title: Chitosan as a non-viral co-transfection system in a cystic fibrosis cell line.
abstract: Successful gene therapy requires the development of suitable vehicles for the selective and efficient delivery of genes to specific target cells at the expense of minimal toxicity. In this work, we investigated a non-viral gene delivery system based on chitosan (CS) to specifically address cystic fibrosis (CF). Thus, electrostatic self-assembled CS-pEGFP and CS-pEGFP-siRNA complexes were prepared from high-pure fully characterized CS (Mw ∼ 20 kDa and degree of acetylation ∼ 30%). The average diameter of positively-charged complexes (i.e. ζ ∼+25 mV) was ∼ 200 nm. The complexes were found relatively stable over 14h in Opti-MEM. Cell viability study did not show any significant cytotoxic effect of the CS-based complexes in a human bronchial cystic fibrosis cell line (CFBE41o-). We evaluated the transfection efficiency of this cell line with both CS-pEGFP and co-transfected with CS-pEGFP-siRNA complexes at (N/P) charge ratio of 12. We reported an increase in the fluorescence intensity of CS-pEGFP and a reduction in the cells co-transfected with CS-pEGFP-siRNA. This study shows proof-of-principle that co-transfection with chitosan might be an effective delivery system in a human CF cell line. It also offers a potential alternative to further develop therapeutic strategies for inherited disease treatments, such as CF.
authors.affiliation: Institute of Animal Physiology, University of Münster, Schlossplatz 8, D-48143 Münster, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Fernández Fernández
authors.affiliation: Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149 Münster, Germany.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Santos-Carballal
authors.affiliation: Institute of Animal Physiology, University of Münster, Schlossplatz 8, D-48143 Münster, Germany. Electronic address: wmw@uni-muenster.de.
authors.forename: Wolf-Michael
authors.initials: WM
authors.lastname: Weber
authors.affiliation: Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149 Münster, Germany. Electronic address: goycoole@uni-muenster.de.
authors.forename: Francisco M
authors.initials: FM
authors.lastname: Goycoolea
mesh_terms: D002460:Cell Line; D048271:Chitosan / Q000008:administration & dosage* / Q000737:chemistry; D003550:Cystic Fibrosis; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D010957:Plasmids; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D048271:Chitosan
keywords: Chitosan; Cystic fibrosis; Gene delivery; Nanoparticles; Transfection