Repair of full-thickness articular cartilage defects by cultured mesenchymal stem cells transfected with the transforming growth factor β1 gene
Repair of full-thickness articular cartilage defects by cultured mesenchymal stem cells transfected with the transforming growth factor β1 gene
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DOI：
10.1088/1748-6041/1/4/006
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发表时间：
2006-12-01
影响因子：
4
通讯作者：
Quan, Daping
中科院分区：
文献类型：
--
作者：
Guo, Xiaodong;Zheng, Qixin;Quan, Daping
关节软骨修复仍然是一个临床和科学的挑战，越来越多的兴趣集中在基因转移和组织工程的组合技术。转化生长因子β 1（TGF-β 1）是一种多功能分子，在促进软骨修复、抑制炎症和同种异体反应性免疫反应中发挥核心作用。细胞介导的基因治疗可以允许TGF-β（1）的持续表达，这可能会避免与生长因子传递相关的困难。本研究旨在探讨TGF-β 1基因修饰的间充质干细胞（MSCs）是否能促进同种异体兔关节软骨全层缺损的修复。将转染pcDNA（3）-TGF-β（1）基因的骨髓间充质干细胞接种于可生物降解的聚L-赖氨酸包被的聚乳酸（PLA）仿生支架上，并移植于18只新西兰兔关节软骨全层缺损处。以转染pcDNA 3基因的MSCs/仿生支架复合物和无细胞支架分别作为对照组I和对照组II。随访时间分别为2、4、12和24周。进行了宏观、组织学和超微结构研究。体外扫描电镜研究发现，实验组在接种后两周产生了丰富的软骨基质，并完全覆盖了支架的互连孔。在体内，随着时间的推移，再生组织的质量得到改善，在植入后24周，透明软骨填充软骨区域，小梁和密质骨的混合物填充软骨下区域。实验组的关节修复优于对照组I或II，在以下方面：（1）在缺损上部合成透明软骨特异性细胞外基质;（2）在缺损下部重建软骨下骨;（3）抑制炎症和同种异体免疫反应。转染的MSC在体内过表达其TGF-β（1）基因产物至少4周。对照缺损填充纤维和纤维软骨组织的混合物。本研究中使用的TGF-β 1基因转染的MSCs/聚-L-赖氨酸包被的PLA复合同种异体移植物对关节软骨修复是有效的。这种新的TGF-β（1）基因增强的组织工程策略可能对增强受损关节软骨的修复，特别是由退行性疾病引起的这种损伤具有潜在的益处。
Articular cartilage repair remains a clinical and scientific challenge with increasing interest focused on the combined techniques of gene transfer and tissue engineering. Transforming growth factor beta 1 (TGF-beta(1)) is a multifunctional molecule that plays a central role in promotion of cartilage repair, and inhibition of inflammatory and alloreactive immune response. Cell mediated gene therapy can allow a sustained expression of TGF-beta(1) that may circumvent difficulties associated with growth factor delivery. The objective of this study was to investigate whether TGF-beta(1) gene modified mesenchymal stem cells (MSCs) could enhance the repair of full-thickness articular cartilage defects in allogeneic rabbits. The pcDNA(3)-TGF-beta(1) gene transfected MSCs were seeded onto biodegradable poly-L-lysine coated polylactide (PLA) biomimetic scaffolds in vitro and allografted into full-thickness articular cartilage defects in 18 New Zealand rabbits. The pcDNA(3) gene transfected MSCs/biomimetic scaffold composites and the cell-free scaffolds were taken as control groups I and II,respectively. The follow-up times were 2, 4, 12 and 24 weeks. Macroscopical, histological and ultrastructural studies were performed. In vitro SEM studies found that abundant cartilaginous matrices were generated and completely covered the interconnected pores of the scaffolds two weeks post-seeding in the experimental groups. In vivo, the quality of regenerated tissue improved over time with hyaline cartilage filling the chondral region and a mixture of trabecular and compact bone filling the subchondral region at 24 weeks post-implantation. Joint repair in the experimental groups was better than that of either control group I or II, with respect to: ( 1) synthesis of hyaline cartilage specific extracellular matrix at the upper portion of the defect; ( 2) reconstitution of the subchondral bone at the lower portion of the defect and ( 3) inhibition of inflammatory and alloreactive immune responses. The transfected MSCs overexpressed their TGF-beta(1) gene products for at least 4 weeks in vivo. The control defects were filled with a mixture of fibrous and fibrocartilaginous tissue. The TGF-beta(1) gene transfected MSCs/poly-L-lysine coated PLA composite allografts used in this study are effective for articular cartilage repair. This novel TGF-beta(1) gene enhanced tissue engineering strategy may be of potential benefit to enhancing the repair of damaged articular cartilage, especially such damage caused by degenerative disease.