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腺病毒介导的兔骨髓间充质干细胞 Sox9 基因转染促进软骨缺损修复。

The promotion of cartilage defect repair using adenovirus mediated Sox9 gene transfer of rabbit bone marrow mesenchymal stem cells.

机构信息

Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China.

出版信息

Biomaterials. 2011 Jun;32(16):3910-20. doi: 10.1016/j.biomaterials.2011.02.014. Epub 2011 Mar 5.

Abstract

Although Sox9 is essential for chondrogenic differentiation and matrix production, its application in cartilage tissue engineering has been rarely reported. In this study, the chondrogenic effect of Sox9 on bone marrow mesenchymal stem cells (BMSCs) in vitro and its application in articular cartilage repair in vivo were evaluated. Rabbit BMSCs were transduced with adenoviral vector containing Sox9. Toluidine blue, safranin O staining and real-time PCR were performed to check chondrogenic differentiation. The results showed that Sox9 could induce chondrogenesis of BMSCs both in monolayer and on PGA scaffold effectively. The rabbit model with full-thickness cartilage defects was established and then repaired by PGA scaffold and rabbit BMSCs with or without Sox9 transduction. HE, safranin O staining and immunohistochemistry were used to assess the repair of defects by the complex. Better repair, including more newly-formed cartilage tissue and hyaline cartilage-specific extracellular matrix and greater expression of several chondrogenesis marker genes were observed in PGA scaffold and BMSCs with Sox9 transduction, compared to that without transduction. Our findings defined the important role of Sox9 in the repair of cartilage defects in vivo and provided evidence that Sox9 had the potential and advantage in the application of tissue engineering.

摘要

虽然 Sox9 对于软骨细胞分化和基质生成至关重要,但它在软骨组织工程中的应用却鲜有报道。本研究评估了 Sox9 对骨髓间充质干细胞(BMSCs)的体外软骨生成作用及其在体内关节软骨修复中的应用。用携带 Sox9 的腺病毒载体转染兔 BMSCs。采用甲苯胺蓝、番红 O 染色和实时 PCR 检查软骨分化情况。结果表明,Sox9 可有效诱导 BMSCs 在单层和 PGA 支架上的软骨生成。建立兔全层软骨缺损模型,然后用 PGA 支架和转导 Sox9 的兔 BMSCs 修复缺损。采用 HE、番红 O 染色和免疫组织化学方法评估复合物对缺损的修复情况。与未转导组相比,转导组的 PGA 支架和 BMSCs 修复效果更好,包括更多新形成的软骨组织和透明软骨特异性细胞外基质,以及几个软骨生成标志物基因的表达水平更高。本研究结果明确了 Sox9 在体内软骨缺损修复中的重要作用,并为 Sox9 在组织工程应用中的潜力和优势提供了证据。

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