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相容性

背景:聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片材料在构建组织工程支架上具有良好的应用前景,内皮细胞能否在该材料上存活和稳定生长将直接影响其作为载内皮细胞载体支架表面可降解材料的应用.目的:观察聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物与人脐静脉内皮细胞的相容性.设计;随机对照观察.单位:吉林大学第二临床医学院.材料:本实验于2006-02/10于吉林大学基础医学院病理生理教研室完成.长约20 cm脐带来自吉林大学第二医院妇产科1名正常足月分娩的新生儿,标本采集经新生儿家属知情同意,本实验经过医院伦理委员会批准.聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片由中国科学院长春应用化学研究所提供.倒置显微镜及相差显微镜为日本Olympus公司产品.方法:将从脐带中分离培养的稳态生长的人脐静脉内皮细胞接种于聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片上培养作为实验组,对照组为未放聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的培养皿.①以相差显微镜下观察细胞生长情况评价人脐静脉内皮细胞与聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的细胞相容性.②采用MTT法检测细胞种植后1,3,5及7 d的细胞增殖指数.主要观察指标:①人脐静脉内皮细胞与聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的细胞相容性.②细胞种植后1,3,5及7 d的细胞增殖指数.结果:①人脐静脉内皮细胞与聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的细胞相容性:相差显微镜下种植在聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片上的细胞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,经多次传代后,细胞呈长梭形或多角形,与对照组相比无明显差异.扫描电镜下观察在聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片上培养15 d的人脐静脉内皮细胞嵌入膜片间隙生长,但没有连接成片铺在膜片上.②细胞增殖指数:MTT法检测结果显示实验组及对照组实验后1,3,5及7 d的细胞增殖指数差异无显著性意义(P＞0.05).结论:内皮细胞在聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物上生长良好,两者具有良好的细胞相容性.

Cytocompatibility of biodegradable poly(ethylene glycol)-b-poly(L-lactide)-b-poly(L-glutamic acid) tri-block copolymer with human umbilical vein endothelial cells

BACKGROUND: Poly(ethylene glycol)-b-poly(L-lactide)-b-poly(L-glutamic acid) (PEG-PLA-PGL) tri-block copolymers have good applied foreground in constructing tissue engineering scaffold materials. Whether endothelial cells survive and grow on the materials has a direct influence on the application as a biodegradable material for the scaffold of endothelial cell vector.OBJECTIVE: To explore the cytocompatibility of PEG-PLA-PGL tri-block copolymers with human umbilical vein endothelial cells (HUVECs).DESIGN: Randomized control observation.SETTING: the Second Hospital of Jilin University.MATERIALS: The experiment was carried out in the Department of Pathobiology, School of Basic Medical Sciences, Jilin University from February to October in 2006. Human umbilical cord about 20 cm length came from one neonatal infant who was delivered normally after enough months in the Department of Gynecology and Obstetrics, the Second Hospital of Jilin University. Human umbilical cord was sampled in the informed consents of the infant's family member. The experimentation was authorized by the medical ethic committee of the hospital. PEG-PLA-PGL membranes were provided by Changchun Institute of Applied Chemistry, Chinese Academy of Sciences. Inverted microscope and phase-contrast microscope were bought from Olympus Company (Japan).METHODS: HUVECs cultivated and grew steadily, were inoculated onto PEG-PLA-PGL membranes, serving as the experiment group. While the culture medium without PEG-PLA-PGL membranes were taken as the control group.①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs was evaluated by observing cellular growth through phase-contrast microscope.②The proliferation index of cells was detected by MTT method in 1, 3, 5 and 7 days after inoculation.MAIN OUTCOME MEASURES: ①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs;②The proliferation index of cells in l, 3, 5 and 7 days after inoculationRESULTS: ①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs: The observation result of phase contrast microscopy showed that, endothelial cells planted on the PEG-PLA-PGL membranes began to attach and stretch after being planted 4-6 hours. Three days later, cells grew in colonies rapidly, after 5 days, colonies began to fuse and seemed like cobble-stone. The cells were shuttle or polygon in shape after passages. There were no significant differences between the experiment and control group. Cells cultured on PEG-PLA-PGL membranes for 15 days grew in inserts with membranes, but they didn't grow into patches through scanning electron microscope.②The proliferation index of cells: No significant differences of the proliferation index of cells were detected by MTT method in 1, 3, 5 and 7 days after inoculation between experiment group and control group (P＞0.05).CONCLUSION: Endothelial cells grow well in PEG-PLA-PGL membranes, and the two have good cytocompatibility.