Objective To construct an engineered scaffold-free tendon tissue using bone-marrow mesenchymal stem cells (BMSCs) pre-treated with connective tissue growth factor (CTGF) and ascorbic acid.Methods After isolated from rat bone marrow and cultured,BMSCs at P3 were treated by CTGF and ascorbic acid for 2 weeks,and a cellular sheet formed.The BMSCs cell sheets were collected,then rolled up and loaded on a 1 cm wide U-shaped spring to form tendon-like structure.The engineered tendon tissue was sutured to posterior midline of nude mice,and histological and immunohistochemical staining were performed to assess the characteristics.Results The engineered tendon tissue showed an immature tissue structure with relatively loose extra-cellular matrix.After transplanting the engineered tissue into nude mice for 12 weeks,neo-tendon tissues formed with good collagen fibril alignment and spindle shaped cells.Conclusion An engineered tendon using BMSCs and CTGF without exogenous scaffolds was constructed in this study,and it showed potentials for neo-tendon formation.This could be a potentially new approach for tendon repair and regeneration.%目的 构建一种不含支架的组织工程学肌腱替代材料.方法 分离培养骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSCs),取3代细胞加入成肌腱诱导液2周,得到细胞膜片,在回形针上缠绕得到组织工程肌腱材料.对材料进行组织形态学检测、免疫组织化学检测、裸鼠种植实验,对比正常肌腱,观察材料特点.结果 构建的材料有着不成熟的类似肌腱组织的结构,裸鼠体内种植12周后获得类肌腱组织,其细胞外基质成熟,结构成分与肌腱组织接近.结论 本实验通过BMSCs构建出无人工材料的组织工程化肌腱,这种材料作为替代材料用于肌腱修复理论上可行.
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