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

Author(s)

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]

Affiliation

^[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

• Document type: Journal article
• Source: Nanotechnology; v. 24, n. 5 FEB 8 2013.
• Web of Science Citations: 15

Abstract

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)

• FAPESP's process: 10/09651-6 - Synthesis and Tranfection Studies of Site-Targeted Nanocarriers of Genes
• Grantee: Isadora Pfeifer Dalla Picola
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 11/18692-0 - Non-viral gene therapy: synthesis, characterization and transfection studies of site-targeted nanocarriers based on biocompatible polycations
• Grantee: Marcio José Tiera
• Support Opportunities: Regular Research Grants