Different Concentrations Of Trehalose Cryopreserved In Liquid Nitrogen Deep With Valved Main Artery Tissue Protective Effect And Its Effect On The Expression Of Caspase-3

Objective To observe the effect of trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen on their tissues and seek the best protective agents and their best concentrations, otherwise, detect the expression of caspase-3.Methods The aortic valve homograft was divided into 5 groups, namely:0.1mol/L DMSO (control group), O.lmol/L trehalose (experimental group 1), 0.1mol/L trehalose +0.1mol/L DMSO (experimental group 2),0.2mol/L trehalose+0.1mol/L DMSO (experimental group 3),0.3mol/L trehalose+0.1mol/L DMSO (experimental group 4). At the time of 12 months,15 months and 18 months after preserved in liquid nitrogen, the expression of caspase-3 of the aortic valve homograft was measured by the light microscopy RT-PCR and Western Blot. At the same time, the glucose consumption of each group in 24 Hours was detected to represent the cell metabolism. Fresh group was a negative control group.Results The longer of the samples stored, the worse of the tissues and cells destroyed. In the light microscopy, fresh aortic tissue was dominant. Compare with the fresh group, each group had statistical significance (d=37～43, P<0.05).Among groups, there was significant difference (d=0～42, P<0.05)except the experimental group 2 and 3(d=66～72, P>0.05). The experimental group 2 was in accord with the experiment group 3, which was of a sort compare with the fresh group. The experimental group 4, which was worse than the experimental group 2 and 3, ranked the experimental group l.The worst was the control group. Dextroglucose-metabolized mensuration of the homograft:among groups, there was significant difference(F=41～46, P<0.05) other than the experimental group 2 and 3 (P=0.68> 0.05). The experimental group 2 was the same as the experiment group 3, which was modest quality compare with the fresh group. The experimental group 4, which was lower than the experimental group 2 and 3, was superior to the experimental group 1. The lowest was the control group. In RT-PCR and Western Blot, at the same time(P<0.05), the expression of caspase-3 of fresh aortic tissue was slightest. The experimental group 2 was in accord with the experiment group 3, which was of a sort compare with the fresh group. The experimental group 4, which was worse than the experimental group 2 and 3, ranked above the experimental group 1.The worst was the control group.Conclusion The trehalose as the cryoprotectant for preserving aortic valve homograft in liquid nitrogen had a good protective effect. The separate use of trehalose was better than the separate use of DMSO. The joint use of trehalose and DMSO was better than the separate use of either one. The best concentrations of trehalose of the joint use were between 0.1 mol/L and 0.2mol/L.