去卵巢骨质疏松大鼠骨髓间充质干细胞的体外成骨诱导

背景：细胞学研究表明,骨髓间充质干细胞在绝经后骨质疏松症的发病过程中起有重要作用。目的：观察去卵巢骨质疏松大鼠骨髓间充质干细胞的体外成骨分化。 方法：将6月龄雌性SD大鼠双侧卵巢切除建立骨质疏松模型。实验分为4组：正常干细胞组、骨质疏松干细胞组、正常干细胞成骨诱导组、骨质疏松干细胞成骨诱导组。全骨髓贴壁法培养正常和骨质疏松大鼠骨髓间充质干细胞至第3代细胞用于实验。倒置相差显微镜观察细胞形态,流式细胞仪测定细胞周期、增殖指数。加成骨诱导液进行成骨诱导,检测各组骨髓间充质干细胞碱性磷酸酶活性,茜素红染色法比较各组钙结节的形成。 结果与结论：正常干细胞成骨诱导组、骨质疏松干细胞成骨诱导组均具有成骨细胞的形态特征,但骨质疏松干细胞成骨诱导组形态变化相对缓慢。正常干细胞组细胞增殖指数高于骨质疏松干细胞组(P < 0.05);成骨诱导组碱性磷酸酶活性均明显高于相应的正常或骨质疏松干细胞组(P < 0.05);正常干细胞成骨诱导组明显高于骨质疏松干细胞成骨诱导组(P < 0.05)。成骨诱导组茜素红染色均呈阳性,相应的正常或骨质疏松干细胞组呈阴性;且正常干细胞成骨诱导组染色强于骨质疏松干细胞成骨诱导组。提示去卵巢骨质疏松大鼠骨髓间充质干细胞的增殖能力和成骨分化能力明显降低,可能与去卵巢大鼠骨质疏松的发生相关。

In vitro osteogenic differentiation of bone marrow mesenchymal stem cells from ovariectomied osteoporotic rats

BACKGROUND: Cytological studies show that bone marrow mesenchymal stem cells play an important role in postmenopausal osteoporosis mechanism. OBJECTIVE: To study the osteogenic differentiation in vitro of bone marrow mesenchymal stem cells from ovariectomied osteoporotic rats.  METHODS: The osteoporotic animal model was established by performing ovariectomy in the 6-month-old female Sprague-Dawley rats. There were four groups: bone marrow mesenchymal stem cells control group, bone marrow mesenchymal stem cells osteoporosis group, bone marrow mesenchymal stem cells osteogenic induction group and oseogenesis induction group. Bone marrow mesenchymal stem cells were isolated from the rats of control group and oseogenesis induction group by means of the whole bone marrow adherence method and cultured to the 3rd generation. Then the bone marrow mesenchymal stem cells were used in all the experiments. Cell morphology was observed under the inverted phase contrast microscope, cell cycle and proliferation index of bone marrow mesenchymal stem cells were detected by flow cytometry. After osteogenic induction, the expression level of alkaline phosphatase was detected, and the fornation of calcium nodes of bone marrow mesenchymal stem cells were marked by alizarin red staining.RESULTS AND CONCLUSION: The cells in the osteogenic induction group and oseogenesis induction group had the morphology of osteobalsts, and the change of morphology of the cells in the oseogenesis induction group was relatively tardiness. The proliferation index in the control group was higher than that in the osteoporosis group (P < 0.05); expression level of alkaline phosphatase in the osteogenic induction group was significantly higher than that in the oseogenesis induction group (P < 0. 05), and the control group was significantly higher than the oseogenesis group (P < 0.05). The alizarin red staining of the cells in the osteogenic induction group was positive, while negative in the control group and the oseogenesis group; the staining in the osteogenic induction group was stronger than that in the oseogenesis induction group. These findings indicate that both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cells from the ovariectomized osteoporotic rats are decreased, which may be related with the ostoeporosis pathogensis of ovariectomied rats.