title: Efficient dendrimer-DNA complexation and gene delivery vector properties of nitrogen-core poly(propyl ether imine) dendrimer in mammalian cells.
abstract: 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-vis, 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 ∼400-1000 μg mL(-1), 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-DNA complexes, their lower toxicities, and efficient gene delivery vector properties.
authors.affiliation: Department of Organic Chemistry, ‡Department of Physics, and §Department of Microbiology and Cell Biology, Indian Institute of Science , Bangalore 560 012, India.
authors.forename: Abirami
authors.initials: A
authors.lastname: Lakshminarayanan
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Ravi
authors.forename: Ranjitha
authors.initials: R
authors.lastname: Tatineni
authors.forename: Y B R D
authors.initials: YB
authors.lastname: Rajesh
authors.forename: Vishal
authors.initials: V
authors.lastname: Maingi
authors.forename: K S
authors.initials: KS
authors.lastname: Vasu
authors.forename: Nandhitha
authors.initials: N
authors.lastname: Madhusudhan
authors.forename: Prabal K
authors.initials: PK
authors.lastname: Maiti
authors.forename: A K
authors.initials: AK
authors.lastname: Sood
authors.forename: Saumitra
authors.initials: S
authors.lastname: Das
authors.forename: N
authors.initials: N
authors.lastname: Jayaraman
mesh_terms: D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D050091:Dendrimers / Q000737:chemistry* / Q000633:toxicity; D004987:Ethers / Q000737:chemistry* / Q000633:toxicity; D006801:Humans; D007097:Imines / Q000737:chemistry* / Q000633:toxicity; D056004:Molecular Dynamics Simulation; D014162:Transfection
chemical_list: D050091:Dendrimers; D004987:Ethers; D007097:Imines; C045122:propyl ether; D004247:DNA