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  本文選題：再生絲素蛋白 + 肝素��； 參考：《東華大學》2012年碩士論文

【摘要】：滌綸材料具有良好的力學性能和化學穩(wěn)定性,目前已被廣泛應用于生物醫(yī)用材料領域,尤其是人造血管的構造。大中口徑的滌綸人造血管已經成功應用臨床數十年,創(chuàng)造了非常可觀的經濟利益和良好的社會利益。然而,滌綸小口徑人造血管的應用目前仍不理想。主要問題包括：短期內血栓形成和內皮化不良。因此,滌綸人造血管材料的改性一直是該領域的研究熱點之一。本研究擬采用等離子體和紫外輻照預改性滌綸人造血管材料并在其表面引入氨基,再將肝素和再生絲素蛋白分子吸附/接枝在滌綸改性材料表面,期望獲得力學性能優(yōu)良,生物學性能良好的滌綸血管材料,實現該人造血管移植早期具有良好的抗血栓性能,同時具有促進血管內皮細胞體內原位再生的良好性能。 本研究包括如下4項主要內容：一、采用常溫常壓等離子體和紫外光輻照預改性滌綸人造血管材料,優(yōu)化改性方法,并比較兩種改性方法對改性效果及改性材料力學性能的影響；二、以丙烯酰胺為“間隔臂”,通過Hoffman降解-共價縮合/正負離子鍵合-戊二醛交聯的方式制備再生絲素蛋白/肝素修飾滌綸人造血管改性材料,并對其進行表征；三、體外血液相容性評價。通過體外血栓形成實驗、溶血實驗、肝素緩釋等實驗評價再生絲素蛋白／肝素修飾滌綸人造血管改性材料的血液相容性；四、體外細胞相容性評價。通過體外細胞培養(yǎng)、MTT實驗及掃描電鏡觀察,考察再生絲素蛋白／肝素修飾滌綸人造血管改性材料的細胞相容性。 研究結果表明： 一、常溫常壓等離子體和紫外輻照均能將丙烯酰胺分子接枝到滌綸人造血管材料表面。然而,等離子體處理對滌綸材料力學性能有一定的損害,而紫外輻照影響不大。紫外輻照改性的優(yōu)化工藝為：二苯甲酮濃度0.2M/L、紫外光輻照功率5800μW/cm2、輻照時間20min、丙烯酰胺濃度2M/L。 二、如上改性的滌綸血管材料經Hoffman降解和再生絲素蛋白、肝素、戊二醛處理,可獲得再生絲素蛋白／肝素修飾滌綸人造血管改性材料。傅立葉紅外光譜表明兩物質相應的特征峰在滌綸材料表面得以表達,掃描電鏡觀察顯示再生絲素蛋白在滌綸表面分布較均勻,經高強度超聲波清洗60min后重量損失率不超過19%。 三、體外血栓形成實驗結果表明肝素修飾滌綸人造血管材料能有效地減少血栓形成,引入再生絲素蛋白能降低材料的溶血率。滌綸人造血管材料表面的肝素在PBS緩沖液中隨時間緩慢釋放,材料的抗凝血性能隨之逐漸衰減。溶血實驗結果表明,肝素修飾滌綸血管材料的溶血率低于5%,符合生物醫(yī)用材料應用標準。 四、體外細胞相容性實驗結果表明,再生絲素蛋白修飾滌綸人造血管改性材料具有良好的細胞相容性,再生絲素蛋白濃度的增加有利于血管內皮細胞的增殖。掃描電鏡觀察結果表明,血管內皮細胞能在改性材料表面良好黏附,活性較高。 本研究通過等離子體和紫外輻照的方法對滌綸人造血管材料表面進行了改性,并用再生絲素蛋白和肝素對材料表面進行了進一步修飾,從而獲得了力學性能及生物學性能良好的滌綸人造血管材料。本研究成果可為開發(fā)具有更好抗血栓性能的滌綸小口徑人造血管提供理論參考及技術支持。
[Abstract]:Polyester material has good mechanical property and chemical stability , and has been widely used in biomedical materials field , especially artificial blood vessel .

The research includes the following four main contents : Firstly , the modified method is optimized by using normal temperature and normal pressure plasma and ultraviolet radiation pre - modified polyester artificial blood vessel material , and the effect of two modification methods on the modification effect and the mechanical properties of modified materials are compared ;
secondly , preparing a regenerated silk fibroin / heparin modified polyester artificial blood vessel modifying material by using acrylamide as a " spacer arm " , and performing degradation - covalent condensation / positive - negative ionic bonding - glutaraldehyde cross - linking in a manner to characterize the modified polyester artificial blood vessel modified material ;
3 . In vitro blood compatibility evaluation . The blood compatibility of regenerated silk fibroin / heparin modified polyester artificial blood vessel modified material was evaluated by means of in vitro thrombosis experiment , hemolysis experiment , heparin slow release and so on .
In vitro cell culture , MTT assay and scanning electron microscope ( SEM ) were used to investigate the cellular compatibility of regenerated silk fibroin / heparin modified polyester artificial blood vessel modified material .

The results show that :

One , normal temperature and normal pressure plasma and ultraviolet irradiation can graft acrylamide molecules onto the surface of polyester artificial blood vessel material . However , the plasma treatment has certain damage to the mechanical properties of the polyester material , and the influence of ultraviolet radiation is not large . The optimization process of UV irradiation modification is as follows : benzophenone concentration 0.2M / L , UV irradiation power of 5500渭W / cm2 , irradiation time 20min , acrylamide concentration 2M / L .

secondly , the modified polyester vascular material can be treated with hosemide and regenerated silk fibroin , heparin and glutaraldehyde to obtain the regenerated silk fibroin / heparin modified polyester artificial blood vessel modified material , and the Fourier infrared spectrum shows that the corresponding characteristic peaks of the two substances are expressed on the surface of the terylene material , and the scanning electron microscope observation shows that the regenerated silk fibroin is distributed uniformly on the surface of the terylene material , and the weight loss rate after the high - intensity ultrasonic cleaning is not more than 19 percent after the high - intensity ultrasonic cleaning .

3 . The results of in vitro thrombosis show that heparin modified polyester artificial blood vessel material can effectively reduce thrombosis , and the introduction of regenerated silk fibroin can reduce the hemolysis rate of the material . The heparin in the surface of the polyester artificial blood vessel material is slowly released over time in PBS buffer solution , and the anti - coagulation property of the material is gradually attenuated . The hemolysis test results show that the hemolysis rate of the heparin - modified polyester blood vessel material is lower than 5 % , and conforms to the application standard of biomedical materials .

4 . The results of in vitro cell compatibility show that the regenerated silk fibroin modified polyester artificial blood vessel modified material has good cell compatibility , and the increase of the concentration of regenerated silk fibroin is beneficial to the proliferation of vascular endothelial cells . Scanning electron microscopy ( SEM ) shows that the vascular endothelial cells can adhere well on the surface of the modified material and have high activity .

The surface of polyester artificial blood vessel was modified by plasma and ultraviolet irradiation , and the surface of the material was further modified with regenerated silk fibroin and heparin , thus obtaining the polyester artificial blood vessel material with good mechanical properties and biological properties .
【學位授予單位】：東華大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.08

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本文編號：2069255