壳聚糖体外降解及降解产物对内皮细胞增殖的影响.docVIP

壳聚糖体外降解及降解产物对内皮细胞增殖的影响* 【摘要】 目的 研究壳聚糖体外降解的规律及不同降解时间其产物对血管形成密切相关的内皮细胞增殖的影响，为壳聚糖在组织工程血管化方面的应用提供参考。方法 以生理盐水为降解介质对壳聚糖膜进行体外降解，用失重率、黏均分子量变化和FTIR表征降解本体的变化规律，用降解液pH值和降解产物中氨基葡萄糖浓度变化表征降解产物的变化规律；将降解液与内皮细胞共培养，用MTT法检测内皮细胞增殖情况，并与氨基葡萄糖单体培养后内皮细胞增殖结果进行比较。结果壳聚糖膜在生理盐水中的降解是壳聚糖分子链中β(1, 4) 糖苷键断裂引起的，降解缓慢，120 d失重率为19.48％，降解过程中黏均分子量减小，最终产物为氨基葡萄糖，其浓度不断增大，降解产物的累积使pH值升高。不同时间降解产物中氨基葡萄糖浓度都在氨基葡萄糖单体促进内皮细胞增殖的范围内，前期和后期的降解产物促进内皮细胞增殖，但降解中期21～90 d的降解产物对内皮细胞的增殖有抑制作用，表明氨基葡萄糖单体并非影响内皮细胞增殖的主要因素。结论 壳聚糖体外降解缓慢，降解最终产物为氨基葡萄糖，其降解产物对内皮细胞增殖在降解中期抑制，前期和后期促进，其主要影响因素不是氨基葡萄糖单体。本研究可为壳聚糖在组织工程血管化方面的应用提供参考。 【关键词】 壳聚糖；体外降解；降解产物；内皮细胞 Degradation of Chitosan in Vitro and Effects of Degradation Products on Proliferation of Endothelial Cells. SHI Guoqi,CHEN Yuanwei,DING Yulong,YU Xixun,ZHANG Xiaohua,WAN Changxiu. Abstract： Objective To study the degradation of chitosan in physiological saline and the effect of degradaion products on proliferation of endothelial cells, as well as to provide the reference for exploring the potential application of chitosan to promote angiogenesis in tissue engineering. Methods The degradation was prepared by immersing the films of chitosan in physiological saline under aseptic condition. The weight loss rate and molecular weight were measured. On the other hand, the pH value and glucosamine concentration of degradation products were measured with MorganElson method. ECV304, a human umbilical vein endothelial cell line, was used and cultured with the degradation fluid. Cell proliferation was detected with MTT assay. The fluids of glucosamine with different concentrations also were cultured and compared with ECV304. Results The degradation of chitosan went along by destroyed the beta;(1, 4) bond. The weight loss of chitosan was 19.48% in 120 d. The molecular weight decreased with the increasing of degradation time. The pH value increased slightly and the concentration of glucosamine increased always with the increasing of degradation time. The degradation products of 21～90 d inhibited