策略。
