title: Pentablock copolymers of pluronic F127 and modified poly(2-dimethyl amino)ethyl methacrylate for internalization mechanism and gene transfection studies.
abstract: Cationic polymers are one of the major nonviral gene delivery vectors investigated in the past decade. In this study, we synthesized several cationic copolymers using atom transfer radical polymerization (ATRP) for gene delivery vectors: pluronic F127-poly(dimethylaminoethyl methacrylate) (PF127-pDMAEMA), pluronic F127-poly (dimethylaminoethyl methacrylate-tert-butyl acrylate) (PF127-p(DMAEMA-tBA)), and pluronic F127-poly(dimethylaminoethyl methacrylate-acrylic acid) (PF127-p(DMAEMA-AA)). The copolymers showed high buffering capacity and efficiently complexed with plasmid deoxyribonucleic acid (pDNA) to form nanoparticles 80-180 nm in diameter and with positive zeta potentials. In the absence of 10% fetal bovine serum, PF127-p(DMAEMA-AA) showed the highest gene expression and the lowest cytotoxicity in 293T cells. After acrylic acid groups had been linked with a fluorescent dye, the confocal laser scanning microscopic image showed that PF127-p(DMAEMA-AA)/pDNA could efficiently enter the cells. Both clathrin-mediated and caveolae-mediated endocytosis mechanisms were involved. Our results showed that PF127-p(DMAEMA-AA) has great potential to be a gene delivery vector.
authors.affiliation: Department of Medicinal and Applied Chemistry, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Shih-Jer
authors.initials: SJ
authors.lastname: Huang
authors.forename: Tzu-Pin
authors.initials: TP
authors.lastname: Wang
authors.forename: Sheng-I
authors.initials: SI
authors.lastname: Lue
authors.forename: Li-Fang
authors.initials: LF
authors.lastname: Wang
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D008689:Methacrylates / Q000737:chemistry*; D008970:Molecular Weight; D009757:Nylons / Q000737:chemistry*; D020442:Poloxamer / Q000737:chemistry*; D058105:Polymerization; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity; D013057:Spectrum Analysis; D014162:Transfection / Q000379:methods*
chemical_list: D008689:Methacrylates; D009757:Nylons; D011108:Polymers; C407037:poly(2-(dimethylamino)ethyl methacrylate); D020442:Poloxamer
keywords: atom transfer radical polymerization; copolymer; dimethylaminoethyl methacrylate; nonviral vector; pluronic F127