蔗糖在真空冷冻干燥和储藏过程中对兔角膜内皮细胞保护作用的实验研究

The Experimental Research of Sucrose Protective Effect on Rabbit Corneal Endotheliun During Vacuum Freeze-drying

目的本文评价蔗糖在冻干兔角膜中的价值,并探讨其作用机制。方法在影响冻干兔角膜效果的其他因素固定前提下,使用二甲基亚砜、白蛋白为抗冻剂基础上加入不同浓度的蔗糖。(对照组中蔗糖浓度梯度为2.5%,5.0%,7.5%,10.0%;实验组Ⅰ中均为0;实验组Ⅱ中为73.5%,15.0%,22.5%,30.0%;实验组Ⅲ中为25.0%,50.0%,75.0%,100.0%)实验分两阶段进行即冷冻阶段和冻干阶段。通过台盼兰和茜素红联合染色评价其活性。计算平均ESR,进行统计学分析。结果冷冻阶段,对照组平均ESR为96.1%,实验组Ⅰ平均ESR为94.0%,实验组Ⅱ平均ESR为95.2%,与对照组经方差分析蔗糖为不显著因素。实验组Ⅲ平均ESR为40.7%,经方差分析蔗糖为显著因素。干燥阶段,对照组平均ESR为40.3%,实验组Ⅱ的ESR为71.1%,实验组Ⅰ、实验组Ⅲ的ESR无法计算。方差分析表明,在冻干阶段蔗糖为高显著因素。结论蔗糖在冻干兔角膜中对内皮细胞具有保护作用。在冷冻阶段蔗糖起协同保护作用。在干燥阶段起主要保护作用。蔗糖对冻干兔角膜中对内皮细胞保护作用与浓度有关,保护液中蔗糖浓度梯度为7.5%,15.0%,22.5%,30.0%时,保护效果最好。

Objective To evaluate the protective effect of sucrose on freeze-drying rabbit corneas and analyze the mechanism. Methods Other factors are fixed add variable concentration sucrose to constant DMSO and HAS. In control group,sucrose concentration was 2.5%, 5.0%, 7.5% and 10.0%. In Experimental group Ⅰ, sucrose concentration was 0; Experimental Ⅱ was 7.5%, 15.0%, 22.5%, 30.0% ; Experimental Ⅲ was 25.0%,50.0%,75.0% and 100.0%. This experimental consists of freezing and drying. Corneal endothelium viability assessment by typan blue and alizarin red. Results In freezing step,ESR of control group is 96.1% ,experimental Ⅰ is 94.0% ,Experimental Ⅱ is 95.2%. Vzeiance analysis suggests that sucroseis was no significant. Experimental Ⅲ was 40.7%. Variance analysis suggests that sucrose was significant in freezing-drying step,ESR of control group was d0.3%,Experimental Ⅱ 71.1%,ESR of Experimental Ⅰ and Ⅲ was 0%. Variance analysis suggests that sucrose was significant. Conclusion Sucrose has protective effect on rabbit corneal endothelium during freeze-drying. Sucrose plays main protective role during freezing and cooperates with DMSO and HAS protecting cornea on freezing stage. The sucroses protective effect is interrelated with its concentration. The concentration step of sucrose 7.5%, 15.0%, 22.5%, 30.0% is best.