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应用犬真皮成纤维细胞构建组织工程半月板修复犬半月板缺损的实验研究
Tissue Engineered Meniscus Constructed by DFs in the Repair of Meniscus Defects
【摘要】 目的探讨体外构建组织工程半月板修复半月板缺损的可行性。方法将体外扩增至第4代的犬真皮成纤维细胞(Dermal fibroblasts,DFs)接种于PGA/PLA支架材料上,应用软骨形态发生蛋白1(Cartilage-derived morphogeneticprotein-1,CDMP1)细胞诱导液体外诱导培养2周后,回植于犬自体半月板缺损模型体内,移植后6个月取材观察半月板的再生情况。结果诱导组模型内形成较大的新生半月板组织,组织学染色显示诱导组新生组织更接近于半月板组织的生理特点,尤其是内侧部分大部分细胞表现出软骨细胞特有的陷窝结构;非诱导组及空白对照组新生的组织较小,且组织学表现更类似纤维结缔组织。新生半月板Ⅰ型胶原偏振光检测显示,诱导组胶原纤维排列较非诱导组致密,出现Ⅰ型胶原的横纹特征。扫描电镜检测显示,诱导组细胞外基质与正常半月板相比较疏松,非诱导组细胞外基质则更加疏松。膝关节胫骨平台关节软骨的HE染色显示,诱导组关节面的外侧有新生半月板组织覆盖的部位关节软骨尚有部分存余,而诱导组已基本消失。结论组织工程化细胞材料复合物能在犬自体半月板缺损模型上实现半月板再生。
【Abstract】 Objective To explore the feasibility of using tissue engineered meniscus to repair meniscus defects.Methods DFs isolated from canine and expanded to the fourth passage were seeded in PGA/PLA scaffold and induced with CDMP1 for 2 weeks,then implanted to repair defects of autologous medial meniscus.Six months after implantation,the new formed meniscus were harvested for detection.Results The constructs of CDMP1 induced group formed cartilage-like tissue,and most cells demonstrated a typical morphological characteristic of chondrocyte especially in the inner edge.Whereas the noninduced group generated smaller and more fibrous like tissue,and only a few cells present lacuna like structure.Polarizing microscope examination showed that the bundles of collagen type Ⅰ in CDMP1 induced group was thicker than that in noninduced group.SEM evaluation demonstrated more dense ECM in CDMP1 induced group compared with non-induced group,but significantly less than normal meniscus.HE staining of newly formed meniscus on tibial plateau showed more degree of protection for covered hyaline cartilage in CDMP1 induced group compared with non-induced group.Conclusion Tissue engineered constructs can further improve meniscus regeneration to repair the autologous meniscus defects.
【Key words】 Dermal fibroblasts; Meniscus; Cartilage-derived morphogenetic protein-1; Tissue engineering;
- 【文献出处】 组织工程与重建外科杂志 ,Journal of Tissue Engineering and Reconstructive Surgery , 编辑部邮箱 ,2013年02期
- 【分类号】R329;S858.292
- 【下载频次】126