改性聚乙二醇水凝胶封装生长因子和内皮祖细胞及其可控性缓释  

Sustained release of polyethylene glycol-modified hydrogel-packaged growth factors and endothelial progenitor cells

作　　者：韩艳久[1] 刘国辉[1] 欧阳柳[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院西区创伤外科,湖北省武汉市430056

出　　处：《中国组织工程研究》2014年第30期4851-4856,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81141077)~~

摘　　要：背景:大量研究表明,为了实现聚合物材料的内皮细胞化,向材料表面负载生物活性因子是一种重要的手段,并且在聚合物材料表面引入人体血管内皮细胞将有助于提高材料的生物相容性。目的:合成封装碱性成纤维细胞生长因子、血管内皮生长因子和内皮祖细胞的水凝胶,观察生长因子的缓释效果,以及内皮祖细胞在水凝胶中的培养状态。方法:合成含精氨酸-甘氨酸-天冬氨酸肽段的聚乙二醇水凝胶,先后加入大鼠内皮祖细胞、碱性成纤维细胞生长因子、血管内皮生长因子溶液、基质金属蛋白酶反应底物肽段溶液。将上述水凝胶浸泡于PBS中,每12 h用ELISA法检测上清液中生长因子的水平;72 h后在水凝胶PBS溶液中再分别加入不同质量的基质金属蛋白酶2(100,1 000 ng)、基质金属蛋白酶9(100,1 000 ng),每12 h用ELISA法检测上清液中生长因子的水平。将封装内皮祖细胞的聚乙二醇水凝胶置于培养基中培养5 d,消化后用流式细胞仪检测存活细胞数。结果与结论:12-72 h内,碱性成纤维细胞生长因子、血管内皮生长因子的释放百分比一直维持在41%左右,72 h后分别加入不同质量的基质金属蛋白酶2、基质金属蛋白酶9,发现两种细胞因子的释放百分比呈稳步上升,72 h后已达95%以上,且基质金属蛋白酶质量越大两种因子的检测释放百分比越高。培养5 d后,在聚乙二醇水凝胶中仍有88.17%的内皮祖细胞存活。表明自组装聚乙二醇水凝胶既可以实现碱性成纤维细胞生长因子、血管内皮生长因子的可控性释放,又可以支持内皮祖细胞的增殖生长。BACKGROUND： Many studies have demonstrated that in order to achieve the endothelialization of the polymer materials, surface loading of bioactive factors is an important means and the introduction of human endothelial ceils on the surface of the polymer materials will contribute to increase the biocompatibility of materials. OBJECTIVE： To synthesize the polyethylene glycol hydrogel packaging vascular endothelial growth factor, basic fibroblast growth factor and endothelial progenitor cells, and to observe the sustained release of growth factors and culture status of endothelial progenitor cells. METHODS： The polyethylene glycol-modified hydrogel containing arginine-glycine-aspartic acid peptide was synthesized, and then, rat endothelial progenitor cells, basic fibroblast growth factor, vascular endothelial growth factor, matrix metalloproteinases were successively added. The hydrogel were immersed in PBS, and ELISA was used to test the levels of growth factors in supernatants every 12 hours. Matrix metalloproteinases-2 （100, 1 000 ng） and matrix metalloproteinases-9 （100, 1 000 ng） at different mass were added into the PBS containing hydrogel after 72 hours. ELISA was also used to test the levels of growth factors in supernatants every 12 hours. The hydrogel encapsulating endothelial progenitor cells was cultured in a culture medium for 5 days, and the number of survival cells was counted by a flow cytometer after digestion.RESULTS AND CONCLUSION： Within 12-72 hours, the release percentage of basic fibroblast growth factor and vascular endothelial growth factor remained at about 41%. After 72 hours, the release percentage of both growth factors was found to grow steadily when matrix metalloproteinases-2 （100, 1000 ng） and matrix metalloproteinases-9 （100, 1 000 ng） at different mass were added, which reached 95%. The release percentage was increased with the increasing mass of matrix metalloproteinases. After 5 days of culture, 88.17% cans still survived in the hydrogel. These findings ind

关 键 词：生物材料 缓释材料 聚乙二醇 生长因子 可控性释放 内皮祖细胞 细胞增殖 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]