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Chitosan as a nonviral gene delivery system. Structure–property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo

Gene Therapy volume 8, pages 1108–1121 (2001)Cite this article

Abstract

Chitosan is a natural cationic linear polymer that has recently emerged as an alternative nonviral gene delivery system. We have established the relationships between the structure and the properties of chitosan-pDNA polyplexes in vitro. Further, we have compared polyplexes of ultrapure chitosan (UPC) of preferred molecular structure with those of optimised polyethylenimine (PEI) polyplexes in vitro and after intratracheal administration to mice in vivo. Chitosans in which over two out of three monomer units carried a primary amino group formed stable colloidal polyplexes with pDNA. Optimized UPC and PEI polyplexes protected the pDNA from serum degradation to approximately the same degree, and they gave a comparable maximal transgene expression in 293 cells. In contrast to PEI, UPC was non toxic at escalating doses. After intratracheal administration, both polyplexes distributed to the mid-airways, where transgene expression was observed in virtually every epithelial cell, using a sensitive pLacZ reporter containing a translational enhancer element. However, the kinetics of gene expression differed – PEI polyplexes induced a more rapid onset of gene expression than UPC. This was attributed to a more rapid endosomal escape of the PEI polyplexes. Although this resulted in a more efficient gene expression with PEI polyplexes, UPC had an efficiency comparable to that of commonly used cationic lipids. In conclusion, this study provides insights into the use of chitosan as a gene delivery system. It emphasises that chitosan is a nontoxic alternative to other cationic polymers and it forms a platform for further studies of chitosan-based gene delivery systems.

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Acknowledgements

We would like to thank Valentis, Inc., Burlingame, CA, USA for the gift of pCMV-CAT, pCMV-Luc and fluorescently labelled pDNA, Dr Göran Ocklind, Dr Lucia Lazorova, Tapio Nikkilä, Anna Johansson and Björn Jansson for skilful technical assistance and Professor Godfried Roomans for help with analysis of lung sections. This work was supported by a grant from the Wallenberg Foundation.

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Authors and Affiliations

  1. Department of Pharmacy, Division of Pharmaceutics, Uppsala University, Uppsala, Sweden

    M Köping-Höggård, I Tubulekas, H Guan, M Nilsson & P Artursson

  2. Department of Physical Chemistry, Uppsala University, Uppsala, Sweden

    K Edwards

  3. Department of Biotechnology, Norwegian Biopolymer Laboratory (NOBIPOL), The Norwegian University of Science and Technology, Trondheim, Norway

    KM Vårum

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Köping-Höggård, M., Tubulekas, I., Guan, H. et al. Chitosan as a nonviral gene delivery system. Structure–property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo. Gene Ther 8, 1108–1121 (2001). https://doi.org/10.1038/sj.gt.3301492

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