Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3535
標題: Endothelial cell seeding on small diameter vascular grafts with modified inner surfaces
小口徑人工血管經表面改質後植覆內皮細胞之研究
作者: Sung, Song-huan
孫崧桓
關鍵字: small diameter vascular graft
人工小血管
endothelial cell seeding
canine jugular vein endothelial cell
內皮細胞植覆
狗頸靜脈內皮細胞
出版社: 化學工程學系
摘要: 本研究使用醫用級的聚胺基甲酸酯(polyurethane,PU)-Pellethane 2363-80A,製作成小口徑人工血管的基材,經表面改質後植覆上內皮細胞。經實驗證實以此製作出的小血管具有良好的血液相容性,也有一良好的細胞覆蓋率,相信若應用於未來的動物移植實驗時,會有良好的暢通率。 具微孔洞結構的小血管是以混鹽浸鑄法(salt-impregnated)來製造,其孔洞小於37μm。經由調整PU的濃度以及混鹽比例的多寡,可以控制小血管的彈性模數(E')為0.53±0.08 MPa,進而達到小血管順應性的匹配。 在小血管表面上交聯上一層明膠(gelatin)後,其厚度約為40μm,並吸附上含RGD序列(arginine-glycine-aspartate,RGD motif)的蛋白質,此為我們最佳改質的小血管。 在血小板貼附測試上,利用富血小板血漿來進行測試(platelet rich plasma,PRP)。由平均血小板活化個數(normalized number of activated platelets)來看,經最佳改質的表面其平均活化個數只有未改質表面的不到一半,可見經由最佳改質的程序後,確實能提高血液相容性。 在前述的人工小血管植覆上內皮細胞株後,我們利用體外的沖刷(in vitro perfusion)來測試細胞在小血管表面上的貼附能力。在經由最佳改質的表面上,其細胞殘留率為未改質者之3倍,可見經由最佳改質的程序後,的確能增加細胞在小血管表面上的貼附能力。 由狗頸靜脈取得之內皮細胞,經體外培養之後,植覆在最佳改質的小血管表面上,經由染色觀察後發現依然有很高的覆蓋率,可見最佳改質的小血管表面對內皮細胞株與頸靜脈內皮細胞均具有良好的貼附性。
In this study, we used a medical grade polyurethane, Pellethane 2363-80A, to fabricate sponge type small-diameter vascular grafts by a salt casting technique. The grafts were compliance-matched (with a storage modulus was 0.53±0.08 MPa) by a salt casting technique. The inner surface of grafts was coated with a thin layer (~ 40 μm) of gelatin crosslinked by epoxide. Then a special RGD-containing peptide was coated on the gelatin layer. The platelet adhesion and activation was evaluated by using platelet rich plasma (PRP). It was observed that the normalized number of activated platelets on the best modified surface (i.e. that modified by both gelatin and RGD) was only about a half of that on the surface without modification. Therefore, surface modification by gelatin and RGD indeed increased the blood compatibility. After endothelial cells were seeded onto the inner surface of the vascular grafts, the grafts were perfused in vitro and the cell retention was measured. The cell number on the surface modified by gelatin and RGD after in vitro perfusion was enhanced by a factor of three. The primary endothelial cells were isolated from canine external jugular vein, cultured and seeded onto the vascular graft. The cell coverage was observer by staining. The primary cells also had a high cell coverage on the surface modified by gelatin and RGD. We believed that these experimental results suggested patency of such grafts when implanted in the animal model in the future.
URI: http://hdl.handle.net/11455/3535
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