【作者】 朱明华； 曾怡； 刘程俊； 王林； 彭蔷； 秦廷武； 杨军； 车茜；

【Author】 ZHU Minghua1,WANG Lin 1,LIU Chengjun 2,ZENGYi1 PengQiang1,QINTingWu2,YANGJun1,Che Qian1 (1.Sichuan Center for Disease Control And Prevention,Chengdu 610041,China;2.West China Hospital of Sichuan University,Chengdu 610041,China)

【机构】 四川省疾病预防控制中心； 四川大学华西医院；

【摘要】 目的:观察分析玻璃化冷冻保存组织工程化人工肌腱动物体内移植后的超微结构变化特征,探索组织工程化人工肌腱体内移植后的生物学反应和肌腱修复效果,为组织工程化人工肌腱的玻璃化冷冻保存和应用提供重要的理论和实验依据。方法:将体外动态构建的组织工程化人工肌腱植入材料,经玻璃化冷冻保存后植入大鼠体内修复跟腱缺损,并与未经玻璃化冷冻保存的人工肌腱材料组进行比较,移植后2周、4周、6周及8周,分别取出移植肌腱及周围组织,经固定后作扫描电镜和透射电镜观察。结果:玻璃化冷冻保存和非冷冻保存组织工程化人工肌腱材料两组扫描电镜观察,2周时材料呈网状结构,材料间可见幼稚的肌腱细胞。6周时材料已被降解吸收,由新生的肌腱细胞和胶原纤维所替代。8周时已形成较致密的肌腱组织,肌腱细胞及胶原纤维排列均匀,胶原纤维密度随时间的延长明显增加。透射电镜观察,2周时肌腱细胞增生活跃,多数为幼稚细胞,核膜完整,核仁清晰,细胞器丰富,胶原纤维少。6周时肌腱细胞较成熟,细胞体积小,核染色深,周围可见大量胶原纤维,纤维结构较完整,横纹清晰,两组比较无明显差异。结论:玻璃化冷冻保存和非冷冻保存组织工程化人工肌腱材料两组,体内移植后的电镜观察结果基本一致,新生的肌腱细胞和胶原纤维组织生长良好,其形态和排列与正常肌腱组织相似。证明玻璃化冷冻保存人工肌腱的方法对肌腱细胞功能无明显影响,植入的肌腱细胞具有明显的存活和生长繁殖的功能,新生的肌腱组织具有良好的生长和修复功能。更多还原

【Abstract】 Objective:To observe the feature of ultrastructure changes in the tissue engineered artificial tendon of vitreous cryopreservation,investigate the biological reaction and repairing effect of tendon after an implantation in vivo to provide an important theoretical and experimental basis for the vitreous cryopreservation and application of tissue engineered artificial tendon.Methods:After vitreous cryopreservation,implantation materials of tissue engineered artificial tendon dynamically constructed in vitro were implanted into rats to repair the tendon defect,and were compared with non-vitreous cryopreservation artificial tendon materials.Two,four,six and eight weeks after the implantation,the tendon implanted and surrounding tissues were withdrew to be fixed for the observation using scanning and transmission electron microscopes.Results:Tissue engineered artificial tendon materials of vitreous cryopreservation and non-vitreous cryopreservation in two groups were observed using a scanning electron microscope.At the 2nd week,materials presented a reticular formation and there were juvenal tendon cells among materials.At the 6th week,materials had been degraded and absorbed,and then be substituted by neonatal tendon cells and collagen fibers.At the 8th week,dense tendon tissues containing uniform tendon cells and collagen fibers had been formed; the density of collagen fibers significantly increased with time.Observing using a transmission electron microscope at the 2nd week,tendon cells proliferated actively,most of them were immature cells with a complete nuclear membrane,clear nucleolus and little collagen fibers; at the 6th week,tendon cells were more mature with a little size,deep stained nucleolus surrounded by plenty of collagen fibers with complete fiber structure and clear cross striation.There was no significant difference between two groups.Conclusion:Using an electron microscope,a very good agreement in observation was found between tissue engineered artificial tendon materials of vitreous cryopreservation and non-vitreous cryopreservation after the implantation in vivo in two groups.Neonatal tendon cells and collagen fiber tissues grew well in a similar form and order to normal tendon tissues.This proved that vitreous cryopreservation has no significant influence on the function of tendon cells,tendon cells implanted had a significant function of survival,growth and reproduction,and neonatal tendon tissues had a good function of growth and repair.更多还原