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Biotechnology Letters

, Volume 37, Issue 3, pp 557–565 | Cite as

Improved stability and efficacy of chitosan/pDNA complexes for gene delivery

  • Noemi Cifani
  • Laura Chronopoulou
  • Barbara Pompili
  • Antonio Di Martino
  • Federico Bordi
  • Simona Sennato
  • Enea Gino Di Domenico
  • Cleofe PalocciEmail author
  • Fiorentina Ascenzioni
Original Research Paper

Abstract

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.

Keywords

Chitosan Chitosan/pDNA nanocomplexes DNA transfection Gene delivery HEK293 cells Transfection efficiency Therapeutic genes 

Notes

Acknowledgements

Istituto Pasteur Fondazione Cenci Bolognetti is acknowledged for financial support to FA and a Fellowship to EGD. LC, CP, SS and FB acknowledge financial support from Istituto Italiano di Tecnologia (IIT) Center of Life NanoScience-Sapienza. Conceived and designed the experiments: FA, CP, FB. Performed the experiments: NC, LC, BP, AM, SS. Analyzed the data: NC, LC, SS, EGD, FA, CP, FB. Wrote the paper: FA, CP.

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Noemi Cifani
    • 1
  • Laura Chronopoulou
    • 2
  • Barbara Pompili
    • 1
  • Antonio Di Martino
    • 2
    • 3
  • Federico Bordi
    • 4
    • 5
  • Simona Sennato
    • 4
    • 5
  • Enea Gino Di Domenico
    • 1
  • Cleofe Palocci
    • 2
    Email author
  • Fiorentina Ascenzioni
    • 1
  1. 1.“C. Darwin” Department of Biology and Biotechnology, Institute Pasteur-Fondazione Cenci Bolognettic/o “Sapienza” University of RomeRomeItaly
  2. 2.Department of Chemistry“La Sapienza” University of RomeRomeItaly
  3. 3.Faculty of Technology, University Institute and Polymer CentreTomas Bata University in ZlinZlinCzech Republic
  4. 4.Department of Physics“Sapienza” University of RomeRomeItaly
  5. 5.CNR-IPCF UOS Roma, c/o Department of Physics, “Sapienza” University of RomeRomeItaly

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