蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验_《中国修复重建外科杂志》

作者：

赵亮 , 徐艳丽 , 邱慧 , 李敏 , 陈雨晴

福建师范大学生命科学学院（福州，350108）;

关键词：

组织工程血管蛛丝蛋白双层小直径血管支架静电纺丝血液相容性

DOI：

10.7507/1002-1892.20130176

视频：

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摘要 全文 图表 视频 参考文献 施引文献 补充材料

目的应用静电纺丝法制备一种蛛丝蛋白双层小直径血管支架，观察其体外血液相容性。方法将精氨酸-甘氨酸-天冬氨酸-重组蛛丝蛋白（pNSR16）、聚己内酯（polycaprolactone，PCL）、明胶（gelatin，Gt）、肝素（heparin，Hep）共混，以pNSR16∶PCL∶Hep质量比（W/W）为5∶85∶10的混合电纺溶液作为内层纺丝液，pNSR16∶PCL∶Gt质量比（W/W）为5∶85∶10的混合电纺溶液作为外层纺丝液，利用静电纺丝技术制备双层小直径血管支架（实验组）；以pNSR16∶PCL质量比（W/W）为5∶85的混合电纺溶液作为内层纺丝液制备支架作为对照组。扫描电镜观察实验组支架结构，比较两组溶血率、复钙化凝血时间、动态凝血时间以及体外血小板黏附、血小板激活实验结果。结果扫描电镜观察示实验组支架双层纤维不同，内层纤维直径分布较为均匀，孔隙相对较小；外层纤维直径差异较大，孔隙较大。实验组支架水接触角、溶血率、复钙化凝血时间和P-选择素表达量分别为（35 ± 3）°、1.2% ± 0.1%、（340 ± 11）s和0.412 ± 0.027，对照组分别为（70 ± 4）°、1.9% ± 0.1%、（260 ± 16）s和0.678 ± 0.031，各指标两组间比较差异均有统计学意义（P lt; 0.05）。实验组凝血时间曲线向下倾斜缓慢，与对照组相比前30 min内各时间点凝血指数均较高，比较差异有统计学意义（P lt; 0.05）。体外血小板黏附实验示实验组支架表面几乎无血小板黏附。结论静电纺丝技术可制备蛛丝蛋白双层小直径血管支架，且体外血液相容性好。

引用本文： 赵亮,徐艳丽,邱慧,李敏,陈雨晴. 蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验. 中国修复重建外科杂志, 2013, 27(7): 800-804. doi: 10.7507/1002-1892.20130176 复制

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14.	Kong X, Han B, Li H, et al. New biodegradable small-diameter artificial vascular prosthesis: A feasibility study. J Biomed Mater Res A, 2012, 100(6): 1494-1504.
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22.	Huang B, Chen YT, Tong FY, et al. Modification of hydrophilic property of polypropylene fiber film with water-soluble chitosan for improvement of blood compatibility. Advanced Science Letters, 2012, 7: 16-20.
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24.	国家质量监督检验检疫总局. GB/T 16886.4-2003/ISO 10993-4: 2002, 医疗器械生物学评价第4部分: 与血液相互作用试验选择. 北京: 中国标准出版社, 2003.
25.	周素琼, 李国明. 聚氨酯/硫酸酯化壳聚糖复合膜的制备及其血液相容性的研究. 化工时刊, 2007, 21(5): 1-4.
26.	Wang ZX, Sun B, Zhang M, et al. Functionalization of electrospun poly (ε-caprolactone) scaffold with heparin and vascular endothelial growth factors for potential application as vascular grafts. J Bioactive Compatible Polymer, 2013, 28(2): 154-166.
27.	Tan J, McClung WG, Brash JL. Non-fouling biomaterials based on blends of polyethylene oxide copolymers and polyurethane: simultaneous measurement of platelet adhesion and fibrinogen adsorption from flowing whole blood. J Biomater Sci Polym Ed, 2013, 24(4): 497-506.

