快速冷冻与慢速冷冻条件下复合组织血管内皮的生物活性  

Biological activity of vascular endothelial cells of composite tissue cryopreserved with rapid-freezing versus slow-freezing methods

作　　者：刘元新[1] 张树明[2] 乔林[2] 霍海涛[2] 朱泽兴[2] 袁磊[2] 

机构地区：[1]辽宁医学院研究生学院,辽宁省锦州市121000 [2]解放军第二炮兵总医院骨科,北京市100088

出　　处：《中国组织工程研究》2012年第20期3659-3662,共4页Chinese Journal of Tissue Engineering Research

摘　　要：背景:恢复良好血供对于复合组织保存及再植至关重要,但不同冷冻方法对血管活性影响不同。目的:比较快速冷冻与慢速冷冻方法对复合组织血管内皮活性的影响。方法:新西兰大白兔后肢分别进行快速冷冻与慢速冷冻,快速冷冻的兔后肢直接放入液氮中,慢速冷冻组经4℃,-20℃,-80℃冷冻后再投入到液氮中,各组依次保存12h,3d,7d后快速复温,并设对照组。所有新西兰大白兔后肢均经甲醛溶液固定后行苏木精-伊红染色及免疫组织化学染色检测各组血管内皮的病理变化。结果与结论:快速冷冻组和慢速冷冻组冷冻12h,3d,7d时兔血管组织形态评分均低于对照组(P<0.05)。快速冷冻组冷冻12h,3,7d时血管内皮组织血管内皮生长因子评分低于慢速冷冻组(P<0.05)。证实,慢速冷冻法能更好的维持复合组织中的血管内皮细胞的生物学活性。BACKGROUND： To recover good blood supply is critical to the cryopreservation and replantation of composite tissues, and different cryopreservation techniques have different effects on vascular activity. OBJECTIVE： To compare the activity of vascular endothelial cells of composite tissue following rapid-freezing and slow-freezing methods. METHODS： Forty New Zealand white rabbits were randomly divided into control group, cryopreservation for 12 hours group, 3 days group, 7 days group, with 10 in each group. Control group was without any treatment. In the experimental groups, New Zealand white rabbit hind limbs were divided into two subgroups, receiving rapid freezing and slow freezing. The hind limbs in the rapid freezing group were put into liquid nitrogen for 12 hours, 3 days, and 7 days. In the slow-freezing group, the freezing procedure was carried out at 4 ℃, -20 ℃ and -80℃, successively, using programmed frozen. All the groups required cryopreservation for 12 hours, 3 days, and 7 days, and then rapid rewarming. After thawing, pathological changes in the vascular endothelium were examined by optical microscope and immunohistochemical staining. RESULTS AND CONCLUSION： The scores on the morphology of vascular endothelial tissues were higher in the control group than the two freezing groups following cryopreservation for 12 hours, 3 days, and 7 days （P 〈 0.05）. The scores on vascular endothelial growth factors were higher in the slow-freezing group than the rapid-freezing group （P 〈 0.05）. These findings indicate that slow-freezing is superior to rapid-freezing to preserve the biological activity of vascular endothelial cells of composite tissue.

关 键 词：深低温冷冻 复合组织 冷冻方法 内皮细胞活性 组织构建 

分 类 号：R318[医药卫生—生物医学工程]