title: Characterization of tailor-made copolymers of oligo(ethylene glycol) methyl ether methacrylate and N,N-dimethylaminoethyl methacrylate as nonviral gene transfer agents: influence of macromolecular structure on gene vector particle properties and transfection efficiency.
abstract: Oligo(ethylene glycol) methyl ether methacrylates (OEGMA) of various chain lengths (i.e., 9, 23, or 45 EG units) and N,N-dimethylaminoethyl methacrylate (DMAEMA) were copolymerized by atom transfer radical polymerization (ATRP), yielding well-defined P(DMAEMA-co-OEGMA) copolymers with increasing OEGMA molar fractions (F(OEGMA)) but a comparable degree of polymerization (DP approximately 120). Increase of both F(OEGMA) and OEGMA chain lengths correlated inversely with gene vector size, morphology, and zeta potential. P(DMAEMA-co-OEGMA) copolymers prevented gene vector aggregation at high plasmid DNA (pDNA) concentrations in isotonic solution and did not induce cytotoxicity even at high concentrations. Transfection efficiency of the most efficient P(DMAEMA-co-OEGMA) copolymers was found to be >10-fold lower compared with branched polyethylenimine (PEI) 25 kDa. Although OEGMA copolymerization largely reduced gene vector binding with the cell surface, cellular internalization of the bound complexes was less affected. These observations suggest that inefficient endolysosomal escape limits transfection efficiency of P(DMAEMA-co-OEGMA) copolymer gene vectors. Despite this observation, optimized p(DMAEMA-co-OEGMA) gene vectors remained stable under conditions for in vivo application leading to 7-fold greater gene expression in the lungs compared with PEI. Tailor-made P(DMAEMA-co-OEGMA) copolymers are promising nonviral gene transfer agents that fulfill the requirements for successful in vivo gene delivery.
authors: Department of Pediatrics, Ludwig-Maximilians University, 80337 Munich, Germany.
authors: Senta
authors: S
authors: Uzgün
authors: Ozgür
authors: O
authors: Akdemir
authors: Günther
authors: G
authors: Hasenpusch
authors: Christof
authors: C
authors: Maucksch
authors: Monika M
authors: MM
authors: Golas
authors: Bjoern
authors: B
authors: Sander
authors: Holger
authors: H
authors: Stark
authors: Rabea
authors: R
authors: Imker
authors: Jean-François
authors: JF
authors: Lutz
authors: Carsten
authors: C
authors: Rudolph
mesh_terms: D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002470:Cell Survival; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000494:pharmacology*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008689:Methacrylates / Q000737:chemistry*; D020366:Methylmethacrylate / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D011094:Polyethyleneimine; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism; D014162:Transfection
chemical_list: D004337:Drug Carriers; D008689:Methacrylates; D011108:Polymers; D020366:Methylmethacrylate; D011094:Polyethyleneimine; D004247:DNA; D008156:Luciferases; C049840:2-(dimethylamino)ethyl methacrylate