壳聚糖体外降解及降解产物对内皮细胞增殖的影响  

Degradation of Chitosan in Vitro and Effects of Degradation Products on Proliferation of Endothelial Cells.

作　　者：史国齐[1] 陈元维[1] 丁玉龙[1] 余喜讯[1] 张小华[1] 万昌秀[1] 

机构地区：[1]四川大学高分子科学与工程学院,四川成都610065

出　　处：《航天医学与医学工程》2009年第1期62-66,共5页Space Medicine & Medical Engineering

基　　金：国家自然科学基金资助项目(3037041150472091)

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。Objective To study the degradation of ehitosan 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 ehitosan 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 umbilieal vein endothelial cell line, was used and cultured with the degradation fluid. Cell proliferation was detected with MTF assay. The fluids of glueosamine with different concentrations also were cultured and compared with ECV-304. Results The degradation of ehltosan went along by destroyed the β-（ 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 slight- ly and the concentration of glucosamine increased always with the increasing of degradation time. The degradation products of 21 -90 d inhibited the cell proliferation, but promoted the cell proliferation on other degradation time. When the concentration of glucosamine was 10 μmol/L - l retool/L, the fluid promoted the cell proliferation. The glueosamine concentration of degradation fluid was 〉 or = 10 mmol/L the cell proliferation was inhibited. Conclusion The degradation of chitosan goes along by destroyed the 13-（ 1 , 4） bond. The degradation products of 21 -90 d inhibit the cell proliferation, but promote the cell proliferation on other degradation time. The glucosantine concentration is not the only factor which influenced the endothelial cells proliferation.

关 键 词：壳聚糖 体外降解 降解产物 内皮细胞 

分 类 号：R318.08[医药卫生—生物医学工程]