目的 探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)肝素化羟基磷灰石-磷酸三钙(HA-TCP)明胶支架的结构及理化性能,评估其对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSC)诱导分化作用。方法 将肝素结合到HA-TCP明胶支架上,检测支架的物理结构和机械性能;对不同肝素浓度的支架进行吸水率、VEGF负载、抑菌能力及骨髓间充质干细胞(BMSC)生长实验分析,来初步评价肝素化HA-TCP明胶支架的性能。结果 载VEGF肝素化HA-TCP明胶支架呈淡黄色,孔隙丰富,孔径范围100~400 μm,比表面积1.01±0.06 m2/g,平均介孔25.96 nm,弹性模量0.93 MPa,最大压缩强度2.40 MPa;三组HA-TCP明胶支架吸水率无明显差异,肝素化支架组比空白支架组有更好的VEGF结合、缓释能力(P<0.05),高浓度肝素组比低浓度肝素组能保持更久的VEGF活性浓度和抗菌能力(P<0.05)。BMSCs在支架上呈片状、岛状分布,肝素化支架组的细胞增殖活性明显强于空白组(P<0.05),产生更多的钙结节。结论 载VEGF肝素化HA-TCP明胶支架具有良好的理化性能;肝素化可提高支架的VEGF负载和释放能力、抑菌能力,促进BMSCs增殖和成骨分化,是一种有应用前景的骨组织工程材料。
Abstract
Objective To investigate the structure and physicochemical properties of vascular endothelial growth factor (VEGF) heparinized hydroxyapatite-tricalcium phosphate (HA-TCP) gelatin scaffold, and to evaluate its effect on the differentiation of bone marrow mesenchymal stem cells (BMSC).Methods Heparin was combined with HA-TCP gelatin scaffold to detect the physical structure and mechanical properties of the scaffold. To preliminarily evaluate the performance of heparinized HA-TCP gelatin scaffolds with different heparin concentrations, water absorption, VEGF loading, antibacterial ability and bone marrow mesenchymal stem cell (BMSC) growth were analyzed.Results The VEGF-loaded heparinized HA-TCP gelatin scaffold was pale yellow, with abundant pores (from 100 to 400μm), the specific surface area was 1.01±0.06 m2/g, the average mesoporous size was 25.96 nm, the elastic modulus was 0.93 MPa, and the maximum compressive strength was 2.40 MPa. There was no significant difference in the water absorption rate of HA-TCP gelatin scaffold among the three groups. The heparinized scaffold group had better VEGF binding and sustained release ability than the blank scaffold group (P<0.05), and the high heparin group could maintain the VEGF activity concentration and antibacterial ability longer than the low heparin group (P<0.05). BMSCs were distributed in sheets and islands on the scaffold. The cell proliferation activity of the heparinized scaffold group was significantly stronger than that of the blank group (P<0.05), and more calcium nodules were generated.Conclusions VEGF-loaded heparinized HA-TCP gelatin scaffold is characterized by good physical and chemical properties, and heparinization can improve the VEGF loading and releasing ability, bacteriostatic ability, and promote the proliferation and osteogenic differentiation of BMSCs, which is a promising bone tissue engineering material.
关键词
血管内皮生长因子 /
肝素 /
羟基磷灰石-磷酸三钙 /
组织工程
Key words
VEGF /
HA-TCP /
heparin /
tissue engineering
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