Department of Pharmacy, The Second Affiliated Hospital of Harbin Medical University, 148 Baojian Road, Harbin, 150086 Heilongjiang Province, China.
Med Biol Eng Comput. 2011 Jun;49(6):687-92. doi: 10.1007/s11517-010-0729-4. Epub 2011 Jan 6.
Injured articular cartilage has a poor capacity for spontaneous healing. So far, a satisfactory solution to repair the injured cartilage has not been found, but transgenic therapy might be a promising treatment. This study aims to evaluate the potential of transfecting bone morphogenetic protein-7 (BMP-7), a secretory protein, into bone marrow-derived mesenchymal stem cells (BMSCs), in inducing the differentiation of bone marrow stromal cells into chondrocytes in vitro. The phenotypes of the cells were observed by alcian blue staining and H&E staining with an inverted microscope. The glycosaminoglycan (GAG) content of BMSCs transfected with pcDNA3.1-BMP7 or induced by inducing medium was examined after 7, 14, or 21 days of incubation. A standard curve as reference for BMSCs' GAG content was plotted using galacturonic acid. The content of type II collagen in culture medium was detected by ELISA. Our results demonstrated that BMP7-transfected BMSCs or BMSCs incubated with inducing medium possess the ability to differentiate into chondrocytes. BMP7-induced BMSCs secrete type II collagen and GAG. There was no significant difference between BMP7-induced BMSCs in their secreted protein content when compared with the positive control group (TGF-β1 and dexamethasone) (P > 0.05), but there was significant difference in the secreted protein profile when compared with the negative control group (P < 0.05).
关节软骨损伤后自我修复能力很差。到目前为止,还没有找到一种令人满意的方法来修复受损的软骨,但转基因治疗可能是一种有前途的治疗方法。本研究旨在评估将分泌蛋白骨形态发生蛋白-7(BMP-7)转染到骨髓间充质干细胞(BMSCs)中,在体外诱导骨髓基质细胞向软骨细胞分化的潜力。通过倒置显微镜观察茜素蓝染色和 H&E 染色的细胞表型。用 galacturonic acid 绘制 BMSCs GAG 含量的标准曲线作为参考,检测 pcDNA3.1-BMP7 转染或诱导培养基诱导 7、14 或 21 天后 BMSCs 的 GAG 含量。通过 ELISA 检测培养上清液中 II 型胶原的含量。结果表明,BMP7 转染的 BMSCs 或用诱导培养基孵育的 BMSCs 具有分化为软骨细胞的能力。BMP7 诱导的 BMSCs 分泌 II 型胶原和 GAG。与阳性对照组(TGF-β1 和地塞米松)相比,BMP7 诱导的 BMSCs 分泌蛋白含量无显著差异(P > 0.05),但与阴性对照组相比,分泌蛋白谱有显著差异(P < 0.05)。