Encapsulation of plasmid DNA in calcium phosphate nanoparticles: stem cell uptake and gene transfer efficiency.
Encapsulation of plasmid DNA in calcium phosphate nanoparticles: stem cell uptake and gene transfer efficiency.
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磷酸钙纳米颗粒中质粒 DNA 的封装：干细胞摄取和基因转移效率
DOI：
10.2147/ijn.s27370
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发表时间：
2011
影响因子：
8
通讯作者：
Xu X
中科院分区：
文献类型：
--
作者：
Cao X;Deng W;Wei Y;Su W;Yang Y;Wei Y;Yu J;Xu X
背景本研究的目的是开发磷酸钙纳米复合颗粒包裹质粒DNA（CP-pDNA）纳米颗粒作为非病毒载体的基因传递。方法制备转化生长因子β 1（TGF-β1）介导的CP-pDNA纳米粒，并对其进行表征。在间充质干细胞中评价CP-pDNA纳米颗粒的转染效率和细胞活力，通过免疫荧光染色鉴定。MTT法检测TGF-β1质粒和磷酸钙对骨髓间充质干细胞的细胞毒性。结果CP-pDNA纳米粒保持了TGF-β1的完整性。良好分散的CP-pDNA纳米颗粒表现出超低粒度（20-50 nm）和比Lipofectamine™ 2000显著更低的细胞毒性。免疫荧光染色显示，本实验培养的细胞可能为间充质干细胞。CP-pDNA纳米颗粒对间充质干细胞的细胞摄取和转染效率高于针状磷酸钙纳米颗粒和标准磷酸钙转染试剂盒。此外，活细胞成像和共聚焦激光显微镜生动地展示了CP-pDNA纳米颗粒在间充质干细胞中的运输过程。细胞毒性试验结果表明，TGF-β1质粒和磷酸钙对间充质干细胞无毒性。结论CP-pDNA纳米粒是一种高效、低毒的非病毒基因载体。
Background The purpose of this study was to develop calcium phosphate nanocomposite particles encapsulating plasmid DNA (CP-pDNA) nanoparticles as a nonviral vector for gene delivery. Methods CP-pDNA nanoparticles employing plasmid transforming growth factor beta 1 (TGF-β1) were prepared and characterized. The transfection efficiency and cell viability of the CP-pDNA nanoparticles were evaluated in mesenchymal stem cells, which were identified by immunofluorescence staining. Cytotoxicity of plasmid TGF-β1 and calcium phosphate to mesenchymal stem cells were evaluated by MTT assay. Results The integrity of TGF-β1 encapsulated in the CP-pDNA nanoparticles was maintained. The well dispersed CP-pDNA nanoparticles exhibited an ultralow particle size (20–50 nm) and significantly lower cytotoxicity than Lipofectamine™ 2000. Immunofluorescence staining revealed that the cultured cells in this study were probably mesenchymal stem cells. The cellular uptake and transfection efficiency of the CP-pDNA nanoparticles into the mesenchymal stem cells were higher than that of needle-like calcium phosphate nanoparticles and a standard calcium phosphate transfection kit. Furthermore, live cell imaging and confocal laser microscopy vividly showed the transportation process of the CP-pDNA nanoparticles in mesenchymal stem cells. The results of a cytotoxicity assay found that both plasmid TGF-β1 and calcium phosphate were not toxic to mesenchymal stem cells. Conclusion CP-pDNA nanoparticles can be developed into an effective alternative as a nonviral gene delivery system that is highly efficient and has low cytotoxicity.