摘要
目的:研究羟乙基淀粉的最新剂型HES 130/0.4对器官培养保存角膜的持续脱水效果。方法:20对配对兔角膜,一半于含10%HES 130/0.4的ACF培养液中保存28d作为实验组,不再单独脱水;另一半于ACF培养液中保存28d后再葡聚糖T500脱水48h作为对照组。内皮细胞活性和植片质量评估指标包括:内皮细胞计数、角膜厚度和含水量、角膜透明度和后弹力层皱褶程度、内皮层中肌动蛋白微丝(filament actin,F-actin)的表达,及透射电镜下内皮细胞超微结构的变化。结果:保存结束时,实验组植片明显较薄,角膜透明度和皱褶程度也优于对照组,内皮细胞密度为2371±159个/mm^2。对照组继续脱水后,角膜厚度、含水量和透明度与实验组间差别变小,内皮细胞密度却降至2138±182个/mm^2。免疫印迹法证实F-actin在两组内皮层都有表达,实验组的F-actin表达水平更高。电镜下实验组的细胞超微结构改变较小。结论:HES 130/0.4的细胞毒性小,可成为器官保存液中的持续添加组分,不仅避免角膜过度水肿,还简化了保存程序,减轻了感染风险,有希望成为新型角膜脱水剂。
AIM：To investigate the fe asibility to use a new dilation agent, hydroxyethyl starch （ HES ） 130/0.4, as a new deswelling additive in cornea organ-c ulture.
METHODS：A half of 20 pairs of rabbit corneas just were cultured in ACF medium with a supplement of 10% HES 130/0.4 f or 28d as the experimental group.The control was cultured in ACF for 28d and then dehydrated in 5%dextran T500 for 48h.The e valuation parameters included the endothelial cells viability, the mean corneal central thickness before and after pr eservation, the mean water content after storage, the corneal transparency and folding, F-actin expression of corneal e ndothelium using Western blotting, and the electron microscopy observation of corneal endothelial cells.
RESULTS： A fter storage, the mean endothelial cell density of the HES 130/0.4 experimental group was 2 731 ± 159/mm 2. The experimenta l group also exhibited a thinner corneal thickness, better transparency and less folding compared with the control. After additional dehydration, the control became thin and transparent, however, its final endothelial cell density decreased greatly to 2 138 ±182/mm2 . F-actin could be seen in corneal endothelium of two groups using Western blotting, whose expression level achieved higher in the experiment group.The ultrastructure of endothelial cells of the expeir ment group remained good compared with normal cornea.
CONCLUSION：HES 130/0.4 has low toxicity, and is well tolerated for endothelial cells and can be used as a continuous supplement during organ culture. It also avoids excessive cornea swelling, simplifies the storage steps, reduces infection risk, and appears to be an alternative deswelling additive in cornea organ-culture.
出处
《国际眼科杂志》
CAS
2014年第1期13-16,共4页
International Eye Science
关键词
角膜内皮细胞
器官培养法
羟乙基淀粉
脱水
corneal endothelial cells
organ culture
hydroxyethyl starch
dehydration
