家蠶再生絲素蛋白管狀支架的制備及性能研究
發(fā)布時間：2018-11-22 20:37
【摘要】：小口徑人造血管近年來一直是研究的重要課題,如何解決小口徑人造血管力學(xué)性能不足及凝血問題是關(guān)注的焦點(diǎn)。本文采用冷凍、風(fēng)干方法,選擇具有良好生物相容性的家蠶再生絲素蛋白(以下簡稱絲素蛋白)與作為交聯(lián)劑的聚乙二醇雙環(huán)氧丙烷醚(PEG-DE)復(fù)合作為內(nèi)膜和外膜、以蠶絲編織物作為中膜、模擬天然血管組織結(jié)構(gòu)構(gòu)建直徑小于6mm的絲素蛋白管狀支架。詳細(xì)探索了交聯(lián)比例(SF:PEG-DE,w/w)、SF濃度和冷凍溫度對管狀支架制備的影響,研究了管狀支架的形態(tài)結(jié)構(gòu),評價了PEG-DE作為交聯(lián)劑的絲素蛋白復(fù)合材料的細(xì)胞毒性。進(jìn)一步研究了管狀支架對SD大鼠成纖維細(xì)胞(L929)和人臍靜脈內(nèi)皮細(xì)胞(HUVECs)的生長支持作用。 以SF:PEG-DE=1:0.2～1:1.5制備的絲素蛋白管狀支架,隨著交聯(lián)比例中PEG-DE比重的增加,管狀支架成型更佳,管狀支架絲素蛋白結(jié)合率增加,當(dāng)SF:PEG-DE=1:1時管狀支架成型最好,絲素蛋白管狀支架中未交聯(lián)的絲素蛋白溶失最少,絲素蛋白分子充分發(fā)生了交聯(lián)反應(yīng),交聯(lián)程度達(dá)到最佳,支架內(nèi)表面微孔分布均勻。隨著絲素蛋白濃度由3%增加到6%,管狀支架成型更佳,絲素蛋白結(jié)合率上升,壁厚增加,孔徑變小。絲素蛋白濃度為3%時,管狀支架內(nèi)表面不能形成良好內(nèi)膜。隨著冷凍溫度(-20℃、-40℃、-80℃)的下降,管狀支架孔徑增大,孔壁因疏松使壁厚略有增加,對管狀支架中絲素蛋白的結(jié)合率沒有影響。 制備的絲素蛋白管狀支架松弛外徑約4.4～4.7mm,松弛內(nèi)徑為3.0～3.2mm,壁厚1mm,彎折直徑5.8～6.7mm,具有較好的彎曲性能。絲素蛋白管狀支架的軸向拉伸斷裂強(qiáng)度90MPa,軸向拉伸斷裂伸長率84%,徑向拉伸斷裂強(qiáng)度1000MPa,斷裂伸長率160%,同時具有良好的徑向壓縮回復(fù)率。結(jié)果表明,本文制備的管狀支架具有非常優(yōu)越的力學(xué)性能。另外,絲素蛋白管狀支架在37℃中的熱水溶失率1%。 MTT和細(xì)胞DNA總含量分析結(jié)果顯示,以PEG-DE作為交聯(lián)劑制備的絲素蛋白復(fù)合材料沒有明顯細(xì)胞毒性,L929細(xì)胞在共混膜上呈長梭形,鋪展緊密,細(xì)胞增殖活性強(qiáng)。采用熒光倒置顯微鏡、SEM、激光共聚焦顯微鏡觀察及MTT法測試結(jié)果顯示,絲素蛋白管狀支架能很好的支持L929細(xì)胞的粘附和增殖。進(jìn)一步研究了HUVECs在管狀支架內(nèi)表面的生長,同樣顯示了良好的粘附和增殖。
[Abstract]:In recent years, small diameter artificial blood vessel has been an important research topic, how to solve the problem of mechanical properties and coagulation of small diameter artificial blood vessel is the focus. In this paper, the regenerated silkworm silk fibroin with good biocompatibility and polyethylene glycol dioxide propane ether (PEG-DE) as crosslinking agent were selected as inner membrane and outer membrane by freezing and air drying methods. Silk fibroin tubular scaffolds with diameter less than 6mm were constructed by simulating the tissue structure of natural blood vessels. The effects of crosslinking ratio (SF:PEG-DE,w/w), SF concentration and freezing temperature) on the preparation of tubular scaffolds were investigated in detail. The cytotoxicity of fibroin composite with PEG-DE as crosslinking agent was evaluated. The effects of tubular scaffolds on the growth of SD rat fibroblasts (L929) and human umbilical vein endothelial cells (HUVECs) were further studied. With the increase of the proportion of PEG-DE in the crosslinking ratio of fibroin tubular scaffolds prepared by SF:PEG-DE=1:0.2~1:1.5, the tubular scaffolds were better formed, and the binding ratio of fibroin proteins in the tubular scaffolds increased. When SF:PEG-DE=1:1 was used, the tubular scaffold had the best shape, and the uncrosslinked fibroin in the fibroin tubular scaffold had the least dissolved loss, and the fibroin molecule had the best cross-linking reaction, the cross-linking degree was the best, and the distribution of micropores on the inner surface of the scaffold was uniform. With the increase of fibroin concentration from 3% to 6%, the tubular scaffold is better, the binding ratio of silk fibroin increases, the wall thickness increases, and the pore size becomes smaller. When fibroin concentration was 3, the inner surface of the tubular scaffold could not form a good intima. With the decrease of freezing temperature (-20 鈩，
本文編號：2350522
[Abstract]:In recent years, small diameter artificial blood vessel has been an important research topic, how to solve the problem of mechanical properties and coagulation of small diameter artificial blood vessel is the focus. In this paper, the regenerated silkworm silk fibroin with good biocompatibility and polyethylene glycol dioxide propane ether (PEG-DE) as crosslinking agent were selected as inner membrane and outer membrane by freezing and air drying methods. Silk fibroin tubular scaffolds with diameter less than 6mm were constructed by simulating the tissue structure of natural blood vessels. The effects of crosslinking ratio (SF:PEG-DE,w/w), SF concentration and freezing temperature) on the preparation of tubular scaffolds were investigated in detail. The cytotoxicity of fibroin composite with PEG-DE as crosslinking agent was evaluated. The effects of tubular scaffolds on the growth of SD rat fibroblasts (L929) and human umbilical vein endothelial cells (HUVECs) were further studied. With the increase of the proportion of PEG-DE in the crosslinking ratio of fibroin tubular scaffolds prepared by SF:PEG-DE=1:0.2~1:1.5, the tubular scaffolds were better formed, and the binding ratio of fibroin proteins in the tubular scaffolds increased. When SF:PEG-DE=1:1 was used, the tubular scaffold had the best shape, and the uncrosslinked fibroin in the fibroin tubular scaffold had the least dissolved loss, and the fibroin molecule had the best cross-linking reaction, the cross-linking degree was the best, and the distribution of micropores on the inner surface of the scaffold was uniform. With the increase of fibroin concentration from 3% to 6%, the tubular scaffold is better, the binding ratio of silk fibroin increases, the wall thickness increases, and the pore size becomes smaller. When fibroin concentration was 3, the inner surface of the tubular scaffold could not form a good intima. With the decrease of freezing temperature (-20 鈩，
本文編號：2350522
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