复合神经营养因子-3的静电纺丝高分子材料促进兔骨髓间充质干细胞增殖与分化

Electrospinning of Compound Neurotrophic Factor-3 Promotes Proliferation and Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells

目的探讨以静电纺丝技术制备的聚乳酸-羟基乙酸共聚物/-磷酸三钙(PLGA/-TCP)高分子材料与兔骨髓间充质干细胞(r BMSC)的生物相容性,并研究r BMSC在复合神经营养因子-3(NT-3)的PLGA/-TCP支架材料上的增殖与分化情况。方法选取齐齐哈尔医学院解剖学研究室于2016年7月购买的兔骨髓间充质干细胞。在正常条件下,单独培养的r BMSC为对照组;将r BMSC种植在PLGA/-TCP支架材料上为PT组;以100 ng/m L重组NT-3蛋白刺激的PT组为NT-3/PT组,各组均行成骨诱导实验21 d。利用CCK-8实验于成骨诱导第2、7、14、21天检测各组r BMSC增殖情况;ELISA实验检测各组r BMSC的碱性磷酸酶(ALP)、骨形态发生蛋白-1(BMP-1)及核心结合因子a-1(Cbfa-1)等蛋白含量。结果相对于PT组,NT-3/PT组的细胞增殖OD值在2 d(0.392±0.027)、7 d(0.594±0.024)、14 d(0.978±0.089)、21 d(1.124±0.045)均明显增高,差异有统计学意义(P<0.01);NT-3/PT组的ALP、BMP-1及Cbfa-1等蛋白含量[(1.525±0.352),(1.678±0.102),(1.104±0.076)pg/m L]均高于PT组[(1.154±0.741),(1.321±0.131),(0.852±0.067)pg/m L],差异有统计学意义(P<0.01)。结论复合NT-3的PLGA/-TCP静电纺丝高分子材料有效促进MSC增殖和向成骨细胞分化。

Objective This paper tries to investigate the biocompatibility of poly(DL-lactic-co-glycolic acid)/tricalcium phosphate(TCP-TCP)prepared by electrospinning with rabbit bone marrow mesenchymal stem cells(rBMSC)Proliferation and Differentiation on PLGA/-TCP Scaffolds with Nerve Trophoblast-3(NT-3).Methods Rabbit bone marrow mesenchymal stem cells purchased by anatomical research laboratory of Qiqihar medical college in July 2016 were selected.Under the normal situation,the rBMSC cultured alone as control group;the rBMSC implanted into the PLGA/-TCP polymeric material as PT group;the PT group stimulated by 100 ng/mL NT-3 recombination protein as NT-3/PT group,each group was subjected to osteogenic induction for 21 days.The CCK-8 assay was used to detect the proliferation of rBMSC on 2,7,14 and 21 d of osteogenic induction experiment in each group;ELISA experiment was used to detect the contents of alkaline phosphatase(ALP),bone morphogenetic protein-1(BMP-1),and core binding factor a 1(Cbfa-1)in each group.Results Compared with PT group,the cell proliferation OD values of NT-3/PT group were significantly elevated in 2 d(0.392±0.027),7 d(0.594±0.024),14 d(0.978±0.089),21 d(1.124±0.045),the difference was statistically significant(P<0.01);the contents of ALP,BMP-1 and Cbfa-1 protein in NT-3/PT group[(1.525±0.352),(1.678±0.102;),(1.104±0.076)pg/mL]were higher than those in PT group[(1.154±0.741),(1.321±0.131),(0.852±0.067)pg/mL],and the difference was statistically significant(P<0.01).Conclusion The electrospun polymeric material combined with neurotrophin-3 effectively promote rBMSC proliferation and osteogenic differentiation.