title: Biodegradable poly(vinyl alcohol)-polyethylenimine nanocomposites for enhanced gene expression in vitro and in vivo.
abstract: Use of cationic polymers as nonviral gene vectors has several limitations such as low transfection efficiency, high toxicity, and inactivation by serum. In this study, varying amounts of low molecular weight branched polyethylenimine 1.8 kDa (bPEI 1.8) were introduced on to a neutral polymer, poly(vinyl alcohol) (PVA), to bring in cationic charge on the resulting PVA-PEI (PP) nanocomposites. We rationalized that by introducing bPEI 1.8, buffering and condensation properties of the proposed nanocomposites would result in improved gene transfer capability. A series of PVA-PEI (PP) nanocomposites was synthesized using well-established epoxide chemistry and characterized by IR and NMR. Particle size of the PP/DNA complexes ranged between 120 to 135 nm, as determined by dynamic light scattering (DLS), and DNA retardation assay revealed efficient binding capability of PP nanocomposites to negatively charged nucleic acids. In vitro transfection of PP/DNA complexes in HEK293, HeLa, and CHO cells revealed that the best working formulation in the synthesized series, PP-3/DNA complex, displayed ~2-50-fold higher transfection efficiency than bPEIs (1.8 and 25 kDa) and commercial transfection reagents. More importantly, the PP/DNA complexes were stable over a period of time, along with their superior transfection efficiency in the presence of serum compared to serum-free conditions, retaining the nontoxic property of low molecular weight bPEI. The in vivo administration of PP-3/DNA complex in Balb/c mice showed maximum gene expression in their spleen. The study demonstrates the potential of PP nanocomposites as promising nonviral gene vectors for in vivo applications.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi-110007, India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: Sushil K
authors.initials: SK
authors.lastname: Tripathi
authors.forename: Esther
authors.initials: E
authors.lastname: Vazquez
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
authors.forename: Kailash C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry / Q000494:pharmacology; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053761:Nanocomposites / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry / Q000494:pharmacology; D011142:Polyvinyl Alcohol / Q000737:chemistry / Q000494:pharmacology
chemical_list: D011142:Polyvinyl Alcohol; D011094:Polyethyleneimine; D004247:DNA