title: Crosslinked nanocarriers based upon poly(ethylene imine) for systemic plasmid delivery: in vitro characterization and in vivo studies in mice.
abstract: Crosslinked poly(ethylene imine) (PEI) polyplexes for intracellular DNA release were generated using a low molecular weight crosslinking reagent, Dithiobis(succinimidyl propionate) (DSP). Disulfide bonds of the crosslinked polyplexes were susceptible to intracellular redox conditions and DNA release was observed using an ethidium bromide exclusion assay and dynamic light scattering. Transfection experiments were performed to elucidate the effect of extra- and intracellular redox conditions. Pharmacokinetics and organ accumulation of uncrosslinked and crosslinked polyplexes were compared and gene expression patterns were measured in mice 24 h after intravenous injection. Crosslinked PEI and plasmid DNA formed stable polyplexes in a size range of 100-300 nm, with zeta potentials between +16.4 and +26.1 mV. DNA release occurred after cleavage of the disulfide bonds. Cell culture experiments under reducing conditions as well as with glutathione loaded cells confirmed the proposed intracellular activation. A significant influence of the intracellular glutathione status on the transfection efficiency was observed. Pharmacokinetic profiles of crosslinked PEI/DNA polyplexes in mice after intravenous administration showed higher blood levels for crosslinked polyplexes. These polyplexes accumulated mainly in the liver and the lungs. In vivo transfection data revealed significantly reduced (unwanted) lung transfection while liver transfection predominated. These studies suggest that crosslinked polyplexes are more stable in circulation and retain their transfection efficiency after intravenous administration.
authors.affiliation: Department of Pharmaceutics and Biopharmacy, Philipps Universität, Ketzerbach 63, Marburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Neu
authors.forename: Oliver
authors.initials: O
authors.lastname: Germershaus
authors.forename: Shirui
authors.initials: S
authors.lastname: Mao
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Voigt
authors.forename: Martin
authors.initials: M
authors.lastname: Behe
authors.forename: Thomas
authors.initials: T
authors.lastname: Kissel
mesh_terms: D000818:Animals; D003432:Cross-Linking Reagents / Q000008:administration & dosage* / Q000737:chemistry; D018014:Gene Transfer Techniques; D051379:Mice; D041681:NIH 3T3 Cells; D053769:Nanocapsules / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D011094:Polyethyleneimine / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D053769:Nanocapsules; D011094:Polyethyleneimine