New scaffolds encapsulating TGF-β3/BMP-7 combinations driving strong chondrogenic differentiation
New scaffolds encapsulating TGF-β3/BMP-7 combinations driving strong chondrogenic differentiation
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DOI：
10.1016/j.ejpb.2016.12.021
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发表时间：
2017-05-01
影响因子：
4.9
通讯作者：
Garcia-Fuentes, Marcos
中科院分区：
文献类型：
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作者：
Crecente-Campo, Jose;Borrajo, Erea;Garcia-Fuentes, Marcos
尽管目前可以获得多种用与该应用相关的生长因子激活的组织支架，但关节软骨的再生仍然是一个未解决的问题。将这些因素中的多个因素结合起来可能会带来进一步的进步；在这里，我们提出了一种针对 BMP-7 和 TGF-β 3 这两种具有软骨形成活性的蛋白质的双重递送而优化的支架组合物。首先，我们在含有 TGF-β 3 的间充质干细胞微团培养物中测试暴露于负载 BMP-7 的微球是否会改善软骨形成。组织学和 qRT-PCR 数据证实，BMP-7 的持续释放与 TGF-β 3 协同作用，促进软骨分化。然后，我们优化了用于组织培养和 BMP-7 和 TGF-β 3 双重封装的支架原型。支架由聚乳酸-乙醇酸共聚物制备，BMP-7/TGF-β 3 与肝素和 Tetronic 1107 一起作为纳米复合物负载。支架显示两种蛋白在 4 周内持续释放， 最小的爆发效应。我们最终在没有外源软骨形成因子补充的情况下在这些支架上培养人类间充质干细胞。在负载有BMP-7和TGF-β3的支架上培养的细胞在宏观和组织学上显示出明显的软骨形成迹象。 RT-PCR 研究证实软骨标记物 SOX9 和聚集蛋白聚糖明显上调。总之，封装 BMP-7 和 TGF-β 3 的支架可以有效地递送协同生长因子组合，从而驱动人间充质干细胞培养物中有效的软骨形成。这些结果为该技术在软骨再生实验中的体内转化开辟了有吸引力的前景。 (C) 2017 Elsevier B.V. 保留所有权利。
The regeneration of articular cartilage remains an unresolved question despite the current access to a variety of tissue scaffolds activated with growth factors relevant to this application. Further advances might result from combining more than one of these factors; here, we propose a scaffold composition optimized for the dual delivery of BMP-7 and TGF-beta 3, two proteins with described chondrogenic activity. First, we tested in a mesenchymal stem cell micromass culture with TGF-beta 3 whether the exposure to microspheres loaded with BMP-7 would improve cartilage formation. Histology and qRT-PCR data confirmed that the sustained release of BMP-7 cooperates with TGF-beta 3 towards chondrogenic differentiation. Then, we optimized a scaffold prototype for tissue culture and dual encapsulation of BMP-7 and TGF-beta 3. The scaffolds were prepared from poly(lactic-co-glycolic acid), and BMP-7/TGF-beta 3 were loaded as nanocomplexes with heparin and Tetronic 1107. The scaffolds showed the sustained release of both proteins over four weeks, with minimal burst effect. We finally cultured human mesenchymal stem cells on these scaffolds, in the absence of exogenous chondrogenic factor supplementation. The cells cultured on the scaffolds loaded with BMP-7 and TGF-beta 3 showed clear signs of cartilage formation macroscopically and histologically. RT-PCR studies confirmed a clear upregulation of cartilage markers SOX9 and Aggrecan. In summary, scaffolds encapsulating BMP-7 and TGF-beta 3 can efficiently deliver a cooperative growth factor combination that drives efficient cartilage formation in human mesenchymal stem cell cultures. These results open attractive perspectives towards in vivo translation of this technology in cartilage regeneration experiments. (C) 2017 Elsevier B.V. All rights reserved.