-80℃保存组织工程骨异位成骨的实验研究

Experimental study of ectopic osteogenesis of tissue engineered bone cryopreserved at-80℃

目的探讨不同保存方法对组织工程骨异位成骨能力的影响。方法以骨髓基质干细胞(marrow stromal cells,MSC)复合部分脱蛋白骨培养制备组织工程骨,分别用添加或不添加冻存保护剂的保存液于-80℃深低温冻存3个月,3个月后复温。选择40只新西兰白兔,将材料植入动物脊柱旁的肌肉内,根据植入不同的材料实验分为A组,植入添加冻存剂保存的组织工程骨;B组,植入未添加冻存剂保存的组织工程骨;C组,植入未行低温保存的组织工程骨;D组,植入部分脱蛋白骨。术后4、8周取材,行组织学观察和计算机图像分析。结果术后4、8周,各组异位成骨组织学评估,A组与C组比较未见差异(P>0.05),A组或C组分别与B组比较,A和C组异位成骨能力均好于B组(P<0.001,P<0.05);D组材料无异位成骨。结论选择适宜的冻存保护剂对组织工程骨的生物活性有一定的保护作用。

Objective To study the difference of ectopic osteogenesis for tissue engineered bone cryopreserved by various methods. Methods MSC were cocultured with partialy deproteinized bone to produce tissue engineered bone. The tissue engineered bones had been cryopreserved at -80℃ with or without cryopreservation medium for three months and then thawed. The materials were implanted in paravertebral muscle of 40 New Zealand white rabbits which were divided into A, B, C,and D groups according to different transplant materials. For group A, the muscle was implanted by tissue engineered bone stored in preservation solution with cryopreservation medium. In group B, the muscle was implanted by tissue engineered bone stored in preservation solution without cryopreservation medium. For group C, the muscle was implanted by tissue engineered bone without cryopreservation. The muscle was implanted by partialy deproteinized bone in group D. When the samples were harvested at 4, 8 weeks postoperatively, the histomorphologieal and computerized graphical analysis were carried out. Results There was no difference between the group A and C （P 〉 0.05 ） , according to. histomorphologieal evaluation. The comparision study showed that ability of ectopic osteogenesis was ranged as A and C groups being better than group B （P 〈 0. 001 ,P 〈 0.05） . There was no ectopic osteogenesis in D group. Conclusion The choice of proper cryopreservation solution could optimize the bioaetivity of tissue engineered bone.