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| 壳聚糖和磷脂化壳聚糖涂层膜对血管内皮细胞生长和血液相容性的影响 |
| 刘宗军,孟晟,秦永文,钟伟,金惠根,于文,罗启剑 |
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| (第二军医大学长海医院心内科,上海,200433;复旦大学高分子科学系,聚合物分子工程教育部重点实验室,上海,200433;上海中医药大学附属普陀医院心内科,上海,200062;复旦大学附属华山医院药剂科,上海,200040) |
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| 摘要: |
| 目的:观察聚合物壳聚糖(chitosan,CHI)和磷脂化壳聚糖(phosphorylcholine chitosan,PC-CHI)涂层膜对培养的血管内皮细胞增殖和迁移及血液相容性的影响.方法:将CHI和PC-CHI均匀喷涂在培养皿底层制成聚合物膜,以316 L不锈钢片做成不锈钢圆柱体槽,将猪髂动脉内皮细胞接种于聚合物膜、不锈钢槽和不做任何处理的培养皿底部(空白对照组),培养24 h.光镜和扫描电镜观察细胞形态,以CCK-8试剂盒测定细胞增殖,并进行细胞迁移检测.以凝固法测定健康人血液在聚合物膜上作用2 h后的凝血活酶时间(APTT)、凝血酶原时间(PT)、纤维蛋白原测定 (FiB )和凝血酶时间(TT).结果:动脉内皮细胞在CHI和PC-CHI膜上生长良好,形态正常.培养24 h,内皮细胞在CHI和PC-CHI膜上的增殖率分别达88.8﹪和 77.8﹪,存活内皮细胞数目较不锈钢片组显著增加(P＜0.01);而CHI组的存活内皮细胞数目显著高于PC-CHI组(P＜0.01).培养72 h,内皮细胞在CHI和PC-CHI膜上迁移细胞数均显著高于316 L不锈钢片,PC-CHI组较CHI组的细胞迁移数目显著增加(P＜0.01).PC-CHI组、316 L不锈钢片组和空白对照组的APTT较CHI组显著延长(P＜0.05,P＜0.01),FiB显著增加(P＜0.01).各组间PT和TT均无明显差异.结论:CHI和PC-CHI涂层膜有助于血管内皮细胞的增殖和迁移,对内皮细胞的相容性良好.PC-CHI较CHI有显著抗血栓形成作用. |
| 关键词: 壳聚糖、磷脂化壳聚糖、血管、内皮细胞、细胞增殖、细胞运动 |
| DOI：10.3724/SP.J.1008.2007.00019 |
| 投稿时间：2006-08-22修订日期：2006-11-27 |
| 基金项目: |
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| Effects of chitosan and PC-chitosan-coated membranes on growth and hemocompatibility of vascular endothelial cells |
| LIU Zong-jun,MENG Sheng,QIN Yong-wen,ZHONG Wei,JIN Hui-gen,YU Wen,LUO Qi-jian |
| (第二军医大学长海医院心内科,上海,200433;上海中医药大学附属普陀医院心内科,上海,200062;复旦大学附属华山医院药剂科,上海,200040) |
| Abstract: |
| Objective：To investigate the effects of chitosan （CHI） and PC-chitosan （PC-CHI）-coated membranes on the proliferation, migration, and hemocompatibility of vascular endothelial cells. Methods： CHI and PC-CHI were separately sprayed on the bottom of culture dish evenly to form the polymer membrane; the cylinder was made of 316 L stainless steel flake. Pig iliac endothelial cells （PIEC） were cultured on the polymer membranes, the stainless steel bottom of the cylinder, and the untreated bottom of culture dish （as blank control）. Then the cell morphology was observed by light microscope and scan electron microscope; cell proliferation was measured by Cell Counting Kit-8 （CCK-8）; and cell migration was also assayed. Whole blood activated partial thromboplastin time （APTT）, prothrombin time （PT）, fibrinogen （FIB）, and thrombin time （TT） were examined by clotting method. Results： PIEC grew well on the membranes of CHI and PC-CHI, with normal morphology. After 24 h culture, the proliferation rates of PIEC were 88.8% and 77.8% on CHI and PC-CHI membranes, respectively. The survived PIEC on CHI and PC-CHI membranes were significantly more than those on 316 L stainless steel flake（P〈0.01）, and those on CHI membrane were significantly more than those on PC-CHI membrane（P〈0.01）. After 72 h culture, the numbers of migration cells on CHI and PC-CHI membranes were significantly more than those on 316 L stainless steel flake（P〈0. 01）, and those on PC-CHI membrane were significantly more than those on CHI membrane（P〈0.01 ）. APTT in PCCHI, 316 L stainless steel flake, and blank control groups were significantly longer than that of C HI group（P〈0.01, P〈0.05）, and FiB levels in the former 3 groups were higher than that of CHI group（P〈0.01 ）. Conclusion.. Coating with CHI and PC-CHI can improve the proliferation and migration of vascular endothelial cells on the culture dishes, indicating an excellent biocompatibility of CHI and PC-CHI membranes. PC-PCH membrane has more potent antithrombotic effect than CHI does. |
| Key words: chitosan PC-chitosan blood vessels endothelial cells cell proliferation cell movement |
