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Biomacromolecules ; 12(6): 2243-50, 2011 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-21557603

RESUMO

Conventional free radical polymerization with subsequent postpolymerization modification afforded imidazolium copolymers with controlled charge density and side chain hydroxyl number. Novel imidazolium-containing copolymers where each permanent cation contained one or two adjacent hydroxyls allowed precise structure-transfection efficiency studies. The degree of polymerization was identical for all copolymers to eliminate the influence of molecular weight on transfection efficiency. DNA binding, cytotoxicity, and in vitro gene transfection in African green monkey COS-7 cells revealed structure-property-transfection relationships for the copolymers. DNA gel shift assays indicated that higher charge densities and hydroxyl concentrations increased DNA binding. As the charge density of the copolymers increased, toxicity of the copolymers also increased; however, as hydroxyl concentration increased, cytotoxicity remained constant. Changing both charge density and hydroxyl levels in a systematic fashion revealed a dramatic influence on transfection efficiency. Dynamic light scattering of the polyplexes, which were composed of copolymer concentrations required for the highest luciferase expression, showed an intermediate DNA-copolymer binding affinity. Our studies supported the conclusion that cationic copolymer binding affinity significantly impacts overall transfection efficiency of DNA delivery vehicles, and the incorporation of hydroxyl sites offers a less toxic and effective alternative to more conventional highly charged copolymers.


Assuntos
DNA/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Técnicas de Transferência de Genes , Hidróxidos/química , Imidazóis/síntese química , Luciferases/metabolismo , Plasmídeos/metabolismo , Polímeros/síntese química , Animais , Células COS , Cátions/química , Cátions/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , DNA/genética , DNA/farmacologia , Ensaio de Desvio de Mobilidade Eletroforética , Terapia Genética/métodos , Ligação de Hidrogênio , Hidróxidos/metabolismo , Imidazóis/metabolismo , Luciferases/genética , Microscopia de Fluorescência , Plasmídeos/genética , Plasmídeos/farmacologia , Polímeros/metabolismo , Eletricidade Estática , Transfecção
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