Synthesis and evaluation of diethylethylamine-chitosan for gene delivery: composition effects on the in vitro transfection efficiency

Autor(es)

Pansani Oliveira, Franciele de Paula ^[1] ; Dalla Picola, Isadora Pfeifer ^{[1, 2, 3]} ; Shi, Qin ^[2] ; Goncalves Barbosa, Hellen Franciane ^[1] ; de Oliveira Tiera, Vera Aparecida ^[1] ; Fernandes, Julio Cesar ^[2] ; Tiera, Marcio Jose ^[1]

Afiliação do(s) autor(es)

^[1] Sao Paulo State Univ UNESP, IBILCE, Dept Chem & Environm Sci, Sao Paulo - Brazil

^[2] Univ Montreal, Hop Sacre Coeur Montreal, Orthoped Res Lab, Montreal, PQ H3C 3J7 - Canada

^[3] Sao Paulo State Univ UNESP, IBILCE, Dept Phys, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Nanotechnology; v. 24, n. 5 FEB 8 2013.
• Citações Web of Science: 15

Resumo

Chitosan has been indicated as a safe and promising polycation vector for gene delivery. However its low transfection efficiency has been a challenging obstacle for its application. To address this limitation, we synthesized chitosan derivatives which had increasing amounts of diethylethylamine groups (DEAE) attached to the chitosan main chain. The plasmid DNA VR1412 (pDNA), encoding the beta-galactosidase (beta-gal) reporter gene was used to prepare nanoparticles with the chitosan derivatives, and the transfection studies were performed with HeLa cells. By means of dynamic light scattering and zeta potential measurements, it was shown that diethylethylamine-chitosan derivatives (DEAE(x)-CH) were able to condense DNA into small particles having a surface charge depending on the polymer/DNA ratio (N/P ratio). Nanoparticles prepared with derivatives containing 15 and 25% of DEAE groups (DEAE(15)-CH and DEAE(25)-CH) exhibited transfection efficiencies ten times higher than that observed with deacetylated chitosan (CH). For derivatives with higher degrees of substitution (DS), transfection efficiency decreased. The most effective carriers showed low cytotoxicity and good transfection activities at low charge ratios (N/P). Vectors with low DS were easily degraded in the presence of lysozyme at physiological conditions in vitro and the nontoxicity displayed by these vectors opens up new opportunities in the design of DEAE-chitosan-based nanoparticles for gene delivery. (AU)

• Processo FAPESP: 10/09651-6 - Síntese e Estudos de Transfecção de nanocarreadores sítio-dirigidos de genes
• Beneficiário: Isadora Pfeifer Dalla Picola
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 11/18692-0 - Terapia gênica não-viral: síntese, caracterização e estudos de transfecção de nanocarreadores sítio-dirigidos de genes baseados em policátions biocompatíveis
• Beneficiário: Marcio José Tiera
• Modalidade de apoio: Auxílio à Pesquisa - Regular