作 者：
阿勒泰别克·闹乎旦;张玉玲;白广超;金洪亮;李宽新
单 位：
新疆兵团医院教学科;新疆兵团医院关节脊柱外科
关键词：
Wnt3a;BMP-7;骨髓间充质干细胞;成骨分化
摘 要：
为了观察Wnt联合BMP诱导骨髓间充质干细胞向成骨方向分化是否具有协同作用,本研究以密度梯度离心联合贴壁法分离培养骨髓间充质干细胞;实验分组为:对照组、Wnt组、BMP-7组、Wnt和BMP-7联合诱导组;以CCK-8法检测细胞的增殖活性;定量测定碱性磷酸酶(ALP)活性;以RT-PCR技术检测特异成骨转录因子的表达情况;使Von Kossa染色法观察细胞内矿化沉积程度。结果显示:以CKK-8法测培养第7天时联合组吸光度值为0.847,Wnt组为0.739,均明显高于对照组0.408,差异具有显著性(P<0.05);联合诱导组的ALP活性比值在培养第6天和第12天时分别为63.9和144.4,BMP-7组分别为40.9和104.2,均明显高于对照组的7.2和18.8,差异具有统计学意义(P<0.05);培养14 d后,OCN、β-Catenin的相对灰度值联合组高于对照组,差异具有统计学意义(P<0.05),而Runx2及Osterix的表达量各组间差异无统计学意义;培养3周后,以Von Kossa染色后观察到钙结节在大小和数量上联合组均大于对照组。由此可知,Wnt3a因子和BMP-7联合可促进骨髓间充质干细胞的成骨分化,且两者具有协同作用。
译 名：
Study on induction of bone marrow mesenchymal stem cells into osteoblasts by combination of Wnt factor and BMP-7
作 者：
Aletaibieke Naohudan;Zhang Yuling;Bai Guangchao;Jin Hongliang;Li Kuanxin;Department of Joint and Spinal,Xinjiang Production and Construction Corps Hospital;Department of Science and Education,Xinjiang Production and Construction Corps Hospital;
关键词：
Wnt3a;;BMP-7;;marrow mesenchymal stem cells;;osteogenesis
摘 要：
To observe the synergistic effect of Wnt combined with BMP on osteogenic differentiation of bone marrow mesenchymal stem cells,isolated culture was performed on MSCs by density-gradient centrifugation combined with adherence method.The control group,Wnt group,BMP-7 group,Wnt-BMP-7 combined induction group were defined.The cell proliferative activity was detected by CCK-8 method.The ALP activity was detected quantitatively.The expression levels of the osteoblast-specific transcription factors were detected by RT-PCR.The degree of intracellular mineral deposition was observed by Von Kossa staining method.The results showed that the light absorbance at 7 days of culture with CKK-8 method was 0.847 in the combined induction group and 0.739 in the Wnt group,both of which were significantly higher than 0.408 in the control group(P<0.05);the ALP activity ratios at 6 and 12 days of culture was 63.9 and 144.4,respectively in the combined induction group,and 40.9 and 104.2,respectively in the BMP-7 group,which were all significantly higher than 7.2 and 18.8,respectively in the control group(P<0.05); at 14 days,the relative gray values of OCN and β-Catenin were higher in the combined induction group than in the control group(P<0.05),while there was no statistically significant difference in the expression levels of Runx2 and Osterix across the groups; at 3 weeks of culture,both the size and number of calcium nodules observed following Von Kossa staining were greater in the combined induction group than in the control group.The conclusion is that combination of Wnt3 a and BMP-7 has synergistic action in promoting differentiation of MSCs into osteoblasts.
