地塞米松对深低温保存猪角膜内皮细胞的影响

Study on the influences of dexamethasone to cryopreserved cornea endothelial cells

目的:研究地塞米松预处理对液氮深低温冻存猪角膜内皮细胞的影响。方法:选择新鲜猪角膜,以13mm直径环钻制取角膜标本,依次放入4种冷冻保护液中冷平衡,"简化四步法"程序降温后放入液氮中保存,使用前40℃水浴复温。地塞米松作用组以同法制备植片,前三步冷平衡同前,最后分别放入含0.01,0.03,0.05mg/mL地塞米松保护液中4℃孵育18h,再程序降温液氮冻存。复温后用于锥兰-茜素红染色,观察内皮细胞存活情况。结果:内皮细胞染色结果示:程序冻存组和地塞米松作用组的内皮细胞存活率均超过90%,差异比较无统计学意义(P>0.05)。结论:在0.01～0.05mg/mL浓度范围内,未见地塞米松毒性作用,内皮细胞维持正常六角形形态,内皮细胞存活率超过90%。

AIM： To study the influences of dexamethasone to cryopreserved cornea endothelial cells. METHODS： Fresh pig corneas were drilled down by 13mm-diameter trephine. Then the grafts were put into four kinds of cool cryoprotective media by turns and equilibriumed with ＇ simplified four - step＇ method. All grafts were thawed in 40℃. All dexamethasone pretreatment corneas were additionally incubated in the last cool cryoprotective medium which had 0.01mg/mL, 0.03mg/mL or 0.05mg/mL dexamethasone for 18 hours in 4℃ before cryopreservation. Corneas thawed were stained by typan blue and alizarin Bordeaux. Survival rates of endothelia cells were observed. RESULTS： There were no significant differences in endothelia survival rate among rate-freezing group and every dexamethasone pretreetment group. CONCLUSION： In 0.01 -0.05mg/mL of concentrations, dexamethasone shows no toxic effects. Survival rate of endothelial cells was more than 90%.