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聚酰胺-胺树枝状大分子功能化碳纳米管介导 GFP 基因转染 HeLa 细胞:不同类型碳纳米管的影响。

Polyamidoamine dendrimer-functionalized carbon nanotubes-mediated GFP gene transfection for HeLa cells: effects of different types of carbon nanotubes.

机构信息

Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, People's Republic of China.

出版信息

J Biomed Mater Res A. 2011 Nov;99(2):231-9. doi: 10.1002/jbm.a.33180. Epub 2011 Aug 16.

Abstract

Three types of functionalized carbon nanotubes (f-CNTs), polyamidoamine (PAMAM) dendrimer-functionalized single and multi-walled CNTs (MWCNT-PAMAM-1, MWCNT-PAMAM-2, and SWCNT-PAMAM-3), were prepared by covalent linkage approach. The micro-morphologies of the three f-CNTs and the interaction of MWCNT-PAMAM-2 with HeLa cells were characterized by transmission electron microscopy (TEM). The free amine groups on the surface of the three types of CNTs-PAMAM hybrids were quantitatively analyzed. Their cytotoxicity and transfection efficiency of plasmid DNA of enhanced green fluorescent protein (pEGFP-N1) to HeLa cells were investigated in detail. The results suggest that although all three types of CNTs-PAMAM hybrids can deliver pEGFP-N1 into HeLa cells and the exogenous GFP gene has been successfully expressed, MWCNT-PAMAM-2 with shorter length and larger amount of free amine groups on its surface possesses higher transfection efficiency (6.79%), being about 3.0 and 1.7 times as large as those of MWCNT-PAMAM-1 (2.24%) and SWCNT-PAMAM-3 (4.08%), respectively; their cytotoxicity to HeLa cells decrease following the sequence of SWCNT-PAMAM-3 > MWCNT-PAMAM-2 > MWCNT-PAMAM-1. These results may be useful for understanding the effects of the chemical/physical properties of f-CNTs on their gene transfection efficiency and cytotoxicity, allowing for construction of promising CNT-based intracellular delivery vectors for gene therapy.

摘要

三种功能化碳纳米管(f-CNTs),即聚酰胺-胺(PAMAM)树枝状大分子功能化的单壁和多壁 CNT(MWCNT-PAMAM-1、MWCNT-PAMAM-2 和 SWCNT-PAMAM-3),通过共价键合方法制备。通过透射电子显微镜(TEM)对三种 f-CNTs 的微观形貌和 MWCNT-PAMAM-2 与 HeLa 细胞的相互作用进行了表征。定量分析了三种 CNTs-PAMAM 杂化物表面的游离胺基。详细研究了它们对 HeLa 细胞的细胞毒性和质粒 DNA 增强型绿色荧光蛋白(pEGFP-N1)的转染效率。结果表明,尽管三种 CNTs-PAMAM 杂化物都可以将 pEGFP-N1 递送至 HeLa 细胞,并且外源 GFP 基因已成功表达,但表面具有较短长度和更多游离胺基的 MWCNT-PAMAM-2 具有更高的转染效率(6.79%),分别约为 MWCNT-PAMAM-1(2.24%)和 SWCNT-PAMAM-3(4.08%)的 3.0 倍和 1.7 倍;它们对 HeLa 细胞的细胞毒性按 SWCNT-PAMAM-3 > MWCNT-PAMAM-2 > MWCNT-PAMAM-1 的顺序降低。这些结果可能有助于理解 f-CNTs 的化学/物理性质对其基因转染效率和细胞毒性的影响,从而构建有前途的基于 CNT 的细胞内递药载体用于基因治疗。

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