1. Patterson JT, Gilliland T, Maxfield MW, et al. Tissue-engineered vascular grafts for use in the treatment of congenital heart disease: from the bench to the clinic and back again. Regen Med, 2012, 7(3): 409-419.
2. Hong Y, Ye SH, Nieponice A. A small diameter, fibrous vascular conduit generated from a poly (ester urethane) urea and phospholipid polymer blend. Biomaterials, 2009, 30(13): 2457-2467.
3. Phipps MC, Clem WC, Grunda JM, et al. Increasing the pore sizes of bone-mimetic electrospun scaffolds comprised of polycaprolactone, collagen I and hydroxyapatite to enhance cell infiltration. Biomaterials, 2012, 33(2): 524-534.
4. 虞希高, 陈正坚, 蒋宏亮, 等. 静电纺丝小直径人造血管机械力学性质. 中国生物医学工程学报, 2010, 29(5): 770-776.
5. Osnat H, David PK, Fritz V, et al. Spider and mulberry silkworm silks as compatible biomaterials. Composites Part B: Engineering, 2007, 38(3): 324-337.
6. Neppalli R, Marega, Marigo A, et al. Poly (epsilon-caprolactone) filled with electrospun nylon fibres: a model for a facile composite fabrication. Eur Polym J, 2010, 46(5): 968-976.
7. Kanda N, Morimoto N, Ayvazyan AA, et al. Evaluation of a novel collagen-gelatin scaffold for achieving the sustained release of basic fibroblast growth factor in a diabetic mouse model. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
8. Liu Y, Chen JR, Yang Y, et al. Improved blood compatibility of poly (ethylene terephthalate) films modified with L-arginine. J Biomater Sci Polym Ed, 2008, 19(4): 497-507.
9. 李敏, 陈登龙, 吴钦缘. 静电纺丝仪: 中国, 100457985[P]. 2007-02-07.
10. Xiang P, Li M, Zhang CY, et al. Cytocompatibility of electrospun nanofiber tubuLar scaffolds for small diameter tissue engineering blood vessels. Int J Biological Macromol, 2011, 49(3): 281-288.
11. 黄莹莹, 齐民, 杨大智, 等. 冠脉支架表面 PLGA 涂层制备及其血液相容性研究. 功能材料, 2006, 37(3): 411-414.
12. Li ZF, Ruckenstein E. Grafting of poly (ethylene oxide) to the surface of polyaniline films through a chlorosulfonation method and the biocompatibility of the modified films. J Colloid Interface Sci, 2004, 269(1): 62-71.
13. Wang HY, Feng YK, Zhao H, et al. Electrospun hemocompatible PU/Gelatin-Heparin nanofibrous bilayer scaffolds as potential artificial blood vessels. Macromolecular Research, 2012, 20(4): 347-350.
14. Kong X, Han B, Li H, et al. New biodegradable small-diameter artificial vascular prosthesis: A feasibility study. J Biomed Mater Res A, 2012, 100(6): 1494-1504.
15. Ayoub NA, Garb JE, Kuelbs A, et al. Ancient properties of spider silks revealed by the complete gene sequence of the prey-wrapping silk protein (AcSp1). Mol Biol Evol, 2013, 30(3): 589-601.
16. Ruckh TT, Carroll DA, Weaver JR, et al. Mineralization content alters osteogenic responses of bone marrow stromal cells on hydroxyapatite/polycaprolactone composite nanofiber scaffolds. J Funct Biomater, 2012, 3(4): 776-798.
17. Pok S, Myers JD, Madihally SV, et al. A multilayered scaffold of a chitosan and gelatin hydrogel supported by a PCL core for cardiac tissue engineering. Acta Biomater, 2013, 9(3): 5630-5642.
18. Harris NC, Davydova N, Roufail S, et al. The propeptides of VEGF-D determine heparin binding, receptor heterodimerization and effects on tumor biology. J Biol Chem, 2013, 288(12): 8176-8186.
19. Zhang H, Jia XL, Han F, et al. Dual-delivery of VEGF and PDGF by double-layered electrospunmembranes for blood vessel regeneration. Biomaterials, 2013, 34(9): 2202-2212.
20. Nowak E, Combes G, Stitt EH, et al. A comparison of contact angle measurement techniques applied to highly porous catalyst supports. Powder Technology, 2013, 233: 52-64.
21. 陈佳龙, 李全利, 陈俊英, 等. 利用胶原-肝素自组装多层膜改善纯钛表面血液相容性的研究. 功能材料, 2008, 39(8): 1363-1366.
22. Huang B, Chen YT, Tong FY, et al. Modification of hydrophilic property of polypropylene fiber film with water-soluble chitosan for improvement of blood compatibility. Advanced Science Letters, 2012, 7: 16-20.
23. Huang ZW, Ding TT, Sun J. Study of effect on cell proliferation and hemolysis of HAP and TCP nanometer particles. Advanced Materials Research, 2011, 378-379: 711-714.
24. 国家质量监督检验检疫总局. GB/T 16886.4-2003/ISO 10993-4: 2002, 医疗器械生物学评价第4部分: 与血液相互作用试验选择. 北京: 中国标准出版社, 2003.
25. 周素琼, 李国明. 聚氨酯/硫酸酯化壳聚糖复合膜的制备及其血液相容性的研究. 化工时刊, 2007, 21(5): 1-4.
26. Wang ZX, Sun B, Zhang M, et al. Functionalization of electrospun poly (ε-caprolactone) scaffold with heparin and vascular endothelial growth factors for potential application as vascular grafts. J Bioactive Compatible Polymer, 2013, 28(2): 154-166.
27. Tan J, McClung WG, Brash JL. Non-fouling biomaterials based on blends of polyethylene oxide copolymers and polyurethane: simultaneous measurement of platelet adhesion and fibrinogen adsorption from flowing whole blood. J Biomater Sci Polym Ed, 2013, 24(4): 497-506.

《中国修复重建外科杂志》

蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验

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《中国修复重建外科杂志》

蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验

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