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

Nature Materials volume 3, pages 249253 (2004) | Download Citation

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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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References

  1. 1.

    & Tissue engineering—current challenges and expanding opportunities. Science 295, 1009–1014 (2002).

  2. 2.

    Tissue engineering and reparative medicine. Ann. NY Acad. Sci. 961, 1–9 (2002).

  3. 3.

    , , , & Gradients of substrate-bound laminin orient axonal specification of neurons. Proc. Natl Acad. Sci. USA 99, 12542–12547 (2002).

  4. 4.

    & Patterned poly(chlorotrifluoroethylene) guides primary nerve cell adhesion and neurite outgrowth. J. Biomed. Mater. Res. 50, 465–474 (2000).

  5. 5.

    et al. Micropatterning gradients and controlling surface densities of photoactivatable biomolecules on self-assembled monolayers of oligo(ethylene glycol) alkanethiolates. Chem. Biol. 4, 731–737 (1997).

  6. 6.

    , , , & Biologically engineered protein-graft-poly(ethylene glycol) hydrogels: a cell adhesive and plasmin-degradable biosynthetic material for tissue repair. Biomacromolecules 3, 710–723 (2002).

  7. 7.

    , , & Synthesis of in situ cross-linkable macroporous biodegradable poly(propylene fumarate-co-ethylene glycol) hydrogels. Biomacromolecules 3, 374–381 (2002).

  8. 8.

    , , , & Engineering growing tissues. Proc. Natl Acad. Sci. USA 99, 12025–12030 (2002).

  9. 9.

    & Three-dimensional photopatterning of hydrogels containing living cells. Biomed. Microdevices 4, 257–266 (2002).

  10. 10.

    , & Micropatterning of biomedical polymer surfaces by novel UV polymerization techniques. J. Biomed. Mater. Res. 56, 351–360 (2001).

  11. 11.

    & Methods for the topographical patterning and patterned surface modification of hydrogels based on hydroxyethyl methacrylate. Biomacromolecules 4, 1126–1131 (2003).

  12. 12.

    & Microfluidic patterning of cellular biopolymer matrices for biomimetic 3D structures. Biomed. Microdevices 5, 235–244 (2003).

  13. 13.

    et al. Electrochemically induced deposition of a polysaccharide hydrogel onto a patterned surface. Langmuir 19, 4058–4062 (2003).

  14. 14.

    , , , & Organ printing: computer-aided jet-based 3D tissue engineering. Trends Biotechnol. 21, 157–161 (2003).

  15. 15.

    , , & Three-dimensional extracellular matrix engineering in the nervous system. J. Biomed. Mater. Res. 40, 392–400 (1998).

  16. 16.

    , , , & A methacrylated photoiniferter as a chemical basis for microlithography: Micropatterning based on photografting polymerization. Macromolecules 36, 6739–6745 (2003).

  17. 17.

    & Protein patterning. Biomaterials 19, 595–609 (1998).

  18. 18.

    , , , & Micropatterned surfaces for control of cell shape, position, and function. Biotechnol. Prog. 14, 356–363 (1998).

  19. 19.

    , & Growth cone guidance and neuron morphology on micropatterned laminin surfaces. J. Cell Sci. 105, 203–212 (1993).

  20. 20.

    , , & Guidance of neurite outgrowth by pathways of substratum-adsorbed laminin. J. Neurosci. Res. 13, 213–220 (1985).

  21. 21.

    et al. Spatial control of neuronal cell attachment and differentiation on covalently patterned laminin oligopeptide substrates. Intl J. Dev. Neurosci. 12, 725–735 (1994).

  22. 22.

    & The molecular biology of axon guidance. Science 274, 1123–1133 (1996).

  23. 23.

    , & Laminin oligopeptide derivatized agarose gels allow three-dimensional neurite extension in vitro. J. Neurosci. Res. 41, 501–509 (1995).

  24. 24.

    et al. A spinal cord surrogate with nanoscale porosity for in vitro simulations of restorative neurosurgical techniques. Nanotechnology 13, 587–591 (2002).

  25. 25.

    & Controlling cell chemistry with caged compounds. Annu. Rev. Physiol. 55, 755–784 (1993).

  26. 26.

    & Properties and uses of photoreactive caged compounds. Annu. Rev. Biophys. Biophys. Chem. 18, 239–270 (1989).

  27. 27.

    & Functional domains of cell adhesion molecules. Curr. Opin. Cell Biol. 4, 819–823 (1992).

  28. 28.

    RGD and other recognition sequences for integrins. Annu. Rev. Cell. Dev. Biol. 12, 697–715 (1996).

  29. 29.

    Lasers (University Science Books, Mill Valley, California, 1986).

  30. 30.

    in Department of Chemical Engineering and Applied Chemistry 95–96, 129–132 (University of Toronto, Toronto, Ontario, Canada, 2003).

  31. 31.

    & Physical chemistry (Wiley, New York, 2001).

  32. 32.

    & Ligand-induced changes in integrin expression regulate neuronal adhesion and neurite outgrowth. Nature 389, 852–856 (1997).

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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.

Author information

Affiliations

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

    • Ying Luo
    •  & Molly S. Shoichet
  2. Institute of Biomaterials and Biomedical Engineering, University of Toronto, 4 Taddle Creek Road, Toronto, Ontario M5S 3G9, Canada

    • Ying Luo
    •  & Molly S. Shoichet
  3. Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario M5S 3H6, Canada

    • Molly S. Shoichet

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Molly S. Shoichet.

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DOI

https://doi.org/10.1038/nmat1092

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