title: Polyethylenimine-polyacrylic acid nanocomposites: Type of bonding does influence the gene transfer efficacy and cytotoxicity.
abstract: The main aim of the current study is to compare the physicochemical properties, cytotoxicity and gene-transfer ability of electrostatically and covalently linked nanocomposites of polyethylenimine (PEI) and polyacrylic acid (PAA) on mammalian cells. Two series of nanocomposites, ionic PEI-PAA (iPP) and covalent PEI-PAA (cPP), were synthesized by varying the amounts of polyacrylic acid (PAA). Physicochemical characterization revealed that iPP nanopcomposites were of bigger sized than cPP nanocomposites with zeta potential almost comparable. Nucleic acid binding assay displayed that iPP and cPP nanocomposites, having sufficient cationic charge, efficiently interacted with plasmid DNA and completely retarded its electrophoretic mobility on agarose gel. In vitro MTT assay showed slightly higher cell viability of cPP/pDNA complexes over their ionic counterparts. Both the series of nanocomposite/pDNA complexes exhibited considerably higher transfection efficacy compared to pDNA complexes of native bPEI and the standard transfection reagent, Lipofectamine, with cPP/pDNA complexes performed much better than iPP/pDNA complexes. Flow cytometry further confirmed these findings where cPP-4/pDNA complex showed transfection in ∼ 85% HEK293 cells, while iPP-2/pDNA complex transfected ∼ 67% HEK293 cells. Lipofectamine/pDNA and bPEI/pDNA complexes could transfect just ∼ 35% and ∼ 26% HEK293 cells. All these results demonstrate the superiority of covalently linked nanocomposites (cPP) which could be used as efficient carriers for nucleic acids in future gene delivery applications.
authors.affiliation: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi-110007, India.
authors.forename: Sushil K
authors.initials: SK
authors.lastname: Tripathi
authors.affiliation: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi-110007, India.
authors.forename: Zeba
authors.initials: Z
authors.lastname: Ahmadi
authors.affiliation: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi-110007, India.
authors.forename: Kailash C
authors.initials: KC
authors.lastname: Gupta
authors.affiliation: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi-110007, India. Electronic address: pkumar@igib.in.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry*; D000818:Animals; D016466:CHO Cells; D002470:Cell Survival / Q000235:genetics; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D053761:Nanocomposites / Q000737:chemistry*; D010316:Particle Size; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D011094:Polyethyleneimine / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D014162:Transfection / Q000379:methods
chemical_list: D000180:Acrylic Resins; D049452:Green Fluorescent Proteins; C006903:carbopol 940; D011094:Polyethyleneimine; D004247:DNA
keywords: Cytotoxicity; Non-viral vector; Polyacrylic acid; Polyethylenimine; Transfection efficiency