Design and Evaluation of Dendrimer/Cyclodextrin Conjugate as a Novel Gene Carrier

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To design a novel gene transfer carrier, we prepared dendrimer conjugates with α-cyclodextrin (α-CDE conjugates) with its various degree of substitution (DS) and investigated the chemical structure, physicochemical properties, in vitro and in vivo gene transfer activity and cytotoxicity. α-CDE conjugates formed the complexes with plasmid DNA (pDNA), resulting in a change of the particle sizes of pDNA complexes, but the distinction of physicochemical properties among their vector/pDNA complexes was only very slight. The membrane-disruptive ability of α-CDE conjugates on liposomes encapsulating calcein and their cytotoxicity to NIH3T3 and HepG2 increased with an increase in the DS value of α-cyclodextrin. In vitro gene transfer activity of α-CDE conjugates in both NIH3T3 and HepG2 cells augmented as the charge ratio (vector/pDNA) increased, and the activity of α-CDE conjugate (G3, DS 2.4) was the highest at higher charge ratios among dendrimer (G3), the three α-CDE conjugates, and commercial transfection reagents such as Lipofectin and TransFast. After intravenous administration of pDNA complexes in mice, α-CDE conjugate (DS 2.4) delivered pDNA more efficiently in spleen, liver, and kidney, compared with dendrimer and other α-CDE conjugates. The potential use of α-CDE conjugate (G3, DS 2.4) could be expected as a nonviral vector in vitro and in vivo, and these data may be useful for design of α-CyD conjugates with other nonviral vectors. Furthermore, α-CDE conjugates attached with galactose (GalCDE conjugates) improved gene transfer activity more than parent α-CDE conjugates in HepG2 and NIH3T3 cells, suggesting that GalCDE conjugates are superior gene transfer carrier to the other commercially-available transfection agents.