明胶对人脐静脉内皮细胞株细胞相容性影响的研究

In vitro proliferation and morphological comparison of human umbilical vein endothelial cell cultured with three different molecular weights of gelatin hydrogel microspheres by two methods of administration

目的探讨明胶对人脐静脉内皮细胞株(HUVECs)细胞生长的影响。方法建立后干扰(模型1)及预干扰(模型2)的细胞模型,采用MTT法和倒置光镜动态观察细胞生长状态及观察不同分子量明胶(40000道尔顿、60000道尔顿、70000道尔顿)、不同质量分数(5%、10%、15%)对HUVECs细胞相容性的影响。实验分为10组:对照组,明胶Aa、Ab、Ac组(明胶分子量为40000道尔顿,明胶溶液浓度分别为5%、10%、15%),明胶Ba、Bb、Bc组(明胶分子量为60000道尔顿,明胶溶液浓度分别为5%、10%、15%),明胶Ca、Cb、Cc组(明胶分子量为70000道尔顿,明胶溶液浓度分别为5%、10%、15%)。结果 (1)倒置光镜动态观察显示:模型1:对照组HUVECs呈正常生长状态,3个时间段(24、48、72h)细胞形态及密度无明显变化,而与对照组比较,明胶各实验组细胞大量死亡,仅少数细胞贴壁存活;模型2:对照组HUVECs呈正常生长状态,与对照组比较,3个时间段(24、48、72h)内明胶各实验组细胞生长状态好,无明显差异,并于72h时出现细胞密度明显增加的现象。(2)MTT检测:模型1:与对照组比较,明胶各实验组3个时间段(24、48、72h)细胞增殖有明显降低,至72h,HUVECs增殖无明显增加;模型2:与对照组比较,随着观察时间增加,3个时间段(24、48、72h)明胶各实验组细胞增殖逐渐增加,其中48h明胶Aa组、72h明胶Cc组两组与对照组HUVECs细胞增殖比较差异有统计学意义(P<0.05)。结论在本研究范围内,采用预干扰模型(模型2)对较高分子量明胶进行细胞实验较适合;较高分子量明胶可能对微血管在一定时间内的修复与重建有促进作用。

Objective To observe the effect of different molecular weight and concentration of gelatin on cellular compatibility of human umbilical vein endothelial cells（HUVECs）in vitro experiment. Methods Establishment of pretreatment and after-treatment models and using MTT and morphological comparison of human umbilical vein endothelial cells （HUVECs） to test protective effect of cells by different molecular weight （40 000 Dalton, 60 000 Dalton and 70 000 Dalton） and different concentration of gelatin （5%, 10% and 15%）. And it was divided evenly and randomly into ten groups, including normal group,group Aa, Ab, Ac（40 000 Dalton, Concentration of gelatin 5%, 10%, 15%）, group Ba, Bb, Bc（60 000 Dalton, Concentration of gelatin 5%, 10%, 15%）, group Ca, Cb, Cc（70 000 Dalton, Concentration of gelatin 5%, 10%, 15%）. Results （1） The results by light microscopic showed that comparison of normal group, most of cells in groups of gelatin were died in the first model after 24,48,72 h. But the growth of cells and cell morphology were good in the second model for three days. And cell density of groups of gelatin were significantly increased at 72 h.（2）The result by MTT showed that comparison of normal group, proliferation of cells in groups of gelatin for three days were significantly decreased. Besides, with increasing molecular weight of gelatin, more concentration of gelatin, and more the cell inhibition（P0.01） in the first model. It was dose-dependent manner and no time-dependent manner. And With increasing molecular weight of gelatin, less concentration of gelatin, and more the cell inhibition, especially group Aa （P0.05） at 48 h and group Cb at 72 h（P0.05） in the second model. Conclusions It shows that it may be more suitable for gelatin by pro-treatment’s experiment. And the higher molecular weight of gelatin may be good for microvascular reconditioning for a long time.