title: The influence of physicochemical parameters on the efficacy of non-viral DNA transfection complexes: a comparative study.
abstract: Various polycationic vehicles have been developed to facilitate the transfer of foreign DNA into mammalian cells. Structure-activity studies suggested that biophysical properties, such as size, charge, and morphology of the resulting DNA complexes determine transfection efficiency within one class of vector. To investigate the general validity of these criteria, we studied the efficacy of a variety of DNA delivery vehicles including liposomes (DOTAP, SAINT2) with and without helper lipid (DOPE), the polymer polyethyleneimine (PEI), and cationic nanoparticles (Si26H, PLGA/chitosan) in a comparative manner. Sizes of the DNA complexes varied between 100 and 500 nm for PEI polyplexes and DOTAP/DOPE lipoplexes, respectively. The zeta potential was positive for PEI, Si26H, and DOTAP based complexes, while it was neutral for SAINT2-DNA complexes and negative for PLGA/chitosan-DNA complexes. The latter finding was elucidated by AFM, showing a layer of DNA adsorbed onto the nanoparticles. Transfection activity was negligible for PLGA/chitosan nanospheres, moderate for Si26H nanospheres and high for all other complexes, PEI being the most active carrier. The liposomal preparations were of low (DOTAP) or moderate (SAINT2) stability in serum, resulting in a pronounced reduction of gene expression, which was partially restored by the addition of chloroquine. In conclusion, transfection efficiency (i) seems to require a positive or neutral zeta potential, (ii) is depending on size, e.g., is higher for smaller particles, and (iii) requires a vector that is stable in serum.
authors.affiliation: Department of Pharmaceutical Technology, Philipps University Marburg, 35032 Marburg, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Kneuer
authors.forename: Carsten
authors.initials: C
authors.lastname: Ehrhardt
authors.forename: Heike
authors.initials: H
authors.lastname: Bakowsky
authors.forename: M N V Ravi
authors.initials: MN
authors.lastname: Kumar
authors.forename: Volker
authors.initials: V
authors.lastname: Oberle
authors.forename: Claus M
authors.initials: CM
authors.lastname: Lehr
authors.forename: Dick
authors.initials: D
authors.lastname: Hoekstra
authors.forename: Udo
authors.initials: U
authors.lastname: Bakowsky
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004247:DNA / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics*; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry* / Q000493:pharmacokinetics*; D005822:Genetic Vectors; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry* / Q000493:pharmacokinetics*; D008422:Materials Testing; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000235:genetics
chemical_list: D004337:Drug Carriers; D008081:Liposomes; D004247:DNA