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Non-viral gene delivery regulated by stiffness of cell adhesion substrates

Nature Materials volume 4, pages 460–464 (2005)Cite this article

Abstract

Non-viral gene vectors are commonly used for gene therapy1,2,3 owing to safety concerns with viral vectors4. However, non-viral vectors are plagued by low levels of gene transfection and cellular expression1,2. Current efforts to improve the efficiency of non-viral gene delivery are focused on manipulations of the delivery vector5,6,7,8,9,10,11,12, whereas the influence of the cellular environment in DNA uptake is often ignored. The mechanical properties (for example, rigidity) of the substrate to which a cell adheres have been found to mediate many aspects of cell function including proliferation, migration and differentiation13,14,15,16,17, and this suggests that the mechanics of the adhesion substrate may regulate a cell's ability to uptake exogeneous signalling molecules. In this report, we present a critical role for the rigidity of the cell adhesion substrate on the level of gene transfer and expression. The mechanism relates to material control over cell proliferation, and was investigated using a fluorescent resonance energy transfer (FRET) technique18,19,20,21. This study provides a new material-based control point for non-viral gene therapy.

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Figure 1: The interaction between pDNA fluorescently labelled with Alexa 488 and PEI labelled with rhodamine.
Figure 2: Uptake of the pDNA–PEI condensates into the cells.
Figure 3: The levels of gene transfer and expression were dependent on the stiffness of cell-adhesive hydrogels.
Figure 4: Cellular proliferation and apoptosis were regulated by the elastic moduli (E) of the gels.
Figure 5: Gene expression was examined over a broad range of mechanical properties using several biomaterials conventionally used for in vitro cell culture.

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Acknowledgements

We thank Michael Solomon at the University of Michigan for use of the laser scanning confocal microscope. We also thank Yoshiaki Hirano at Osaka University for helpful discussions. Financial support was provided by the National Institute of Health (R30 DE 13033).

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Authors and Affiliations

  1. Division of Engineering and Applied Sciences, Harvard University, Cambridge, 02138, Massachusetts, USA

    Hyun Joon Kong, Takuya Matsumoto, Kent Leach & David J. Mooney

  2. Chemical Engineering, University of Michigan, Ann Arbor, 48109, Michigan, USA

    Jodi Liu & Kathryn Riddle

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Correspondence to David J. Mooney.

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Kong, H., Liu, J., Riddle, K. et al. Non-viral gene delivery regulated by stiffness of cell adhesion substrates. Nature Mater 4, 460–464 (2005). https://doi.org/10.1038/nmat1392

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