静电共纺聚合物聚己内酯-碳酸亚乙酯和血管内皮生长因子共混纳米纤维支架的体外细胞相容性研究  

Cellular compatibility of electrospun copolymer nanofibrous scaffolds composed of ethylenecarbonate-ε-caprolactone and vascular endothelial growth factor

作　　者：张婉[1] 张祥满[2] 史振宇[2] 虞奋 

机构地区：[1]复旦大学附属华东医院血管外科 [2]复旦大学附属中山医院血管外科

出　　处：《中华细胞与干细胞杂志（电子版）》2014年第3期10-14,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金面上项目(81370424)

摘　　要：目的通过静电纺丝技术将聚合物聚己内酯-碳酸亚乙酯[Poly(EC-CL)]和血管内皮生长因子(VEGF)为原料构建具有良好细胞相容性和机械性能的纳米支架。方法将VEGF与Poly(EC-CL)混合,配置成质量比为0 ng/g,10 ng/g,100 ng/g,1 μg/g的溶液,电压15 kV,流速2 ml/h电纺纳米纤维支架。扫描电镜检测电纺膜纳米形态,体外培养人脐静脉内皮细胞,细胞增殖试验及Live/Dead细胞试验检测电纺膜的生物相容性。各组电纺纤维直径比较以及细胞增殖MTS试验的(A)值比较均采用t检验。结果混合VEGF的poly(EC-CL)与纯poly(EC-CL)聚合物膜片相比,纤维形态没有分别,纳米纤维直径在(440 ± 55) nm,本研究增加VEGF含量,纤维直径没有明显变化(P> 0.05)。细胞增殖试验显示到培养第5 天时,细胞增殖加快,组织培养板和VEGF/poly(EC-CL)(100 ng/g、1μg/g)组明显高于poly(EC-CL)和VEGF/poly(EC-CL)(10 ng/g)(P < 0.05)。Live/Dead细胞试验显示四组膜片上,细胞均呈绿色荧光(活细胞),而且VEGF/ poly(EC-CL)(100 ng/g、1μg/g)膜片上细胞明显高于poly(EC-CL)和VEGF/ poly(EC-CL)(10 ng/g)两组。结论电纺技术构建的Poly(EC-CL)/VEGF共聚物具有良好的血管内皮细胞相容性,能够作为血管组织工程的基质材料。Objective To prepare a novel degradable nanofibrous scaffolds with copolymer ethylenecarbonate-^-caprolactone （Poly（EC-CL）） and vascular endothelial growth factor by electrospinning, and to test the biological compatibility and mechanical properties of the blended scaffolds. Methods Vascular endothelial growth factor （VEGF）was dissolved in 500 ul of M199 medium, and Poly（EC-CL） was dissolved in 1,1,1,2,2,2,-hexafluoro-2- propanol（HFIP） at a concentration of 10 % （w/v） for electrospinning. We mixed both solutions at a ratio of 1 μg/g, 100 ng/g, 10 ng/g, 0 ng/g （VEGF/poly（EC-CL）, wt%/wt%） respectively. The electrospinning process was performed at 15kV voltage with 15 cm distance between the needle tip and a rotating mandrel. The flow rate was 2mL/h. Scanning electron microscopy （SEM） was used to measure the fiber diameter distribution. The composition of fibrous membranes was analyzed by immunofluorescence. The cell proliferation and the biological compatibility were test by Live/Dead cells kit assay. Results The average diameter of these scaffolds was estimated to be （440 ± 55）nm. We increased the percentage ofVEGF/poly（EC-CL） from 10 ng/g to 1 μg/ g（w % ：w %）. The diameters of the scaffolds were not changed with the increase of VEGF, and there were no significant difference between the pure poly（EC-CL） and blended VEGF/poly（EC- CL）（P 〉 0.05）. Analysis of cell proliferation and Live/Dead cells confirmed that Poly （EC- CL） / VEGF films possessed excellent cell compatibility. Conclusion Poly （EC-CL） / VEGF copolymer has good cell compatibility with the endothelial cells. It can be used as scaffold for vascular engineering.

关 键 词：内皮细胞 血管内皮生长因子 纳米技术 相容性 

分 类 号：R318.08[医药卫生—生物医学工程]