经基因复合物改性的PCL-PIBMD静电纺丝人工血管支架的研究

发布时间：2019-02-28 15:20

【摘要】：目前,人工血管移植术是医治心血管疾病的一种有效的手术方法。然而,由于人工血管表面缺乏活性内皮层,人工血管特别是小口径人工血管经常会遇到通畅率低和再狭窄等问题,从而限制了其在临床上的应用。相关研究发现,人工血管的快速内皮化有助于改善上述问题。伴随着基因治疗的发展,使用基因转染技术加速血管内皮细胞增殖受到了越来越多的关注。本论文利用静电纺丝以及静电喷雾技术,成功构建了由靶向内皮细胞增殖的基因复合物修饰的电纺血管纤维膜,在此基础上,制备出了复合小口径人工血管,其研究结果概括如下:1.通过静电喷雾技术,将具有靶向性的聚阳离子基因载体微粒(MPs)以及其与pZNF580基因形成的复合物(MPs-pZNF580),负载在聚己内酯(PCL)和聚-3S-吗啉-1-异丁基-2,5-二酮(PIBMD)嵌段共聚物所制备的电纺PCL-PIBMD纤维膜表面,随着MPs以及MPs-pZNF580的加入,PCL-PIBMD纤维的直径和机械性能没有明显变化,但是改性后的PCL-PIBMD/MPs以及PCL-PIBMD/MPspZNF580纤维膜的亲水性能有所提高,通过血小板黏附实验证明,PCLPIBMD/MPs以及PCL-PIBMD/MPs-pZNF580纤维膜比空白PCL-PIBMD纤维膜具有更好的血液相容性。细胞活性测试以及荧光染色表明,经过7天的培养,PCLPIBMD/MPs-pZNF580纤维膜表面相对细胞数达到121.5%±1.2%,细胞密度达到749细胞/平方毫米,表明该纤维膜能够明显地促进内皮细胞(EA.hy926 cells)的增殖。2.采取大鼠背部皮下埋植模型评价纤维膜的组织相容性。相较于PCLPIBMD以及PCL-PIBMD/MPs对照组,由MPs-pZNF580修饰的PCL-PIBMD纤维膜表现出最少的急性炎症反应。3.利用多层静电纺丝技术,制备了以PCL-PIBMD/MPs-pZNF580为内层,聚碳酸酯-聚氨酯(PCU)为外层的复合小口径人工血管,兔颈动脉移植实验结果显示该复合小口径血管具有良好的通畅率,表明其在血管组织工程中具有良好的应用前景。
[Abstract]:At present, artificial vascular transplantation is an effective surgical method for the treatment of cardiovascular diseases. However, due to the lack of active endodermis on the surface of artificial vessels, especially small-diameter artificial vessels, often encounter problems such as low patency rate and restenosis, which limits its clinical application. Related studies have found that the rapid endothelium of artificial blood vessels can help to improve the above-mentioned problems. With the development of gene therapy, more and more attention has been paid to accelerating the proliferation of vascular endothelial cells by gene transfection. In this paper, the electrospun vascular fiber membrane was successfully constructed by electrospinning and electrostatic spray. On the basis of this, the composite small diameter artificial blood vessel was prepared, which was modified by the gene complex of endothelial cell proliferation. The results of the study are summarized as follows: 1. The targeting polycationic gene carrier particle (MPs) and its complex with pZNF580 gene (MPs-pZNF580) were prepared by electrostatic spray technique. Supported on the surface of electrospun PCL-PIBMD fiber membrane prepared by polycaprolactone (PCL) and poly-3S-morpholine-1-isobutyl-2,5-dione (PIBMD) block copolymer, with the addition of MPs and MPs-pZNF580, The diameter and mechanical properties of PCL-PIBMD fiber did not change obviously, but the hydrophilicity of modified PCL-PIBMD/MPs and PCL-PIBMD/MPspZNF580 fiber membrane was improved. PCLPIBMD/MPs and PCL-PIBMD/MPs-pZNF580 fiber membrane have better blood compatibility than blank PCL-PIBMD fiber membrane. Cell viability test and fluorescence staining showed that after 7 days of culture, the relative number of cells on the surface of PCLPIBMD/MPs-pZNF580 fiber membrane reached 121.5% 卤1.2%, and the cell density reached 749 cells / square millimeter. The results show that the fiber membrane can significantly promote the proliferation of endothelial cells (EA.hy926 cells). 2. The histocompatibility of fibrous membrane was evaluated by subcutaneous implantation model in rats. Compared with the PCLPIBMD and PCL-PIBMD/MPs control groups, the PCL-PIBMD fibrous membrane modified by MPs-pZNF580 showed the least acute inflammatory reaction. 3. The composite small diameter artificial blood vessel with PCL-PIBMD/MPs-pZNF580 as inner layer and polycarbonate-polyurethane (PCU) as outer layer was prepared by multi-layer electrospinning technology. The experimental results of rabbit carotid artery transplantation show that the composite small diameter vessel has a good patency rate, indicating that it has a good application prospect in vascular tissue engineering.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R318.08;TQ340.64

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