深低温冷冻法与酶洗法制备兔组织工程同种异体气管支架研究

摘要

目的探讨深低温冷冻与酶洗法制备的兔组织工程同种异体气管支架在去除抗原性、维持生物力学及保护细胞外基质方面的效果差别。方法18只成年新西兰兔随机分为气管未处理作为对照A组，实验B组通过深低温冷冻法处理，实验C组通过酶洗法处理，各组样本数均为6。将各组标本处理后，光镜观察行HE染色后标本，戊二醛固定后电镜扫描，测量并记录气管最大拉伸力、破裂力和变异率等生物力学性能。结果光镜观察显示A组有大量完整的气管粘膜上皮细胞；实验B组可见部分气管粘膜上皮细胞；实验C组标本未见上皮细胞，且细胞核碎裂。对照A与实验B组在电镜显示，气管支架可见丰富的细胞基质，未暴露胶原纤维；实验C组见较少细胞外基质，胶原纤维暴露。组间两两比较，气管支架的最大拉伸力、最大破裂力和变异率均无统计学差异。结论综合组织学、扫面电镜和生物力学分析，应用深低温冷冻法制备的气管支架可较好的保留细胞外基质，但不能有效的去除抗原；通过酶洗法制备气管支架可以有效地去除抗原，但未能保留较完整的细胞外基质。%Objective This study mainly focused on finding out the optimal method to obtaine rabbit histological engineering tracheal matrix between cryoapplication and detergent-enzymatic. Methods 18 adult rabbits were divided into 3 groups (each n =6), and group A were not treated as control group,groupB was treated with the method of cryopreservation,group C was treated with detergent-enzymatic method. HE dyes and scanning electron microscopy were used to observe the morphology and ultrastructure of the treated tracheal matrices. The biomechanical properties including maximum tensile force, rupture force and aberration rate of the treated tracheal matrices were measured. Results There were a lot of epithelial cells in the tracheas of group A and some epithelial cells in the tracheas of group B.But the tracheas of group C could not be seen complete epithelial cells. Under scanning electron microscopy,there were abundant extracellular matrix and col agen fiber in group A, B and C. Final y, no statistical dif erences were found in the maximum tensile force, rupture force and aberration rate of al groups. Conclusion The method of cryopreservation could retain abundant extracellular matrix and col agen fiber for tracheal stent. Howevr,this way can not remove the antigen. The method of enzymatic, by which the antigen can be removed, and extracellular matrix,but it can not retain a lot of extracellular matrix.