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目的 研究在纳米纤维表面共培养人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)对人和大鼠骨髓间充质干细胞(human and rat bone marrow mesenchymal stem cells, hMSCs and rMSCs)向成骨/成血管分化的影响,为骨组织工程血管化及骨快速修复提供方法.方法 将聚乳酸-羟基乙酸共聚物[poly (lactic-co-glycolic acid), PLGA]静电纺丝纳米纤维进行氨气等离子体处理来接枝氨基基团,用扫描电子显微镜(scanning electronic microscopy, SEM)观察处理前后纤维形貌变化及直径分布;将HUVECs与hMSCs和rMSCs共培养(hMIX, rMIX)于修饰前后的PLGA纳米纤维表面,在培养1, 3 (4), 7 d后用细胞计数试剂盒(cell counting kit-8, CCK-8)检测细胞增殖,培养14 d后CD31免疫荧光染色检测成血管分化情况,培养21 d后茜素红S (alizarin red S,ARS)染色钙结节检测成骨分化情况.结果 SEM形貌观察结果显示经氨气等离子体处理后PLGA纳米纤维的直径及粗糙度略有增加;CCK-8结果显示氨基修饰后的纳米纤维表面更有利于rMIX的黏附和增殖,而对于hMIX的黏附和增殖影响很小;ARS染色结果显示氨基修饰的表面可以促进rMSCs成骨分化,而对hMSCs影响不大;CD31染色结果显示,HUVECs共培养及氨基修饰纳米纤维可以促进hMSCs成血管分化,而对rMSCs成血管分化没有促进作用.结论 氨基修饰的纳米纤维表面培养hMSCs-HUVECs可以有效促进组织工程骨血管化,在骨组织工程中有一定的应用前景.“,”To study the osteogenic and angiogenic effects of co-culture HUVECs and r/hMSCs on electrospun nanofibers and provide new strategy for angiogenesis and fast-repair of bone. Method PLGA nanofibers were treated with ammonia plasma to graft amine groups; The morphology of nanofibers before and after treatment was observed by SEM, and the distribution of diameter of nanofibers was measured by ImageJ; HUVECs were co-cultured with h/rMSCs (hMIX, rMIX) onto PLGA nanofibers and amine group grafted nanofibers; CCK-8 was used to test the attachment and proliferation of cells at day 1, 3 (4), 7 after seeded; CD31 immunofluorescence staining was employed to detect the angiogenic differentiation of MSCs after 14 days cultured; ARS staining was used to detect the calcium deposit to show osteogenesis condition. Results The SEM results showed that the diameter and roughness of nanofibers was increased after plasma treatment. CCK-8 results showed that amine group grafted nanofibers was much more beneficial to the attachment and proliferation of rMIX rather than hMIX. CD31 staining results indicated the promotion angiogenic effects of co-culturing and amine grafting to hMSCs. However, the enhancement effects were not observed on rMSCs. Conclusions HUVECs and hMSCs co-cultured onto PLGA NF-NH2 can enhance blood vessel formation in bone tissue engineering and has promising application potential.