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A photolabile hydrogel for guided three-dimensional cell growth and migration

Nature Materials volume 3, pages 249–253 (2004)Cite this article

Abstract

Tissue engineering aims to replace, repair or regenerate tissue/organ function, by delivering signalling molecules and cells on a three-dimensional (3D) biomaterials scaffold that supports cell infiltration and tissue organization1,2. To control cell behaviour and ultimately induce structural and functional tissue formation on surfaces, planar substrates have been patterned with adhesion signals that mimic the spatial cues to guide cell attachment and function3,4,5. The objective of this study is to create biochemical channels in 3D hydrogel matrices for guided axonal growth. An agarose hydrogel modified with a cysteine compound containing a sulphydryl protecting group provides a photolabile substrate that can be patterned with biochemical cues. In this transparent hydrogel we immobilized the adhesive fibronectin peptide fragment, glycine–arginine–glycine–aspartic acid–serine (GRGDS), in selected volumes of the matrix using a focused laser. We verified in vitro the guidance effects of GRGDS oligopeptide-modified channels on the 3D cell migration and neurite outgrowth. This method for immobilizing biomolecules in 3D matrices can generally be applied to any optically clear hydrogel, offering a solution to construct scaffolds with programmed spatial features for tissue engineering applications.

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Figure 1: The general strategy used to create adhesive biochemical channels in agarose hydrogel matrices relies on modifying agarose with photolabile groups, focused laser light sources and biomolecule coupling.
Figure 2: Biochemical channels synthesized in agarose hydrogels were characterized with a fluorescein-tagged GRGDS peptide.
Figure 3: The transmittance of laser light is affected by both the thicknesses of the S-NBC-modified agarose hydrogel matrices and the concentration of S-NBC of 0.4 mM and 0.8 mM (mean ± standard deviation, n = 5).
Figure 4: Primary rat dorsal root ganglia cells were plated on 3D patterned GRGDS oligopeptide-modified, 0.5 wt% agarose gels.

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Acknowledgements

We are grateful to the Natural Sciences and Engineering Research Council of Canada, Ontario Graduate Scholarship and Connaught for funding and thank Ying-Fang Chen, Patricia Musoke-Zawedde and David Martens for their assistance.

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Authors and Affiliations

  1. Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, M5S 3E5, Ontario, Canada

    Ying Luo & Molly S. Shoichet

  2. Institute of Biomaterials and Biomedical Engineering, University of Toronto, 4 Taddle Creek Road, Toronto, M5S 3G9, Ontario, Canada

    Ying Luo & Molly S. Shoichet

  3. Department of Chemistry, University of Toronto, Toronto, M5S 3H6, 80 St. George Street, Ontario, Canada

    Molly S. Shoichet

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  1. Ying Luo
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Correspondence to Molly S. Shoichet.

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Luo, Y., Shoichet, M. A photolabile hydrogel for guided three-dimensional cell growth and migration. Nature Mater 3, 249–253 (2004). https://doi.org/10.1038/nmat1092

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