Department of General Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
Colloids Surf B Biointerfaces. 2013 Sep 1;109:183-9. doi: 10.1016/j.colsurfb.2013.03.045. Epub 2013 Apr 8.
Treatment of HIV infection by gene therapy is a promising tool for combating AIDS. One of the primary limitations of gene therapy is the effective delivery of nucleic acids to the target cells. Dendrimers are nanoparticles that are increasingly being used as nucleic acid vehicles. We have synthesized "Si-C" amino-terminated carbosilane dendrimers GnO3(NMe3)m functionalized with quaternary ammonium (NMe3(+)) terminal groups via hydrosilylation of allyl dimethylamine with the corresponding GnO3(SiH)m dendrimers and further addition of MeI. These dendrimers are soluble in water. Initially, complexation between these "Si-C" dendrimers and anti-HIV nucleic acids (oligodeoxynucleotides ANTITAR and GEM91, siRNA siP24) was studied and molar ratios for complete complexation were determined. Then the charge and size of the dendriplexes (complexes of "Si-C" dendrimers with nucleic acids) were analyzed and it was found that they possessed charges of +5 to +40 mV and sizes of 60-600 nm (zeta-size) or 50-100 nm (atomic force microscopy) suitable for cell transfection. Stability studies showed that the dendriplexes were stable over time and were resistant to degradation by serum albumin. The effects of dendrimers and their dendriplexes on erythrocytes (isolated and in whole blood) revealed that the dendriplexes were significantly less cytotoxic than the pure dendrimers. The effects of dendrimers and their dendriplexes on peripheral blood mononuclear cells (the main target of HIV) were analyzed and it was found that the dendriplexes were 10 times less cytotoxic than the pure dendrimers. Finally, transfection experiments revealed that "Si-C"-carbosilane dendrimers had a restricted ability to deliver long-chain double-stranded nucleic acids. The results indicate that these cationic carbosilane dendrimers are good candidates for delivering short-chain siRNA and oligodeoxynucleotide to HIV-infected peripheral blood mononuclear cells or lymphocytes.
基因治疗治疗 HIV 感染是一种对抗艾滋病的有前途的工具。基因治疗的主要限制之一是有效将核酸递送到靶细胞。树突状聚合物是越来越多地用作核酸载体的纳米颗粒。我们已经通过烯丙基二甲基胺与相应的 GnO3(SiH)m 树突状聚合物的氢硅烷化反应以及进一步添加 MeI 来合成了“Si-C”末端氨基封端的碳硅烷树突GnO3(NMe3)m。这些树突状聚合物可溶于水。最初，研究了这些“Si-C”树突状聚合物与抗 HIV 核酸（寡脱氧核苷酸 ANTITAR 和 GEM91、siRNA siP24）之间的络合，并确定了完全络合的摩尔比。然后分析了树突状聚合物（“Si-C”树突状聚合物与核酸的复合物）的电荷和大小，发现它们具有 +5 至 +40 mV 的电荷和 60-600nm（ζ-尺寸）或 50-100nm（原子力显微镜）的尺寸，适合细胞转染。稳定性研究表明，树突状聚合物随时间稳定并且能够抵抗血清白蛋白的降解。树突状聚合物及其树突状聚合物对红细胞（分离和全血）的影响表明，树突状聚合物的细胞毒性明显低于纯树突状聚合物。分析了树突状聚合物及其树突状聚合物对外周血单个核细胞（HIV 的主要靶标）的影响，发现树突状聚合物的细胞毒性比纯树突状聚合物低 10 倍。最后，转染实验表明，“Si-C”-碳硅烷树突状聚合物递送长链双链核酸的能力有限。结果表明，这些阳离子碳硅烷树突状聚合物是将短链 siRNA 和寡脱氧核苷酸递送到 HIV 感染的外周血单个核细胞或淋巴细胞的良好候选物。
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