Pharmaceutical Research

, Volume 15, Issue 9, pp 1332–1339

Chitosan-Based Vector/DNA Complexes for Gene Delivery: Biophysical Characteristics and Transfection Ability

  • Patrick Erbacher
  • Shaomin Zou
  • Thierry Bettinger
  • Anne-Marie Steffan
  • Jean-Serge Remy


Purpose. Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery. With the aim of developing this system, various biophysical characteristics of chitosan-condensed DNA complexes were measured, and transfections were performed.

Methods. Transmission electronic microscopy (TEM) visualizations, sedimentation experiments, dynamic light scattering (DLS), and zeta potential measurements were realized. Transfections were made by using the luciferase reporter gene.

Results. In defined conditions, plasmid DNA formulated with chitosan produced homogenous populations of complexes which were stable and had a diameter of approximately 50−100 nm. Discrete particles of nicely condensed DNA had a donut, rod, or even pretzel shape. Chitosan/DNA complexes efficiently transfected HeLa cells, independently of the presence of 10% serum, and did not require an added endosomolytic agent. In addition, gene expression gradually increased over time, from 24 to 96 hours, whereas in the same conditions the efficacy of polyethylenimine-mediated transfection dropped by two orders of magnitude. At 96 hours, chitosan was found to be 10 times more efficient than PEI. However, chitosan-mediated transfection depended on the cell type. This dependency is discussed here.

Conclusions. Chitosan presents some characteristics favorable for gene delivery, such as the ability to condense DNA and form small discrete particles in defined conditions.

gene therapy gene transfer cationic polymer chitosan polyethylenimine 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    F. D. Ledley. Pharmaceutical approach to somatic gene therapy. Pharm. Res. 13:1595–1614 (1996).PubMedGoogle Scholar
  2. 2.
    G. Y. Wu and C. H. Wu. Receptor-mediated in vitro gene transformation by a soluble DNA carrier system. J. Biol. Chem. 262:4429–4432 (1987).PubMedGoogle Scholar
  3. 3.
    J. Haensler and F. C. Szoka Jr. Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjugate Chem. 4:372–379 (1993).Google Scholar
  4. 4.
    O. Boussif, F. Lezoualc'h, M. A. Zanta, M. D. Mergny, D. Scherman, B. Demeneix, and J. P. Behr. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc. Natl. Acad. Sci. USA 92:7297–7301 (1995).PubMedGoogle Scholar
  5. 5.
    M. X. Tang and F. C. Szoka. The influence of polymer structure on the interactions of cationic polymers with DNA and morphology of the resulting complexes. Gene Ther. 4:823–832 (1997).PubMedGoogle Scholar
  6. 6.
    J. P. Behr, B. Demeneix, J. P. Loeffler, and J. Perez-Mutul. Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA. Proc. Natl. Acad. Sci. USA 89:6982–6986 (1989).Google Scholar
  7. 7.
    K. A. Mislick and J. D. Baldeschwieler. Evidence for the role of proteoglycans in cation-mediated gene transfer. Proc. Natl. Acad. Sci. USA 93:12349–12354 (1996).PubMedGoogle Scholar
  8. 8.
    F. Labat-Moleur, A. M. Steffan, C. Brisson, H. Perron, O. Feugeas, P. Furstenberger, F. Oberling, E. Brambilla, and J. P. Behr. An electron microscopy study into the mechanism of gene transfer with lipopolyamines. Gene Ther. 3:1010–1017 (1996).PubMedGoogle Scholar
  9. 9.
    K. Roy, H. Q. Mao, and K. W. Leong. DNA-chitosan nanospheres: transfection efficiency and cellular uptake. Proceed. Int'l. Symp. Control. Rel. Bioact. Mater. 24:673–674 (1997).Google Scholar
  10. 10.
    J. L. Murata, Y. Ohya, and T. Ouchi. Possibility of application of quaternary chitosan having pendant galactose residues as gene delivery tool. Carbohydrate Polymers 29:69–74 (1996).Google Scholar
  11. 11.
    M. Monsigny, C. Petit, and A. C. Roche. Colorimetric determination of neutral sugars by a recorcinol sulfuric acid micromethod. Anal. Biochem. 175:525–530 (1988).PubMedGoogle Scholar
  12. 12.
    M. A. Zanta, O. Boussif, A. Adib, and J. P. Behr. In vitro gene delivery to hepatocytes with galactosylated polyethylenimine. Bioconjugate Chem 8:839–844 (1997).Google Scholar
  13. 13.
    C. K. Goldman, L. Soroceanu, N. Smith, Y. Gillepsie, W. Shaw, S. Burgess, G. Bilbao, and D. T. Curiel. In vitro and in vivo gene delivery mediated by a synthetic polycationic amino polymer. Nature Biotech. 15:462–466 (1997).Google Scholar
  14. 14.
    M. Cotten, F. Längle-Rouault, H. Kirlapos, E. Wagner, K. Mechtler, M. Zenke, H. Beug, and M. L. Birnstiel. Transferrin-polycation-mediated introduction of DNA into human leukemic cells: stimulation by agents that affect the survival of transfected DNA or modulate tansferrin receptor levels. Proc. Natl. Acad. Sci. USA 87:4033–4037 (1990).PubMedGoogle Scholar
  15. 15.
    P. Midoux, C. Mendès, A. Legrand, J. Raimond, R. Mayer, M. Monsigny, and A. C. Roche. Specific gene transfer mediated by lactosylated poly-L-lysine into hepatoma cells. Nucleic Acids Res. 21:871–878 (1993).PubMedGoogle Scholar
  16. 16.
    A. S. Chuck, M. F. Clarke, and B. O. Palsson. Retroviral infection is limited by brownian motion. Human Gene Ther. 7:1527–1534 (1996).Google Scholar
  17. 17.
    D. T. Curiel, S. Agarwal, E. Wagner, and M. Cotten. Adenovirus enhancement of transferrin-polylysine-mediated gene delivery. Proc. Natl. Acad. Sci. USA 88:8850–8854 (1991).PubMedGoogle Scholar
  18. 18.
    C. Plank, K. Zatloukal, M. Cotten, K. Mechtler, and E. Wagner. Gene transfer into hepatocytes using asialoglycoprotein receptor mediated endocytosis of DNA complexed with an artificial tetra-antennary galactose ligand. Bioconjugate Chem. 3:533–539. (1992).Google Scholar
  19. 19.
    C. Planck, B. Oberhauser, K. Mechtler, C. Koch, and E. Wagner. The influence of endosome-disruptive peptides on gene transfer using synthetic virus-like gene transfer system. J. Biol. Chem. 269:12918–12924 (1994).PubMedGoogle Scholar
  20. 20.
    P. Erbacher, A. C. Roche, M. Monsigny, and P. Midoux. Putative role of chloroquine in gene transfer into a hepatoma cell line by DNA/lactosylated polylysine complexes. Exp. Cell Res. 225:186–194 (1996).PubMedGoogle Scholar
  21. 21.
    J. P. Behr. The proton sponge: a trick to enter cells the viruses did not think of. Chimia 51:27–30 (1997).Google Scholar

Copyright information

© Plenum Publishing Corporation 1998

Authors and Affiliations

  • Patrick Erbacher
    • 1
  • Shaomin Zou
    • 1
  • Thierry Bettinger
    • 1
  • Anne-Marie Steffan
    • 2
  • Jean-Serge Remy
    • 1
  1. 1.Laboratoire de Chimie GénétiqueUniversité Louis-Pasteur de StrasbourgIllkirchFrance
  2. 2.Institut de VirologieStrasbourgFrance

Personalised recommendations