Improving chitosan-mediated gene transfer by the introduction of intracellular buffering moieties into the chitosan backbone

• Type: Article in International Scientific Journal
• Date: 2009

Authors

Moreira, C (Author) Other; Oliveira, H (Author) Other; Pires, LR (Author) Other; Simoes, S (Author) Other; Barbosa, MA (Author) FEUP; Pego, AP (Author) FEUP

Journal

Title: Acta Biomaterialia

Vol. 5 No. 1

Pages: 2995-3006

ISSN: 1742-7061

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 51 Citations

ISI Web of Science

Scopus - 51 Citations

INSPEC

Scientific classification

FOS: Engineering and technology > Environmental biotechnology

CORDIS: Technological sciences > Engineering > Biomaterial engineering

Other information

Authenticus ID: P-003-F3G

DOI: 10.1016/j.actbio.2009.04.021

Abstract (EN): Chitosan was functionalized with imidazole moieties (CHimi) with the aim of improving its buffering capacity and promoting the endosomal escape ability of chitosan-DNA complexes, ultimately increasing their transfection efficiency. 5.6%, 12.9% and 22.1% of the glucosamine residues of chitosan were substituted. Complexes with different molar ratios of primary amines to DNA phosphate anion (N/P) were prepared by a coacervation method. For an N/P > 3, CHimi polymers are able to complex electrostatically with DNA and condense it into positively charged nanostructures (average size 260 nm and zeta potential +16 mV at pH 5.5). In the concentration range 2.5-100 mu g ml(-1), the modified polymers had no cytotoxic effect on 293T cells. CHimi polymers with the highest degree of substitution were found to enhance beta-gal expression in 293T and HepG2 cells. Bafilomycin A1 inhibited transfection, indicating that the protonation of the imidazole groups in the endolysosome pathway favors the escape of the complexes from the endosomes, increasing the amount of transgene that can reach the cell nucleus.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 12