RGD修饰的内皮祖细胞捕获支架涂层材料体外生物学性能研究

发布时间：2018-06-09 06:24

本文选题：内皮祖细胞捕获支架 + PEG-PLA-PGL/RGD涂层　；参考：《吉林大学》2014年硕士论文

【摘要】：研究背景冠状动脉支架置入术后支架内再狭窄和血栓形成是限制冠心病介入治疗发展的瓶颈，支架置入后表面早期内皮化从而完成内膜修复是解决再狭窄和血栓形成的关键。内皮祖细胞（endothelial progenitorcell,EPC）捕获支架通过捕获EPC加速内皮化，是目前研究的热点。前期本研究组针对第一代药物涂层支架延迟内皮化的问题，将具有捕获EPC功能的精氨酸-甘氨酸-天门冬氨酸三肽(Arg-Gly-Asp,RGD)接枝于可降解聚合物聚乙二醇-聚乳酸-聚谷氨酸共聚物[poly(ethyleneglycol)-poly (L-lactic acid)-poly (L-glutamate acid), PEG-PLA-PGL]涂层上，成功研制了新型EPC捕获支架即PEG-PLA-PGL/RGD涂层支架，并证明了其有良好的细胞相容性。本课题在前期研究基础上，进一步对PEG-PLA-PGL/RGD涂层支架其他生物学性能进行研究。研究目的评价PEG-PLA-PGL/RGD涂层支架的体外血液相容性；在证明了有良好生物相容性基础上探讨对内皮细胞粘附稳定性的影响，从而为合格新型EPC捕获支架研制提供更多的实验基础。研究方法以裸钢片、PEG-PLA-PGL聚合物涂层钢片为对照组，通过体外溶血实验、血小板吸附数量测定、抗凝血时间测定及蛋白吸附评价RGD修饰PEG-PLA-PGL共聚物（PEG-PLA-PGL/RGD）涂层钢片的体外血液相容性。以PEG-PLA-PGL膜片为对照组,PEG-PLA-PGL/RGD膜片为实验组，体外在2组材料表面种植人脐静脉内皮细胞,流体条件下观察在T型管及直型管不同位置、不同流速下RGD对材料表面细胞黏附稳定性的影响。研究结果体外血液相容性结果：裸钢片、 PEG-PLA-PGL组、PEG-PLA-PGL/RGD组三组钢片溶血率均＜5%，无溶血作用；与标本血比较，三组钢片APTT均表现出明显差异（P＜0.05），均能改善标本血抗凝血性能；PEG-PLA-PGL涂层与裸钢片相比，在各项所测指标中无统计学差异；接枝RGD后PEG-PLA-PGL/RGD涂层在血小板吸附及白蛋白吸附方面优于裸钢片，有统计学差异（P＜0.05），一定程度上提高了裸钢片的血液相容性。细胞粘附稳定性结果：T型管及直型管各对应位置，，PEG-PLA-PGL共聚物表面结合RGD后,细胞残余率明显增加（P＜0.05）；无论在对照组还是在实验组，T型分叉前A位置细胞残余率都要高于分叉处及分叉后的细胞残余率（P0.05）。研究结论体外实验证实PEG-PLA-PGL/RGD涂层支架材料具有良好的血液相容性。体外实验表明RGD可以提高PEG-PLA-PGL支架涂层材料的细胞粘附稳定性，包括在T型管分叉处；但细胞粘附稳定性受T型管分叉的影响。
[Abstract]:Background Intra-stent restenosis and thrombosis after coronary stent implantation are the bottleneck to restrict the development of coronary intervention. Early endothelialization of the surface after stent implantation is the key to resolve restenosis and thrombosis. Endothelial progenitor cells (EPC) capture scaffolds accelerate endothelialization by capturing EPC, which is a hot research topic at present. Earlier, this group focused on the delayed endothelialization of first-generation drug-coated stents. Arginine glycine aspartic acid tripeptide Arg-Gly-AspRGD was grafted onto poly(ethyleneglycol)-poly L-lactic acid)-poly L-glutamate acid (PEG-PLA-PGL) coating. A novel EPC capture scaffold PEG-PLA-PGL / RGD coated scaffold was successfully developed and proved to have good cytocompatibility. On the basis of previous studies, the other biological properties of PEG-PLA-PGL / RGD coated scaffolds were further studied in order to evaluate the in vitro blood compatibility of PEG-PLA-PGL / RGD coated scaffolds. On the basis of proved good biocompatibility, the effect on the adhesion stability of endothelial cells was discussed, which provided more experimental basis for the development of qualified new EPC capture scaffold. The study method was based on PEG-PLA-PGL polymer coated steel sheet as control group. The in vitro blood compatibility of PEG-PLA-PGL modified PEG-PLA-PGL coated steel sheet was evaluated by in vitro hemolysis test, platelet adsorption quantity measurement, anticoagulant time measurement and protein adsorption. The PEG-PLA-PGL / RGD membrane was used as the control group. Human umbilical vein endothelial cells were implanted on the surface of the two groups in vitro. The effect of RGD on the stability of cell adhesion on the surface of materials at different flow rates. Results the results of in vitro blood compatibility were as follows: the hemolysis rate of the steel sheets in the bare steel sheet and the PEG-PLA-PGL / RGD group was less than 5, and there was no hemolytic effect in the RGD group, and compared with the sample blood, the hemolysis rate of the RGD was lower than that of the PEG-PLA-PGL / RGD group. The APTT of the three groups showed significant difference (P < 0.05), which could improve the blood anticoagulant performance of the samples. The PEG-PLA-PGL coating had no statistical difference compared with the bare steel sheet. The PEG-PLA-PGL / RGD coating was better than bare steel in platelet adsorption and albumin adsorption after graft RGD (P < 0.05), which improved the blood compatibility of bare steel to some extent. The results of cell adhesion stability showed that the PEG-PLA-PGL copolymers were attached to RGD on the surface of the PEG-PLA-PGL copolymers. The residual rate of cells increased significantly (P < 0.05), both in the control group and in the experimental group before the T-type bifurcation was higher than that in the branched and branched cells. Conclusion the PEG-PLA-PGL / RGD coating scaffold was confirmed by in vitro experiments. [WT5 "HZ] [WT5BZ] the results show that the cell residual rate of the PEG-PLA-PGL / RGD coating scaffolds is higher than that of the control group and the experimental group. The material has good blood compatibility. In vitro experiments showed that RGD could improve the cell adhesion stability of PEG-PLA-PGL scaffold, including at the junction of T-tube, but the cell adhesion stability was affected by T-tube bifurcation.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R541.4;R318.08

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