Article | Published:

Hydrogels with tunable stress relaxation regulate stem cell fate and activity

Nature Materials volume 15, pages 326334 (2016) | Download Citation

Abstract

Natural extracellular matrices (ECMs) are viscoelastic and exhibit stress relaxation. However, hydrogels used as synthetic ECMs for three-dimensional (3D) culture are typically elastic. Here, we report a materials approach to tune the rate of stress relaxation of hydrogels for 3D culture, independently of the hydrogel’s initial elastic modulus, degradation, and cell-adhesion-ligand density. We find that cell spreading, proliferation, and osteogenic differentiation of mesenchymal stem cells (MSCs) are all enhanced in cells cultured in gels with faster relaxation. Strikingly, MSCs form a mineralized, collagen-1-rich matrix similar to bone in rapidly relaxing hydrogels with an initial elastic modulus of 17 kPa. We also show that the effects of stress relaxation are mediated by adhesion-ligand binding, actomyosin contractility and mechanical clustering of adhesion ligands. Our findings highlight stress relaxation as a key characteristic of cell–ECM interactions and as an important design parameter of biomaterials for cell culture.

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Acknowledgements

The authors acknowledge the help of S. Koshy, M. Mehta, C. Verbeke, X. Zhao (now at MIT), and other members of the Mooney lab. The authors also thank the Weitz lab for use of a rheometer, O. Uzun for help with GPC, S. Reinke (Berlin-Brandenburg Center for Regenerative Therapies) for providing the human bone haematoma samples, and D. Wulsten and S. Reinke for the support in bone fracture haematoma testing. This work was supported by an NIH Grant to D.J.M. (R01 DE013033), an NIH F32 grant to O.C. (CA153802), an Einstein Visiting Fellowship for D.J.M., funding of the Einstein Foundation Berlin through the Charité—Universitätsmedizin Berlin, Berlin-Brandenburg School for Regenerative Therapies GSC 203, ZonMW-VICI grant 918.11.635 (The Netherlands) for D.K., and Harvard MRSEC for D.J.M. (DMR-1420570). This work was performed in part at the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Infrastructure Network (NNIN).

Author information

Author notes

    • Ovijit Chaudhuri
    •  & Luo Gu

    These authors contributed equally to this work.

Affiliations

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

    • Ovijit Chaudhuri
    • , Luo Gu
    • , Darinka Klumpers
    • , Max Darnell
    • , Sidi A. Bencherif
    • , Nathaniel Huebsch
    •  & David J. Mooney
  2. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts 02138, USA

    • Ovijit Chaudhuri
    • , Luo Gu
    • , Darinka Klumpers
    • , Max Darnell
    • , Sidi A. Bencherif
    • , James C. Weaver
    • , Evi Lippens
    •  & David J. Mooney
  3. Department of Mechanical Engineering, Stanford University, Stanford, California 94305, USA

    • Ovijit Chaudhuri
    •  & Hong-pyo Lee
  4. Department of Orthopedic Surgery, Research Institute MOVE, VU University Medical Center, 1081 HV Amsterdam, The Netherlands

    • Darinka Klumpers
  5. Gladstone Institute of Cardiovascular Disease, San Francisco, California 94158, USA

    • Nathaniel Huebsch
  6. Julius Wolff Institute, Charité—Universitätsmedizin Berlin and Berlin-Brandenburg Center for Regenerative Therapies, 13353 Berlin, Germany

    • Evi Lippens
    •  & Georg N. Duda

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Contributions

O.C., L.G., D.K., M.D., N.H. and D.J.M. designed the experiments. O.C. and L.G. conducted most of the experiments. D.K. helped with experiments involving the MSCs. S.A.B. helped with alginate characterization. J.C.W. helped with EDS experiments and analysis. H.-p.L. assisted with mechanical characterization. E.L. and G.N.D. carried out fracture haematoma measurement. O.C. and L.G. analysed the data. O.C., L.G., D.K. and D.J.M. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David J. Mooney.

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DOI

https://doi.org/10.1038/nmat4489

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