液氮超低温保存同种带瓣动脉活性实验研究

Experimental study on the viability of cryopreserved allograft valved aortic and pulmonary arteries

目的 探讨活细胞同种瓣的保存方法和应用 ,观察同种带瓣动脉的消毒和冷冻保存对动脉瓣膜活性的影响。方法 应用抗生素消毒、液氮超低温保存同种带瓣动脉 36个。采用O′Brien氏抗菌素灭菌法 ,速率降温后转入液氮液相中长期保存。解冻后进行细菌学检查、形态学检查 ,冷冻前、解冻后 ,均取部分二尖瓣和主动脉壁分别做糖代谢测定。其中 2个主动脉瓣和 1个肺动脉瓣分别于灭菌前 ,结冻后取样做电镜检查。结果 光镜检查 :灭菌后瓣叶及动脉壁组织大体结构 ,细胞形态无明显变化。带瓣动脉解冻后瓣叶组织细胞核清晰 ,弹性纤维结构完整 ,有轻度灶性粘变性和少数内皮细胞脱落 ,主动脉壁结构大致正常。电镜检查 :灭菌后及解冻后组织大多数细胞核清晰 ,核膜完整 ,染色质分布正常 ,线粒体和内质网轻度肿胀 ,线粒体嵴规则 ,偶见变性或坏死的细胞。糖代谢测定 :灭菌后动脉壁和二尖瓣组织葡萄糖消耗率平均为 54～56mg/dl/ 2 4h ;解冻后动脉壁、二尖瓣和主动脉瓣组织葡萄糖消耗率平均为 51～ 71mg/dl/ 2 4h。结论 抗生素消毒液氮超低温保存同种带瓣动脉获得了组织良好的活性 ,有自然结构 ,最佳血流动力学 ,无支架结构 。

Objective To search for preserved method of viable allograft valved aortic and pulmonary arteries.And to investigate the influence of sterilization and preservation to viability of allograft valved aortic and pulmonary arteries. Methods 36 of allograft valved aortic and pulmonary arteries was prepared adopting O′Brien MF′s sterilized with antibiotics and cryopreserved in liquid nitrogen for 3～24 months.The bacteriological and morphological determining and dextroglucose-metabolized mensuration of the homograft precryopreserved,after-resuscitated were studied by germiculture and electron microscope and tissue culture. Results The integrity of the homografts cryopreserved in liquid nitrogen was kept intact.Cellular morphosis has not changed.The nucleuses are normal.The structure of elastic fibres is integrity.The distributing of chromatin is normal.Mitochondrion and crista and endoplasmic reticulum are regulation and normal basically.Dextroglucose-metabolized mensuration of the homograft:The rate of consume precryopreserved was 54～56 mg/dl/24 hour in average,The rate of consume after-resuscitated was 51～71 mg/dl/24 hour. Conclusion allografts valved aortic and pulmonary arteries cryopreserved in liquid nitrogen for 3～24 month have excellent viability with natural biophysical structure for transplantation and it is an ideal vascular substitute as out conduit in the surgical treatment of complex cardiac malformation and as transannular patch to repair right ventricular outflow tract.