玻璃化冷冻保存组织工程肌腱修复跟腱缺损的超微结构影响研究  

Research on Influence of Repair with Tissue Engineered Tendon of Vitreous Cryopreservation on Ultrastructure of Achilles Tendon Defect

作　　者：朱明华[1] 王林[1] 刘程俊[2] 曾怡[1] 秦廷武[2] 王睿[1] 朱锐[1] 杨军[1] 车茜[1] 

机构地区：[1]四川省疾病预防控制中心,成都610041 [2]四川大学华西医院骨科,成都610041

出　　处：《生物医学工程学杂志》2010年第3期590-594,共5页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30570469)

摘　　要：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,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。将玻璃化冷冻保存后的组织工程化人工肌腱植入大鼠体内修复跟腱缺损,移植后2、4、6、8周分别取出移植肌腱及周围组织。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。玻璃化冷冻和非冷冻保存两组超微结构观察结果基本一致,新生的肌腱细胞和胶原纤维组织生长良好,形态结构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,新生的肌腱组织具有良好的生长和修复功能。By observations of the features of ultrastructure changes in the tissue engineered artificial tendon of vitreous cryopreservation,we investigated the repairing effect of tendon after an in-vivo implantation and hence we provided an important theoretical and experimental basis for the vitreous cryopreservation and application of tissue engineered artificial tendon. After vitreous cryopreservation,the implantation materials of tissue engineered artificial tendon dynamically constructed in vitro were implanted in rats for reparation of the tendon defect. A scanning electron microscope was used. At the 2nd week,the materials presented a reticular formation and there were juvenile tendon cells among materials. At the 6th week,materials were already degraded and absorbed,and then were substituted by neonatal tendon cells and collagen fibers. At the 8th week,dense tendon tissues containing uniform tendon cells and collagen fibers were found already formed; the density of collagen fibers significantly increased with time. Using a transmission electron microscope at the 2nd week,we found active proliferation of tendon cells; 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-sized,deep-stained nucleolus surrounded by plenty of collagen fibers with complete fiber structure and clear cross striation. There was no significant difference between the two groups. Using an electron microscope,we found a very good agreement in observation of the tissue engineered artificial tendon after the in-vivo implantation in two groups. Neonatal tendon cells and collagen fiber tissues grew well and are in a similar form and order when compared versus normal tendon tissues. This proved that vitreous cryopreservation has no sig-nificant influence on the function of tendon cells. The neonatal tissue-engineered tendon exerts good function of growth and repair.

关 键 词：组织工程肌腱 玻璃化冷冻保存 肌腱移植 电镜观察 

分 类 号：R318.08[医药卫生—生物医学工程] R318.1[医药卫生—基础医学]