title: Effect of homobifunctional crosslinkers on nucleic acids delivery ability of PEI nanoparticles.
abstract: Polyethylenimine (PEI), a widely used cationic polymeric vector with high transfection efficiency, was converted into nanoparticles by introducing ionic and covalent crosslinkers with varying proportion of 1,6-hexanebisphosphate (HP), adipic acid (AA) and 1,4-butane dialdehyde (BA) to obtain a small library of HP-PEI (HPP), AA-PEI (AAP) and BA-PEI (BAP) nanoparticles, respectively. Particles were characterized by spectroscopic technique as well as physicochemical parameters such as size, morphology, surface charge, effect of crosslinking on buffering capacity and DNA binding ability. The entire series of nanoparticles were compared for their cytoxicity and ability to deliver genes in various cell lines. Among various nanoparticles, AAP-3 nanoparticle/DNA complex exhibited higher transfection efficiency (1.5-7.8 folds) than the native PEI (25kDa) and commercially available transfection reagents, such as GenePorter, GenePorter2, Fugene and Superfect, with cell viability >85%. The highest cell viability was observed with BAP nanoparticles (>95%). Importantly, the transfection activity of nanoparticle/DNA complexes was preserved in the presence of serum. Transfection with GFP-siRNA inhibited expression of transfected GFP gene by approximately 81-92%. All nanoparticle types (HPP, AAP and BAP) required a comparable time for entry into cells and subsequent intracellular passage from the cytoplasm to the nucleus. Intravenous delivery of (99)Tc labeled BAP-2/DNA complex to female Balb/c mice revealed the presence of the complex in most of the organs with the highest retention in liver. In conclusion, HPP, AAP and BAP nanoparticles are safe for efficient gene delivery.
authors.affiliation: Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi, India.
authors.forename: Archana
authors.initials: A
authors.lastname: Swami
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: Sushil Kumar
authors.initials: SK
authors.lastname: Tripathi
authors.forename: Naresh
authors.initials: N
authors.lastname: Singh
authors.forename: Neeraj
authors.initials: N
authors.lastname: Katiyar
authors.forename: Anil K
authors.initials: AK
authors.lastname: Mishra
authors.forename: K C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000272:Adipates / Q000737:chemistry; D000447:Aldehydes / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage* / Q000493:pharmacokinetics; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles; D009943:Organophosphorus Compounds / Q000737:chemistry; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000493:pharmacokinetics; D013499:Surface Properties; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods
chemical_list: D000272:Adipates; D000447:Aldehydes; D003432:Cross-Linking Reagents; D009943:Organophosphorus Compounds; D034741:RNA, Small Interfering; C015515:succindialdehyde; C018313:1,6-hexanediol bisphosphate; C029900:adipic acid; D011094:Polyethyleneimine; D004247:DNA