A doxycycline inducible, adenoviral bone morphogenetic protein-2 gene delivery system to bone
A doxycycline inducible, adenoviral bone morphogenetic protein-2 gene delivery system to bone
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DOI：
10.1002/term.2393
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发表时间：
2018-01-01
影响因子：
3.3
通讯作者：
Stoddart, Martin J.
中科院分区：
文献类型：
--
作者：
Bara, Jennifer J.;Dresing, Iska;Stoddart, Martin J.
我们报告了一种可调的、非整合的病毒基因骨传递系统的新用途，该系统可以以“现成”的方式与临床批准的生物材料结合。具体地，使用多西环素诱导的Tet-on腺病毒载体（AdTetBMP-2）与间充质基质细胞（MSC）、纤维蛋白和双相磷酸钙陶瓷（MBCP（R））的组合来修复裸大鼠中的大的骨缺损。骨形态发生蛋白-2（BMP-2）转基因表达可以通过修改强力霉素浓度进行有效调节。腺病毒BMP-2基因递送对骨愈合的影响在体内在4 mm临界尺寸的、内固定的股骨缺损中进行了研究。通过直接应用AdTetBMP-2或通过用病毒预包被MBCP颗粒来转导MSC。在递送AdTetBMP-2后，死后的放射学评估评分显著改善。在AdTetBMP-2组中，组织学分析显示，与对照组相比，缺损部位有更多的新骨形成。新形成的骨血管化，并与新生组织和植入的生物材料完全整合。使用两种载体递送方法（直接施用与预涂覆MCBP）实现了愈合结果的改善。腺病毒介导的BMP-2增强了骨髓间充质干细胞、纤维蛋白和MBCP体内移植所实现的骨再生。重要的是，我们的体外和体内数据表明，这可以用相对低（ng/ml）水平的生长因子实现。我们的模型和新的基因传递系统可能提供一个强大的标准化工具，优化生长因子的交付和释放，愈合大骨缺损。版权所有（C）2016约翰威利父子有限公司
We report the novel use of a tuneable, non-integrating viral gene delivery system to bone that can be combined with clinically approved biomaterials in an 'off-the-shelf' manner. Specifically, a doxycycline inducible Tet-on adenoviral vector (AdTetBMP-2) in combination with mesenchymal stromal cells (MSCs), fibrin and a biphasic calcium phosphate ceramic (MBCP (R)) was used to repair large bone defects in nude rats. Bone morphogenetic protein-2 (BMP-2) transgene expression could be effectively tuned by modification of the doxycycline concentration. The effect of adenoviral BMP-2 gene delivery upon bone healing was investigated in vivo in 4 mm critically sized, internally fixated, femoral defects. MSCs were transduced either by direct application of AdTetBMP-2 or by pre-coating MBCP granules with the virus. Radiological assessment scores post-mortem were significantly improved upon delivery of AdTetBMP-2. In AdTetBMP-2 groups, histological analysis revealed significantly more newly formed bone at the defect site compared with controls. Newly formed bone was vascularized and fully integrated with nascent tissue and implanted biomaterial. Improvement in healing outcome was achieved using both methods of vector delivery (direct application vs. pre-coating MCBP). Adenoviral delivery of BMP-2 enhanced bone regeneration achieved by the transplantation of MSCs, fibrin and MBCP in vivo. Importantly, our in vitro and in vivo data suggest that this can be achieved with relatively low (ng/ml) levels of the growth factor. Our model and novel gene delivery system may provide a powerful standardized tool for the optimization of growth factor delivery and release for the healing of large bone defects. Copyright (C) 2016 John Wiley & Sons, Ltd.