羟丁基壳聚糖携载CD133抗体的复合涂层对猪髂动脉内皮细胞的影响

Effects of the paintcoat composited with hydroxybutyl-chitosan and CD133 antibody on the endothelial cells of swine iliac artery

目的探讨羟丁基壳聚糖携载CD133抗体的复合涂层对体外培养的猪髂动脉内皮细胞(PIEC)生长影响和可能的机制。方法将PIEC接种于裸钴铬合金片(裸金属组)、雷帕霉素药物涂层钴铬合金片(药物涂层组)及羟丁基壳聚糖携载CD133抗体的复合涂层钴铬合金片(复合涂层组)上,光镜下观察PIEC细胞形态,以CCK-8试剂盒测定细胞增殖率,以划痕法检测细胞迁移率,以Transwell小室法定量迁移细胞,以一氧化氮(NO)硝酸还原酶试剂盒、碱性成纤维细胞生长因子(bFGF)-酶联免疫吸附(ELISA)试剂盒、内皮素(ET)-放免分析试剂盒测定NO、bFGF、ET-1的含量。结果 PIEC在复合涂层组钴铬合金片上生长良好,形态正常,与裸金属组、药物涂层组比较,复合涂层组细胞增殖率[(101.42±2.93)%vs(96.76±2.92)%、(75.74±3.51)%]、细胞迁移率[(102.80±3.43)%vs(97.38±3.04)%、(81.55±4.93)%]、细胞迁移数[(103.91±4.64)%vs(97.56±3.70)%、(82.99±5.03)%]、NO[(46.75±3.53)μmol/L vs(39.70±2.51)μmol/L、(33.15±3.46)μmol/L]及bFGF[(124.35±18.07)pg/mL vs(105.46±12.62)pg/mL、(88.47±13.37)pg/mL]均升高,但与裸金属组比较,ET-1[(30.98±7.31)pg/mL vs(47.67±9.59)pg/mL]降低,差异有统计学意义(P<0.01)。结论羟丁基壳聚糖携载CD133抗体复合涂层有助于体外培养的PIEC在钴铬合金片上的增殖和迁移,有望成为支架的新型涂层材料。

Objective To investigate the effects and possible mechanism of the paintcoat composited with hydroxybutyl-chitosan and CD133 antibody on the growth of endothelial cells of pig iliac artery（PIEC） in vitro.Methods PIEC were cultured on bare cobalt-chromium alloy film（bare metal group）,rapamycin eluting cobalt-chromium alloy film（drug paintcoat group） and the paintcoat composited with hydroxybutyl-chitosan and CD133 antibody eluting cobalt-chromium alloy film（composite paintcoat group）.Light microscope was introduced for morphological observation;while cell counting kit-8（CCK-8）,scratch experiment and transwell chamber method were introduced for assessment of cell proliferation,cell migration and migrating cell count,respectively.Nitrate reductase kit,enzyme-linked immunosorbent assay（ELISA） kit and radioimmunoassay kit were used to measure the levels of nitric oxide（NO）,basic fibroblast growth factor（bFGF） and endothelin-1（ET-1）.Results Normal cells and well growth of PIEC were observed in composite paintcoat group.Compared with those in bare metal group and drug paintcoat group,in composite paintcoat group there were significant increase in cell proliferation rate,migration rate,migrating cell count,NO and bFGF.However,the level of ET-1 was significantly lower in composite paintcoat group than that in bare metal group.Conclusion The paintcoat composited with hydroxybutyl-chitosan and CD133 antibody can improve the proliferation and migration of PIEC in vitro,suggesting a new coating material for stents.