Abstract
To provide insight into how cells receive information from their external surroundings, synthetic hydrogels have emerged as systems for assaying cell function in well-defined microenvironments where single cues can be introduced and subsequent effects individually elucidated. However, as answers to more complex biological questions continue to be sought, advanced material systems are needed that allow dynamic alteration of the three-dimensional cellular environment with orthogonal reactions that enable multiple levels of control of biochemical and biomechanical signals. Here, we seek to synthesize one such three-dimensional culture system using cytocompatible and wavelength-specific photochemical reactions to create hydrogels that allow orthogonal and dynamic control of material properties through independent spatiotemporally regulated photocleavage of crosslinks and photoconjugation of pendant functionalities. The results demonstrate the versatile nature of the chemistry to create programmable niches to study and direct cell function by modifying the local hydrogel environment.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Kolb, H. C., Finn, M. G. & Sharpless, K. B. Click chemistry: diverse chemical function from a few good reactions. Angew. Chem. Int. Ed. 40, 2004–2021 (2001).
Kolb, H. C. & Sharpless, K. B. The growing impact of click chemistry on drug discovery. Drug Discov. Today 8, 1128–1137 (2003).
Moses, J. E. & Moorhouse, A. D. The growing applications of click chemistry. Chem. Soc. Rev. 36, 1249–1262 (2007).
Hawker, C. J. & Wooley, K. L. The convergence of synthetic organic and polymer chemistries. Science 309, 1200–1205 (2005).
Barner-Kowollik, C. et al. ‘Clicking’ polymers or just efficient linking: what is the difference? Angew. Chem. Int. Ed. 50, 60–62 (2011).
Sletten, E. M. & Bertozzi, C. R. Bioorthogonal chemistry: fishing for selectivity in a sea of functionality. Angew. Chem. Int. Ed. 48, 6974–6998 (2009).
Bowman, C. N. & Kloxin, C. J. Toward an enhanced understanding and implementation of photopolymerization reactions. AIChE J. 54, 2775–2795 (2008).
DeForest, C. A., Polizzotti, B. D. & Anseth, K. S. Sequential click reactions for synthesizing and patterning three-dimensional cell microenvironments. Nature Mater. 8, 659–664 (2009).
Luo, Y. & Shoichet, M. S. A photolabile hydrogel for guided three-dimensional cell growth and migration. Nature Mater. 3, 249–253 (2004).
Aizawa, Y., Wylie, R. & Shoichet, M. Endothelial cell guidance in 3D patterned scaffolds. Adv. Mater. 22, 4831–4835 (2010).
Lee, S. H., Moon, J. J. & West, J. L. Three-dimensional micropatterning of bioactive hydrogels via two-photon laser scanning photolithography for guided 3D cell migration. Biomaterials 29, 2962–2968 (2008).
Hoffmann, J. C. & West, J. L. Three-dimensional photolithographic patterning of multiple bioactive ligands in poly(ethylene glycol) hydrogels. Soft Matter 6, 5056–5063 (2010).
Seidlits, S. K., Schmidt, C. E. & Shear, J. B. High-resolution patterning of hydrogels in three dimensions using direct-write photofabrication for cell guidance. Adv. Funct. Mater. 19, 3543–3551 (2009).
Kloxin, A. M., Kasko, A. M., Salinas, C. N. & Anseth, K. S. Photodegradable hydrogels for dynamic tuning of physical and chemical properties. Science 324, 59–63 (2009).
Khetan, S., Katz, J. S. & Burdick, J. A. Sequential crosslinking to control cellular spreading in 3-dimensional hydrogels. Soft Matter 5, 1601–1606 (2009).
Khetan, S. & Burdick, J. A. Patterning network structure to spatially control cellular remodeling and stem cell fate within 3-dimensional hydrogels. Biomaterials 31, 8228–8234 (2010).
Sarig-Nadir, O., Livnat, N., Zajdman, R., Shoham, S. & Seliktar, D. Laser photoablation of guidance microchannels into hydrogels directs cell growth in three dimensions. Biophys. J. 96, 4743–4752 (2009).
Ilina, O., Bakker, G. J., Vasaturo, A., Hofmann, R. M. & Friedl, P. Two-photon laser-generated microtracks in 3D collagen lattices: principles of mmp-dependent and -independent collective cancer cell invasion. Phys. Biol. 8, 015010 (2011).
Codelli, J. A., Baskin, J. M., Agard, N. J. & Berozzi, C. R. Second-generation difluorinated cyclooctynes for copper-free click chemistry. J. Am. Chem. Soc. 130, 11486–11493 (2008).
Adzima, B. J. et al. Spatial and temporal control of the alkyne-azide cycloaddition by photoinitiated Cu(II) reduction. Nature Chem. 3, 258–261 (2011).
Hoyle, C. E. & Bowman, C. N. Thiol-ene click chemistry. Angew. Chem. Int. Ed. 49, 1540–1573 (2010).
Dondoni, A. The emergence of thiol-ene coupling as a click process for materials and bioorganic chemistry. Angew. Chem. Int. Ed. 47, 8995–8997 (2008).
