RGD Peptide-Modified Dendrimer-Entrapped Gold Nanoparticles Enable Highly Efficient and Specific Gene Delivery to Stem Cells
RGD Peptide-Modified Dendrimer-Entrapped Gold Nanoparticles Enable Highly Efficient and Specific Gene Delivery to Stem Cells
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DOI：
10.1021/am508760w
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发表时间：
2015-03-04
影响因子：
9.5
通讯作者：
Shi, Xiangyang
中科院分区：
文献类型：
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作者：
Kong, Lingdan;Alves, Carla S.;Shi, Xiangyang
我们报道了使用精氨酸-甘氨酸-天冬氨酸(Arg-Gly-Asp，RGD)多肽修饰的树枝状大分子包裹的金纳米颗粒(Au DENPs)高效和特异地将基因输送到干细胞。本研究以聚乙二醇单甲醚和聚乙二醇单甲醚修饰RGD的第五代聚酰胺胺树枝状大分子为模板，制备了金纳米粒子。以天然和RGD修饰的聚乙二醇化树枝状大分子和性质良好的Au DENPs为载体，将携带增强型绿色荧光蛋白和荧光素酶(PEGFPLuc)报告基因的质粒DNA(PDNA)以及编码人骨形态发生蛋白-2(hBMP-2)基因的pDNA导入人骨髓间充质干细胞(HMSCs)。结果表明，所有载体均能同时导入两种pDNA的hMSCs。定量LUC活性检测和荧光显微镜定性评价证实了pEGFPLuc基因的导入。通过碱性磷酸酶活性、骨钙素分泌、钙沉积和von Kossa染色检测hBMP-2浓度和成骨分化程度，评价hBMP-2基因转导效率。我们的结果表明，干细胞基因的传递效率在很大程度上取决于树枝状大分子载体的组成和表面功能。RGD和AuNPs的共存使得所设计的树状大分子载体可能通过结合细胞表面的整合素受体和改善树枝状大分子的三维构象而具有与干细胞特异结合的能力，这有利于高效和特异的干细胞基因传递应用。
We report the use of arginine-glycine-aspartic (Arg-Gly-Asp, RGD) peptide-modified dendrimer-entrapped gold nanoparticles (Au DENPs) for highly efficient and specific gene delivery to stem cells. In this study, generation 5 poly(amidoamine) dendrimers modified with RGD via a poly(ethylene glycol) (PEG) spacer and with PEG monomethyl ether were used as templates to entrap gold nanoparticles (AuNPs). The native and the RGD-modified PEGylated dendrimers and the respective well characterized Au DENPs were used as vectors to transfect human mesenchymal stem cells (hMSCs) with plasmid DNA (pDNA) carrying both the enhanced green fluorescent protein and the luciferase (pEGFPLuc) reporter genes, as well as pDNA encoding the human bone morphogenetic protein-2 (hBMP-2) gene. We show that all vectors are capable of transfecting the hMSCs with both pDNAs. Gene transfection using pEGFPLuc was demonstrated by quantitative Luc activity assay and qualitative evaluation by fluorescence microscopy. For the transfection with hBMP-2, the gene delivery efficiency was evaluated by monitoring the hBMP-2 concentration and the level of osteogenic differentiation of the hMSCs via alkaline phosphatase activity, osteocalcin secretion, calcium deposition, and von Kossa staining assays. Our results reveal that the stem cell gene delivery efficiency is largely dependent on the composition and the surface functionality of the dendrimer-based vectors. The coexistence of RGD and AuNPs rendered the designed dendrimeric vector with specific stem cell binding ability likely via binding of integrin receptor on the cell surface and improved three-dimensional conformation of dendrimers, which is beneficial for highly efficient and specific stem cell gene delivery applications.