Efficient gene delivery to mesenchymal stem cells by an ethylenediamine-modified polysaccharide from mulberry leaves.
Efficient gene delivery to mesenchymal stem cells by an ethylenediamine-modified polysaccharide from mulberry leaves.
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DOI：
10.1002/smll.201101554
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发表时间：
2012-02
期刊：
影响因子：
13.3
通讯作者：
Wenwen Deng;Xia Cao;Miao Wang;Yan Yang;Weiyan Su;Yangyang Wei;Zhen Ou-yang;Jiangnan Yu;Ximing Xu
中科院分区：
文献类型：
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作者：
Wenwen Deng;Xia Cao;Miao Wang;Yan Yang;Weiyan Su;Yangyang Wei;Zhen Ou-yang;Jiangnan Yu;Ximing Xu
本研究探讨了利用桑葚中提取的天然多糖作为非病毒基因载体。将乙二胺化学接枝到桑葚多糖（MPS）的主链上，以获得核酸结合亲和力。粒径观察表明，阳离子桑葚多糖（CMPS）能有效地与质粒转化生长因子β1（TGF-β1）联合收割机结合形成纳米级颗粒。此外，电泳试验表明，当CMPS/pTGF-β1重量比增加至30：1时，质粒迁移延迟。基于来自大鼠股骨和胫骨的骨髓间充质干细胞（MSC）进行体外细胞转染实验，结果显示CMPS/pTGF-β1重量比为50：1的复合物表现出最高的细胞转染效果，其显著高于支链聚乙烯亚胺（PEI）（25 kDa; p = 0.001，Student's t-test）并且略高于Lipofectamine 2000。细胞毒性实验表明，所有复合物和质粒TGF-β1对骨髓间充质干细胞（MSCs）均无毒性。活细胞成像的结果证实，与阳性对照Lipofectamine 2000相比，MSC可以以更快的速率摄取更多的CMPS/质粒TGF-β1纳米颗粒;这些数据与转染效率数据一致。总之，这些结果表明CMPS/pTGF-β1纳米颗粒可以潜在地开发成用于将治疗性基因转移到细胞中的有前景的替代物。
This study investigates the use of a natural polysaccharide isolated from mulberry leaves as a nonviral gene vector. Ethylenediamine is chemically grafted to the backbone of a polysaccharide from mulberry leaves (MPS) to acquire nucleic acid binding affinity. A particle-size observation indicates that the cationic mulberry leaf polysaccharide (CMPS) can efficiently combine with plasmid transforming growth factor β1 (TGF-β1) to form nanoscaled particles. In addition, the electrophoresis assay indicates a retarded plasmid migration when the CMPS/pTGF-β1 weight ratio is increased to 30:1. The in vitro cell transfection experiment is performed based on bone marrow mesenchymal stem cells (MSCs) derived from rat femurs and tibias, and the findings reveal that the complex with a CMPS/pTGF-β1 weight ratio of 50:1 exhibits the highest cell transfection effect, which is significantly higher than that of branched poly(ethyleneimine) (PEI) (25 kDa; p = 0.001, Student's t-test) and slightly higher than Lipofectamine 2000. Moreover, the cytotoxicity assay also demonstrates that all of these tested complexes and the plasmid TGF-β1 are nontoxic to mesenchymal stem cells (MSCs). The results of the living cell imaging confirm that more of the CMPS/plasmid TGF-β1 nanoparticles can be taken up and at a faster rate by the MSCs than by the positive control Lipofectamine 2000; these data are consistent with the transfection efficiency data. Together, these results suggest that the CMPS/pTGF-β1 nanoparticle can potentially be developed into a promising alternative for the transfer of therapeutic genes into cells.