Polizzotti, B. D., Fairbanks, B. D. & Anseth, K. S. Three-dimensional biochemical patterning of click-based composite hydrogels via thiolene photopolymerization. Biomacromolecules 9, 1084–1087 (2008).
DeForest, C. A., Sims, E. A. & Anseth, K. S. Peptide-functionalized click hydrogels with independently tunable mechanics and chemical functionality for 3D cell culture. Chem. Mater. 22, 4783–4790 (2010).
Killops, K. L., Campos, L. M. & Hawker, C. J. Robust, efficient, and orthogonal synthesis of dendrimers via thiol-ene ‘click’ chemistry. J. Am. Chem. Soc. 130, 5062–5064 (2008).
Fairbanks, B. D. et al. A versatile synthetic extracellular matrix mimic via thiol-norbornene photopolymerization. Adv. Mater. 21, 5005–5010 (2009).
Gupta, N. et al. A versatile approach to high-throughput microarrays using thiol-ene chemistry. Nature Chem. 2, 138–145 (2010).
Uygun, M., Tasdelen, M. A. & Yagci, Y. Influence of type of initiation on thiol-ene ‘click’ chemistry. Macromol. Chem. Phys. 211, 103–110 (2010).
Alvarez, M. et al. Single-photon and two-photon induced photocleavage for monolayers of an alkyltriethoxysilane with a photoprotected carboxylic ester. Adv. Mater. 20, 4563–4567 (2008).
Deiters, A. Principles and applications of the photochemical control of cellular processes. ChemBioChem 11, 47–53 (2010).
Holmes, C. P. Model studies for new o-nitrobenzyl photolabile linkers: substituent effects on the rates of photochemical cleavage. J. Org. Chem. 62, 2370–2380 (1997).
Ohmuro-Matsuyama, Y. & Tatsu, Y. Photocontrolled cell adhesion on a surface functionalized with a caged arginine-glycine-aspartate peptide. Angew. Chem. Int. Ed. 47, 7527–7529 (2008).
Johnson, J. A., Baskin, J. M., Bertozzi, C. R., Koberstein, J. T. & Turro, N. J. Copper-free click chemistry for the in situ crosslinking of photodegradable star polymers. Chem. Commun. 3064–3066 (2008).
Wong, D. Y., Griffin, D. R., Reed, J. & Kasko, A. M. Photodegradable hydrogels to generate positive and negative features over multiple length scales. Macromolecules 43, 2824–2831 (2010).
Kloxin, A. M., Tibbitt, M. W. & Anseth, K. S. Synthesis of photodegradable hydrogels as dynamically tunable cell culture platforms. Nature Protoc. 5, 1867–1887 (2010).
Kloxin, A. M., Benton, J. A. & Anseth, K. S. In situ elasticity modulation with dynamic substrates to direct cell phenotype. Biomaterials 31, 1–8 (2010).
Frey, M. T. & Wang, Y. L. A photo-modulatable material for probing cellular responses to substrate rigidity. Soft Matter 5, 1918–1924 (2009).
Redick, S. D., Settles, D. L., Briscoe, G. & Erickson, H. P. Defining fibronectin's cell adhesion synergy site by site-directed mutagenesis. J. Cell Biol. 149, 521–527 (2000).
Sims, E. A., DeForest, C. A. & Anseth, K. S. A mild, large-scale synthesis of 1,3-cyclooctanedione: expanding access to difluorinated cyclooctyne for copper-free click chemistry. Tetrahedron Lett. 52, 1871–1873 (2011).
Acknowledgements
The authors thank A. Kloxin and M. Tibbitt for useful discussions regarding photopatterning, C-C. Lin for advice on cell outgrowth experiments, A. Aimetti and P. Hume for communication on general experimental design, and C. Kloxin for insightful feedback on the written manuscript. Fellowship assistance to C.A.D. was awarded by the US Department of Education's Graduate Assistantships in Areas of National Need. This work was made possible by financial support from the National Science Foundation (DMR 1006711) and the Howard Hughes Medical Institute.
Author information
Authors and Affiliations
Contributions
C.A.D. and K.S.A. developed the material concept. C.A.D. and K.S.A. designed the experiments. C.A.D. carried out the experiments. C.A.D. and K.S.A. wrote the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary information
Supplementary information (PDF 2424 kb)
Rights and permissions
About this article
Cite this article
DeForest, C., Anseth, K. Cytocompatible click-based hydrogels with dynamically tunable properties through orthogonal photoconjugation and photocleavage reactions. Nature Chem 3, 925–931 (2011). https://doi.org/10.1038/nchem.1174
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nchem.1174
This article is cited by
-
Middle-out methods for spatiotemporal tissue engineering of organoids
Nature Reviews Bioengineering (2023)
-
Light-based vat-polymerization bioprinting
Nature Reviews Methods Primers (2023)
-
Topology mediates transport of nanoparticles in macromolecular networks
Nature Communications (2022)
-
Sensitizer-enhanced two-photon patterning of biomolecules in photoinstructive hydrogels
Communications Materials (2022)
-
Chemical strategies to engineer hydrogels for cell culture
Nature Reviews Chemistry (2022)