Nanoplex-Mediated Codelivery of Fibroblast Growth Factor and Bone Morphogenetic Protein Genes Promotes Osteogenesis in Human Adipocyte-Derived Mesenchymal Stem Cells.
Nanoplex-Mediated Codelivery of Fibroblast Growth Factor and Bone Morphogenetic Protein Genes Promotes Osteogenesis in Human Adipocyte-Derived Mesenchymal Stem Cells.
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DOI：
10.1021/acs.molpharmaceut.5b00297
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发表时间：
2015-08-03
影响因子：
4.9
通讯作者：
Salem AK
中科院分区：
文献类型：
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作者：
Atluri K;Seabold D;Hong L;Elangovan S;Salem AK
这项研究强调了转染介导的协调骨形态发生蛋白2（BMP-2）和成纤维细胞生长因子2（FGF-2）信号在促进成骨中的重要性。我们采用单独编码BMP-2和FGF-2的质粒与聚乙烯亚胺（PEI）复合，体外转染人脂肪间充质干细胞（hADMSCs）。对纳米复合物的大小、表面电荷、体外细胞毒性和在hADMSC中的转染能力进行表征。与用单独的pFGF-2或单独的pBMP-2的PEI纳米复合物转染相比，在用负载有pFGF-2和pBMP-2两者的PEI纳米复合物（PEI/（pFGF-2 + pBMP-2））转染hADMSC后第7天观察到BMP-2蛋白分泌的显著增强。通过使用真实的时间聚合酶链反应测量骨钙素和Runx-2基因表达来确定转染的hADMSCs的成骨分化。与用仅含有pFGF-2或pBMP-2的纳米复合物转染的细胞相比，用PEI/（pFGF-2 + pBMP-2）转染的细胞分别在转染后第3天和第7天观察到Runx-2和骨钙素表达的显著增加。茜素红染色和原子吸收光谱显示，与其他处理相比，在用PEI/（pFGF-2 + pBMP-2）转染后第14天和第30天，hADMSC培养物中钙沉积水平升高。我们已经表明，pFGF-2和pBMP-2的共同递送导致成骨蛋白合成、成骨标志物表达和随后的矿化的显著增强。这项研究指出了一种新的临床翻译策略，以实现有效的骨再生。
This study highlights the importance of transfection mediated coordinated bone morphogenetic protein 2 (BMP-2) and fibroblast growth factor 2 (FGF-2) signaling in promoting osteogenesis. We employed plasmids independently encoding BMP-2 and FGF-2 complexed with polyethylenimine (PEI) to transfect human adipose derived mesenchymal stem cells (hADMSCs) in vitro. The nanoplexes were characterized for size, surface charge, in vitro cytotoxicity and transfection ability in hADMSCs. A significant enhancement in BMP-2 protein secretion was observed on day 7 post-transfection of hADMSCs with PEI nanoplexes loaded with both pFGF-2 and pBMP-2 (PEI/(pFGF-2 + pBMP-2)) versus transfection with PEI nanoplexes of either pFGF-2 alone or pBMP-2 alone. Osteogenic differentiation of transfected hADMSCs was determined by measuring osteocalcin and Runx-2 gene expression using real time polymerase chain reactions. A significant increase in the expression of Runx-2 and osteocalcin was observed on day 3 and day 7 post-transfection, respectively, by cells transfected with PEI/(pFGF-2 + pBMP-2) compared to cells transfected with nanoplexes containing pFGF-2 or pBMP-2 alone. Alizarin Red staining and atomic absorption spectroscopy revealed elevated levels of calcium deposition in hADMSC cultures on day 14 and day 30 post-transfection with PEI/(pFGF-2 + pBMP-2) compared to other treatments. We have shown that co-delivery of pFGF-2 and pBMP-2 results in a significant enhancement in osteogenic protein synthesis, osteogenic marker expression and subsequent mineralization. This research points to a new clinically translatable strategy for achieving efficient bone regeneration.