生物可降解材料聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物与人脐静脉内皮细胞的细胞相容性

目的：探讨聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物（PEG-PLA-PGL/RGD）与人脐静脉内皮细胞的细胞相容性，为构建支架表面理想可降解载体材料奠定基础。方法：将分离培养的稳态生长的人脐静脉内皮细胞接种于PEG-PLA-PGL/RGD膜片上培养作为实验组，对照组为未放PEG-PLA-PGL/RGD膜片培养的人脐静脉内皮细胞，相差显微镜下观察细胞生长情况及细胞 计数， 采用MTT法检测细胞增殖指数及免疫组织化学S-P法检测人脐静脉内皮细胞FⅧ 。结果：人脐静脉内皮细胞在PEG-PLA-PGL / RGD膜片上生长良好，相差显微镜下观察种植在PEG-PLA-PGL/ RGD膜片上的第3代人脐静脉内皮细胞4～6 h后即开始贴壁、伸展；3 d后细胞开始呈集落样生长，生长较快；5 d后细胞集落开始融合，呈现特征性的鹅卵石样形态；经多次传代（超过7代）后，细胞呈长梭形或多角形,形态改变，与对照组比较差异无显著性。1周内台盼蓝法对细胞计数显示，实验组细胞较对照组无明显降低，细胞增长速度无明显降低（P> 0.05）；MTT法对种植后1、3、5及7 d的细胞增殖指数检测，两组比较差异均无显著性（P>0.05）。结论：人脐静脉内皮细在PEG-PLA-PGL/RGD上生长良好，PEG-PLA -PGL/RGD与人脐静脉内皮细胞具有良好的细胞相容性，是支架表面理想的可降解载体材料。

Cytocompatibility of biodegradable PEG-PLA-PGL/RGD with human umbiliacal vein endothelial cells cultivated in vitro

Objective To explore the cytocompatibility of biodegradable PEGPLA-PGL/RGD pol(ethyleneglycol)-b-poly(l-lactide)-b-poly(l-glutamic acid)] with human umbilical vein endothelial cells(HUVECs) and provide a basis for the development of biodegradable material for vessel scaffold.Methods The HUVESCs cultivated in vitro were inoculated onto PEG-PLA-PGL/RGD membrane and their growth and cell count were observed by phase contrast microscopy.Immunohistochemistry was used to examine the existence of factor Ⅷ expressed by HUVESs.The proliferation of cells was observed by MTT method.Results The observation result of phase contrast microscopy showed that endothelial cells grew well in PEG-PLA-PGL/RGD membrane and showed no significant difference with control.Cells attached to the PEG-PLA-PGL/ RGD and stretched after grafting 4—6 h.After 3 d,cells began to grow in colony rapidly.After 5 d,colony began to fuse and seemed like stone,the cells were shuttle and polygon in shape.Counting of HUVECs in 1 week: growth rate of cells in experimental group didn't decrease significantly as compared with control(P>0.05).There were no significant differences of the proliferation index of cells by MTT method 1,3,5 and 7 d after inoculation between experimental and control groups(P>0.05).Conclusion Tissue engineering blood vessel with normal cytocompatibility to a certain degree can be constructed through seeding cultured HUVECs on PEG-PLA-PGL/RGD scaffold and PEG-PLA-PGL/RGD is a perfect biodegradable carrier material.