摘要
①目的观察深低温保存后兔颈总动脉内皮细胞存在状态、平滑肌细胞存活与再生能力;观察并分析深低温保存后、体外培养后动脉血管壁的结构变化、血管力学性能变化.②方法获取兔子的颈总动脉并用含有1.5 mol/L 1,2-丙二醇(PROH)的低温保护剂深低温保存,复温时采用液氮中预冷的冰袋缓慢复温.以新鲜动脉作为对照.体外培养新鲜和冷冻复温后的动脉的平滑肌细胞(SMCs);同时将冷冻复温后的血管组织在体外培养6、12和24 h,以观察血管内皮细胞和SMCs的变化,以及血管壁的完整性(弹力膜或纤维的完整性)和这些血管的力学特性(弹性模量和断裂压力)的变化.③结果冷冻复温后血管SMCs在体外培养开始生长增殖所需的时间(24～36 h)几乎与新鲜血管相似,生长的速度也相似.冷冻复温后,血管内壁存留有少量的内皮细胞,平滑肌和血管壁结构没有明显变化.但是,血管力学性能显著降低(F=5.325～97.458,q=2.32～12.38,P<0.05、0.01).冷冻复温后的血管作为完整的组织体外培养后,所有的内皮细胞脱落,部分SMCs出现变性和坏死;弹力纤维和胶原出现不同程度的断裂,力学性能进一步降低(q=2.12～8.76,P<0.05、0.01).④结论本研究中应用的方法对于保护血管的SMCs是有效的,但是对于血管内皮细胞效果不好;冷冻复温过程和复温后体外培养损伤了血管壁的结构,降低了血管的力学性能.
Objective To observe the maintenance of the endothelial cells, the survival and regenerative capability of the smooth muscle cells after cryopreservation, and to observe and analyze the structure and mechanical properties of rabbit carotids after cryopreservation or in vitro culture. Methods Rabbit carotids were harvested and cryopreserved in cryoprotective medium containing 1.5 mol/L 1,2-propanediol (PROH) and thawed slowly in ice bag precooled in liquid nitrogen. Fresh carotids were taken as controls. The fresh and frozen-thawed arteries were cultured for the growth of smooth muscle cells(SMCs). The frozen-thawed arteries tissues were also cultured in vitro for 6, 12 and 24 hours to observe the changes of endothelial cells and SMCs, the integrity of vessel wall (the elastic lamellae or fibers) and the mechanical properties (elastic modulus and fracture strength) of these carotids. Results It took almost the same time (24-36 hours) for the SMCs of frozen-thawed arteries as the fresh arteries to regenerate, and their growing speeds were also similar. After freeze-thawing procedure, a small amount of endothelium cells(ECs) existed, but no obvious changes in the SMCs and the configuration were observed. However, the mechanical properties decreased significantly(F=5.325-97.458; q=2.32-12.38;P<0.05,0.01). After in vitro culture of the frozen-thawed vessel as intact tissue, all the survived endothelial cells fell off and some of the SMCs denaturalized or necrosed; and the internal elastic fibers and collagen showed various degree of cracking, and the mechanical parameters further decreased (q=2.12-8.76; P<0.05,0.01). Conclusion The protocol used in this study for the rabbit carotid cryopreservation is effective for the protection of the SMCs, but not effective for that of the endothelial cells. The freeze-thawing process and the following in vitro culture as intact tissue injure the configuration of the vessel wall and decrease the mechanical properties of the rabbit carotids.
出处
《齐鲁医学杂志》
2005年第1期8-13,共6页
Medical Journal of Qilu
基金
国家自然科学基金(NSFC59976041)
CAS百人计划资助项目(2001 2003)
关键词
兔
颈总动脉
低温保存
力学
体外研究
超微结构
rabbits
carotid artery, common
cryopreservation
mechanics
in vitro
ultrastructure
