蛛絲蛋白雙層小直徑血管支架的制備及其血液相容性體外實驗_《中國修復重建外科雜志》

作者：

趙亮 , 徐艷麗 , 邱慧 , 李敏 , 陳雨晴

福建師范大學生命科學學院（福州，350108）;

關鍵詞：

組織工程血管蛛絲蛋白雙層小直徑血管支架靜電紡絲血液相容性

DOI：

10.7507/1002-1892.20130176

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目的應用靜電紡絲法制備一種蛛絲蛋白雙層小直徑血管支架，觀察其體外血液相容性。方法將精氨酸-甘氨酸-天冬氨酸-重組蛛絲蛋白（pNSR16）、聚己內酯（polycaprolactone，PCL）、明膠（gelatin，Gt）、肝素（heparin，Hep）共混，以pNSR16∶PCL∶Hep質量比（W/W）為5∶85∶10的混合電紡溶液作為內層紡絲液，pNSR16∶PCL∶Gt質量比（W/W）為5∶85∶10的混合電紡溶液作為外層紡絲液，利用靜電紡絲技術制備雙層小直徑血管支架（實驗組）；以pNSR16∶PCL質量比（W/W）為5∶85的混合電紡溶液作為內層紡絲液制備支架作為對照組。掃描電鏡觀察實驗組支架結構，比較兩組溶血率、復鈣化凝血時間、動態凝血時間以及體外血小板黏附、血小板激活實驗結果。結果掃描電鏡觀察示實驗組支架雙層纖維不同，內層纖維直徑分布較為均勻，孔隙相對較小；外層纖維直徑差異較大，孔隙較大。實驗組支架水接觸角、溶血率、復鈣化凝血時間和P-選擇素表達量分別為（35 ± 3）°、1.2% ± 0.1%、（340 ± 11）s和0.412 ± 0.027，對照組分別為（70 ± 4）°、1.9% ± 0.1%、（260 ± 16）s和0.678 ± 0.031，各指標兩組間比較差異均有統計學意義（P lt; 0.05）。實驗組凝血時間曲線向下傾斜緩慢，與對照組相比前30 min內各時間點凝血指數均較高，比較差異有統計學意義（P lt; 0.05）。體外血小板黏附實驗示實驗組支架表面幾乎無血小板黏附。結論靜電紡絲技術可制備蛛絲蛋白雙層小直徑血管支架，且體外血液相容性好。

引用本文： 趙亮,徐艷麗,邱慧,李敏,陳雨晴. 蛛絲蛋白雙層小直徑血管支架的制備及其血液相容性體外實驗. 中國修復重建外科雜志, 2013, 27(7): 800-804. doi: 10.7507/1002-1892.20130176 復制

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, 陳正堅, 蔣宏亮, 等. 靜電紡絲小直徑人造血管機械力學性質. 中國生物醫學工程學報, 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, 陳登龍, 吳欽緣. 靜電紡絲儀: 中國, 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.	黃瑩瑩, 齊民, 楊大智, 等. 冠脈支架表面 PLGA 涂層制備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.	陳佳龍, 李全利, 陳俊英, 等. 利用膠原-肝素自組裝多層膜改善純鈦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.	國家質量監督檢驗檢疫總局. GB/T 16886.4-2003/ISO 10993-4: 2002, 醫療器械生物學評價第4部分: 與血液相互作用試驗選擇. 北京: 中國標準出版社, 2003.
25.	周素瓊, 李國明. 聚氨酯/硫酸酯化殼聚糖復合膜的制備及其血液相容性的研究. 化工時刊, 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, 陳正堅, 蔣宏亮, 等. 靜電紡絲小直徑人造血管機械力學性質. 中國生物醫學工程學報, 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, 陳登龍, 吳欽緣. 靜電紡絲儀: 中國, 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. 黃瑩瑩, 齊民, 楊大智, 等. 冠脈支架表面 PLGA 涂層制備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. 陳佳龍, 李全利, 陳俊英, 等. 利用膠原-肝素自組裝多層膜改善純鈦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. 國家質量監督檢驗檢疫總局. GB/T 16886.4-2003/ISO 10993-4: 2002, 醫療器械生物學評價第4部分: 與血液相互作用試驗選擇. 北京: 中國標準出版社, 2003.
25. 周素瓊, 李國明. 聚氨酯/硫酸酯化殼聚糖復合膜的制備及其血液相容性的研究. 化工時刊, 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.

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蛛絲蛋白雙層小直徑血管支架的制備及其血液相容性體外實驗

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《中國修復重建外科雜志》

蛛絲蛋白雙層小直徑血管支架的制備及其血液相容性體外實驗

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