Efficient Dendrimer鈥揇NA Complexation and Gene Delivery Vector Properties of Nitrogen-Core Poly(propyl ether imine) Dendrimer in Mammalian Cells

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• 作者：Abirami Lakshminarayanan ; Vijay Kumar Ravi ; Ranjitha Tatineni ; Y. B. R. D. Rajesh ; Vishal Maingi ; K. S. Vasu ; Nandhitha Madhusudhan ; Prabal K. Maiti ; A. K. Sood ; Saumitra Das ; N. Jayaraman
• 刊名：Bioconjugate Chemistry
• 出版年：2013
• 出版时间：September 18, 2013
• 年：2013
• 卷：24
• 期：9
• 页码：1612-1623
• 全文大小：628K
• 年卷期：v.24,no.9(September 18, 2013)
• ISSN：1520-4812

文摘

Dendrimers as vectors for gene delivery were established, primarily by utilizing few prominent dendrimer types so far. We report herein studies of DNA complexation efficacies and gene delivery vector properties of a nitrogen-core poly(propyl ether imine) (PETIM) dendrimer, constituted with 22 tertiary amine internal branches and 24 primary amines at the periphery. The interaction of the dendrimer with pEGFPDNA was evaluated through UV鈥搗is, circular dichroism (CD) spectral studies, ethidium bromide fluorescence emission quenching, thermal melting, and gel retardation assays, from which most changes to DNA structure during complexation was found to occur at a weight ratio of dendrimer:DNA 2:1. The zeta potential measurements further confirmed this stoichiometry at electroneutrality. The structure of a DNA oligomer upon dendrimer complexation was simulated through molecular modeling and the simulation showed that the dendrimer enfolded DNA oligomer along both major and minor grooves, without causing DNA deformation, in 1:1 and 2:1 dendrimer-to-DNA complexes. Atomic force microscopy (AFM) studies on dendrimer-pEGFP DNA complex showed an increase in the average z-height as a result of dendrimers decorating the DNA, without causing a distortion of the DNA structure. Cytotoxicity studies involving five different mammalian cell lines, using [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) assay, reveal the dendrimer toxicity profile (IC₅₀) values of 400鈥?000 渭g mL^鈥?, depending on the cell line tested. Quantitative estimation, using luciferase assay, showed that the gene transfection was at least 100 times higher when compared to poly(ethylene imine) branched polymer, having similar number of cationic sites as the dendrimer. The present study establishes the physicochemical behavior of new nitrogen-core PETIM dendrimer鈥揇NA complexes, their lower toxicities, and efficient gene delivery vector properties.