Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3493
標題: Evaluation of endothelial cells-seeded polyurethane small-diameter vascular grafts
內皮細胞植覆於聚胺基甲酸酯人工小血管之評估
作者: 林大鈞
Lin, Ta-Chun
關鍵字: vascular graft
人工小血管
endothlial cell
polyurethane
pellethane 2363-80A
bioreactor
dynamic culture
內皮細胞
聚胺基甲酸酯
動態培養
生物反應器
動物實驗
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(PU)-Pellethane 2363-80A為材料,嘗試以不同的加工法,包括混鹽浸鑄法、非溶劑水凝法及混鹽浸鑄水凝法,製作成具微孔洞相連結構之小口徑人工血管;其中混鹽浸鑄水凝法可得到良好機械性質之人工小血管,經電子顯微鏡(SEM)發現其具極佳的微孔洞相連結構,且具有混合式孔隙度。 以混鹽浸鑄法製成人工小血管,並以明膠與含RGD的重組蛋白質CBD-RGD加以改質。以三段式重力法將人臍靜脈內皮細胞株植覆於血管內壁,再以靜態或動態環境下培養,培養一週後,發現細胞並無明顯的生長。進行無血清培養液沖刷實驗,計算細胞殘餘率,可評估其抗沖刷能力,並以SEM作為輔證。結果發現,靜態培養一週後,雖然細胞並無增生,但在沖刷後所得細胞殘餘率為98.9﹪,抗沖刷能力極佳;在1 rpm以上的動態培養一週後,內皮細胞數量減少為一半,在沖刷後細胞殘餘率約僅30%,而在1 rpm以下動態培養的情形有稍微改善,未來應繼續降低轉速,以其找出最佳培養環境。 經明膠改質的內徑3mm、長3∼4 cm人工小血管,在植入豬的頸動脈後,可利用都卜勒超音波觀察血管內部血液流動的情形。經過2個月後血管阻塞,取出人工血管接合處附近動脈血管作組織切片,發現有內膜增生的現象。 自豬體內取得的頸靜脈內皮細胞,體外培養後可達第3代,至第4代細胞會變形呈多觸角狀。經VWF鑑定1及2代仍有內皮細胞之特性。
A medical grade polyurethane, Pellethane 2363-80A, was used to fabricate the sponge type small-diameter vascular grafts in this study, using three different processes, including salt- impregnated, salt-impregnated water-coagulation and non-solvent coagulation. The grafts had an open-cell structure. Among them, those fabricated by salt-impregnated water-coagulation method had the best mechanical properties and composite porosity. The grafts fabricated by salt-impregnated method were modified with gelatin and CBD-RGD (celluose-binding domain RGD-containing protein). Human umbilical vein endothelial cell lines were seeded on the graft lumen by three-step gravitational method. After being seeded, the grafts were cultured in static or dynamic conditions for a week. The grafts were perfused in vitro for 3 hours and the cell retention was measured. In static conditions, the cell retention was 98.8%. In dynamic conditions, the cell retention was greatly reduced when the rotational speed for the culture deck was above 1 rpm. When the speed was below 1 rpm, the cell retention was better. The grafts modified with gelatin (3-4 cm in length and 3 mm in diameter) were implanted into porcine carotid arteries. The patency was evaluated by monitering the blood-flow using Doppler-ultrasound. After two months, the thombus was found. The grafts and the surrounding tissues were resected for histological studies. There was intimal hyperplasia observed near anastomotic site. The porcine jugular vein endothelial cells could be harvested and cultured up to the third passage. After that, the cells became elongated and lost the phenotypic morphology. The cells for the passages 1 and 2 contained VWF antigens on their surface.
URI: http://hdl.handle.net/11455/3493
Appears in Collections:化學工程學系所

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