人脐静脉内皮细胞与表面改性的PHBHHx膜的体外相容性

Compatibility of human umbilical vein endothelial cells and surface-modified PHBHHx film in vitro

目的评价人脐静脉内皮细胞(HUVECs)在含有不同组分中药涂层、碱处理前后的3-羟基丁酸和3-羟基已酸共聚酯(PHBHHx)膜上的生长状况,为研究PHBHHx膜与内皮细胞的生物相容性提供实验依据。方法用Baudine改良法分离培养人脐静脉内皮细胞,并用免疫组织化学方法鉴定;将已传至第3代人脐静脉内皮细胞接种于材料表面,培养8、12、24h,用扫描电镜观察人脐静脉内皮细胞在不同表面上黏附形态的变化,同时用细胞免疫荧光标记法比较其在不同膜上的分布情况,最后用四甲基偶氮唑盐(MTT)法检测各组细胞的活力。结果成功分离出人脐静脉内皮细胞,Ⅷ因子及血管内皮细胞受体Ⅱ(FLK-1)染色阳性;人脐静脉内皮细胞培养结果表明,细胞在各组材料表面均呈良好生长,并大量增殖。MTT分析结果显示,碱处理的PHBHHx膜及添加5%、10%中药涂层的材料对人脐静脉内皮细胞的体外生长、增殖有促进作用;扫描电镜和荧光显微镜下可见,碱处理的PHBHHx膜及添加5%、10%中药涂层的材料表面的人脐静脉内皮细胞活力旺盛、形态饱满、分布均匀。结论经表面改性处理的、且加入5%、10%中药涂层的PHBHHx膜,具有良好的人脐静脉内皮细胞相容性,在体外细胞培养的环境下,有利于细胞的生长、贴附和增殖。

Objective To evaluate the growing condition of human umbilical vein endothelial ceils （HUVECs）, which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3- hydroxybutyrate-co-3-hydroxyhexanoate copolyesters （PHBHHx） and the biocompatibility between PHBHHx tim and endothelial cell. Methods The HUVECs were harvested by Baudine method, and identified by immunohistoehemieal method. Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours, 12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluoreseence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated, and immunohistochemistry staining of FLK-1 and factor Ⅶ was positive. The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs in vitro, and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy. Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation in vitro.