Imidazole-grafted chitosan-mediated gene delivery: in vitro study on transfection, intracellular trafficking and degradation.
OBJECTIVE
To study the mechanism of transfection mediated by imidazole-grafted chitosan (CHimi) nanoparticles, to propose new strategies to control and improve the expression of a delivered gene in the context of regenerative medicine.


METHODS
Biochemical and microscopy methods were used to establish transfection efficiency and nanoparticle intracellular trafficking. The role of CHimi and degree of N-acetylation (DA) on transfection was explored.


RESULTS
CHimi was found to promote the expression of a delivered gene during a minimum 7-day period. Additionally, the production of a protein of interest could be upheld by consecutive transfections, without compromising cell viability. Transfection was found to be a time-dependent process, requiring CHimi-DNA complex disassembling. The DA was found to have an impact on transfection kinetics in line with the observation that the rate of lysozyme-mediated nanoparticle degradation increases with the polymer DA.


CONCLUSIONS
The adjustment of the CH degradation rate can be used as a tool for tuning the expression of a gene delivered by CH-based nanoparticle systems.
Instituto de Engenharia Biomédica, Divisão de Biomateriais, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal.
Liliana R
LR
Pires
Hugo
H
Oliveira
Cristina C
CC
Barrias
Paula
P
Sampaio
António J
AJ
Pereira
Hélder
H
Maiato
Sérgio
S
Simões
Ana P
AP
Pêgo
D002460:Cell Line; D048271:Chitosan / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006801:Humans; D007093:Imidazoles / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D014162:Transfection / Q000379:methods*
D007093:Imidazoles; C029899:imidazole; D048271:Chitosan