BMP-12促进PLGA/明胶材料培养的人骨髓间充质干细胞增殖和成肌腱分化  

BMP-12 Promotes Proliferation and Tenogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Cultured on PLGA/Gelatin Material

作　　者：姜杨[1] 王璐璐 侯继野 张晓东[1] 徐桂清[1] 王玉 沈雷[1,2] 吴雨轩 Jiang Yang;Wang Lulu;Hou Jiye;Zhang Xiaodong;Xu Guiqing;Wang Yu;Shen Lei;Wu Yuxuan(Department of Anatomy,Qiqihar Medical U-niversity,Qiqihar Heilongjiang 161006,China;不详)

机构地区：[1]齐齐哈尔医学院解剖教研室,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院基础医学院基础医学研究所,黑龙江齐齐哈尔161006 [3]齐齐哈尔市建华医院介入科,黑龙江齐齐哈尔161006 [4]齐齐哈尔医学院附属第三医院内分泌科,黑龙江齐齐哈尔161006

出　　处：《航天医学与医学工程》2020年第5期390-396,共7页Space Medicine & Medical Engineering

基　　金：国家自然科学基金(81541137);齐齐哈尔医学院科研基金项目(QY2016M-03,QY2017Z-01,QMSI2019M-06)。

摘　　要：目的探讨三维培养条件下,骨形态发生蛋白-12(BMP-12)对人骨髓间充质干细胞(hBMSC)增殖和成肌腱分化的影响。方法静电纺丝技术制备PLGA/明胶材料。hBMSCs分为6组:对照组(Con),细胞正常培养;BMP-12组,细胞中添加10ng/mL BMP-12培养;Triciribine组,BMP-12组细胞添加5μmol/L Triciribine;材料组(Material),hBMSCs种植在PLGA/明胶材料;3D-BMP-12组,材料组中添加10ng/mL BMP-12;3D-Triciribine组,3D-BMP-12组添加5μmol/L Triciribine。各组hBMSCs均进行成肌腱诱导分化。扫描电子显微镜检测材料结构;CCK-8法检测各组细胞增殖;Western blot检测各组Scleraxis(Scx)、Tenomodulin(TNMD)和Tenascin-R、Akt、p-Akt蛋白表达。结果与对照组相比,材料组、BMP-12组细胞生长数目均明显升高。与材料组相比,3D-BMP-12组hBMSCs细胞数量、Scx、TNMD和Tenascin-R蛋白表达均明显升高(P<0.01),3D-Triciribine组hBMSCs上述检测指标均分别比3D-BMP-12组显著降低(P<0.01)。3D-BMP-12组hBMSCs的Akt、p-Akt蛋白表达均显著升高,3D-Triciribine组Akt、p-Akt蛋白表达均明显降低(P<0.01)。结论 BMP-12能激活Akt信号通路,促进生物材料上hBMSCs增殖和成肌腱分化。Objective To investigate the effect of bone morphogenetic protein-12(BMP-12)on the proliferation and tenogenic differentiation of human bone marrow mesenchymal stem cells(hBMSC)under three-dimensional culture conditions.Methods PLGA/gelatin materials were prepared by electrospinning technology.hBMSCs were divided into 6 groups:Control group(Con),incubated under conventional culture condition;BMP-12 group,treated with 10 ng/mL BMP-12;Triciribine group,adding 5μmol/L Triciribine to BMP-12 group;Material group(Material),hBMSCs seeded in PLGA/gelatin materials;3D-BMP-12 group,adding 10 ng/mL BMP-12 to Material group;3D-Triciribine group,adding 5μmol/L Triciribine to 3D-BMP-12 group.The hBMSCs in each group were induced to differentiate into tenocytes.Material structures were observed by scanning electron microscope and cell proliferation was detected by CCK-8 assay.Western blot was applied to explore the expression of Scleraxis(Scx),Tenomodulin(TNMD),Tenascin-R,Akt and p-Akt.Results The number of hBMSCs in the material group and the BMP-12 group increased sequentially as compared with the Control group.The relative absorbance and expressions of Scx,TNMD and Tenascin-R of hBMSCs in the 3D-BMP-12 group were significantly higher than those in the material group(P<0.01),while the above indicators in the 3D-Triciribine group were significantly lower than those in the 3D-BMP-12 group(P<0.01).A significant increase of Akt and p-Akt expressions of hBMSCs in the 3D-BMP-12 group and a significant decrease in the 3D-Triciribine group were observed(P<0.01).Conclusion BMP-12 activates the Akt signaling pathway promoting the proliferation and tenogenic differentiation of hBMSCs in biomaterials.

关 键 词：骨形态发生蛋白-12 生物材料 间充质干细胞 细胞增殖 成肌腱分化 

分 类 号：R318.08[医药卫生—生物医学工程]