BACKGROUND: Using bone tissue engineering methods for reconstruction of bone repair is an ideal treatment for bone defects, and it is crucial to combine biological scaffolds carrying growth factors with seed cells. OBJECTIVE: To observe the biocompatibility of rat bone marrow stromal stem cells (BMSCs) seeded onto vascular endothelial growth factor (VEGF)/bone morphogenetic protein 2 (BMP-2)-poly(lactic-co-glycolic acid) (PLGA) copolymer/ gelatin nanofibrous scaffolds.METHODS: Rat BMSCs were divided into five groups: blank scaffold group, BMP-2 group, VEGF group, VEGF/BMP-2 groups (1.2:1, 0.8:1, 0.4:1). In each group, the nanofibrous scaffold was placed at the bottom of the cell culture plate, and then rat BMSCs were seeded into the culture plate. Cell adhesion was observed under scanning electron microscope; cell proliferation was detected by cell counting kit-8; and ALP, RUNX-2 and OCN expression was determined by RT-PCR. RESULTS AND CONCLUSION: Under the scanning electron microscope, the BMSCs adhered to the scaffold and further grew into the scaffold. The adherent cells grew best in the 0.4:1 group, in which the cells were tightly integrated with the scaffold to form a cell-scaffold complex. Compared with the blank scaffold group, the cell proliferation and expression of ALP, RUNX-2 and OCN were both higher in the other groups, which was highest in the 0.4:1 group. To conclude, the VEGF/BMP-2-loaded PLGA copolymer/gelatin nanofibrous scaffold can promote cell adhesion, proliferation, osteogenic differentiation of rat BMSCs because of a good cytocompatibility. Moreover, the concentration ratio of VEGF/BMP-2 is optimized at 0.4:1.%背景:应用骨组织工程学方法进行骨修复重建是治疗骨缺损的理想方法,负载生长因子的生物支架与种子细胞的结合是关键.目的:将大鼠骨髓基质干细胞种植于负载血管内皮生长因子(vascular endothelial growth factor,VEGF)/骨形态发生蛋白2(bone morphogenetic protein-2,BMP-2)的聚乳酸-羟基乙酸共聚物(poly(lactic-co-glycolic acid),PLGA)/明胶纳米纤维支架,观察二者的生物相容性.方法:实验分为5组:空白支架组、BMP-2组、VEGF组、VEGF/BMP-2组(1.2:1)、VEGF/BMP-2组(0.8:1)、VEGF/BMP-2组(0.4:1),将各组纳米纤维支架放入细胞培养孔板底部,然后将大鼠骨髓基质干细胞接种于培养孔内,扫描电镜观察细胞在各组支架上的黏附情况,CCK-8法检测细胞增殖能力,RT-PCR检测成骨分化基因RUNX-2,ALP,OCN的表达.结果与结论:①在扫描电镜下可见细胞与支架黏附,并进入支架内部,VEGF/BMP-2(0.4︰1)组的细胞黏附生长状态最好,细胞与支架紧密结合成为支架细胞复合体;②负载生长因子的纳米纤维支架上的细胞增殖及成骨分化标记物ALP、RUNX-2和OCN表达量高于空白支架组,并且VEGF/BMP-2浓度比例为0.4︰1时成骨分化标记物表达量最高;③结果表明,负载VEGF/BMP-2的PLGA/明胶纳米纤维支架可促进大鼠骨髓基质干细胞的黏附、增殖及成骨分化,具备良好的细胞相容性,且VEGF/BMP-2浓度比例0.4:1为最佳比例.
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