Improved stability and efficacy of chitosan/pDNA complexes for gene delivery.
Among polymeric polycations, chitosan has emerged as a powerful carrier for gene delivery. Only a few studies have focused on the stability of the chitosan/DNA complex under storage, although this is imperative for nanomedicinal applications. Here, we synthesized polyelectrolyte complexes at a charge ratio of 10 using 50 kDa chitosan and plasmid (p)DNA that encodes a GFP reporter. These preparations were stable up to 3 months at 4 °C and showed reproducible transfection efficiencies in vitro in HEK293 cells. In addition, we developed a methodology that increases the in vitro transfection efficiency of chitosan/pDNA complexes by 150% with respect to standard procedures. Notably, intracellular pDNA release and transfected cells peaked 5 days following transection of mitotically active cells. These new developments in formulation technology enhance the potential for polymeric nanoparticle-mediated gene therapy.
"C. Darwin" Department of Biology and Biotechnology, Institute Pasteur-Fondazione Cenci Bolognetti, c/o "Sapienza" University of Rome, P.le Aldo Moro 5, 00185, Rome, Italy.
Noemi
N
Cifani
Laura
L
Chronopoulou
Barbara
B
Pompili
Antonio
A
Di Martino
Federico
F
Bordi
Simona
S
Sennato
Enea Gino
EG
Di Domenico
Cleofe
C
Palocci
Fiorentina
F
Ascenzioni
D002460:Cell Line; D048271:Chitosan / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004355:Drug Stability; D004847:Epithelial Cells / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D010957:Plasmids; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic
D004247:DNA; D048271:Chitosan