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

• Tipo: Artigo em Revista Científica Internacional
• Data: 2009

Autores

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

Revista

Título: Acta Biomaterialia

Vol. 5 Nº 1

Páginas: 2995-3006

ISSN: 1742-7061

Editora: Elsevier

Indexação

ISI Web of Knowledge - 51 Citações

ISI Web of Science

Scopus - 51 Citações

INSPEC

Classificação Científica

FOS: Ciências da engenharia e tecnologias > Biotecnologia ambiental

CORDIS: Ciências Tecnológicas > Engenharia > Engenharia de biomateriais

Outras Informações

ID Authenticus: 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 12