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Substrates with Engineered Step Changes in Rigidity Induce Traction Force Polarity and Durotaxis

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Abstract

Rigidity sensing plays a fundamental role in multiple cell functions ranging from migration, to proliferation and differentiation (Engler et al., Cell 126:677–689, 2006; Lo et al., Biophys. J. 79:144–152, 2000; Wells, Hepatology 47:1394–1400, 2008; Zoldan et al., Biomaterials 32:9612–9621, 2011). During migration, single cells have been reported to preferentially move toward more rigid regions of a substrate in a process termed durotaxis. Durotaxis could contribute to cell migration in wound healing and gastrulation, where local gradients in tissue rigidity have been described. Despite the potential importance of this phenomenon to physiology and disease, it remains unclear how rigidity guides these behaviors and the underlying cellular and molecular mechanisms. To investigate the functional role of subcellular distribution and dynamics of cellular traction forces during durotaxis, we developed a unique microfabrication strategy to generate elastomeric micropost arrays patterned with regions exhibiting two different rigidities juxtaposed next to each other. After initial cell attachment on the rigidity boundary of the micropost array, NIH 3T3 fibroblasts were observed to preferentially migrate toward the rigid region of the micropost array, indicative of durotaxis. Additionally, cells bridging two rigidities across the rigidity boundary on the micropost array developed stronger traction forces on the more rigid side of the substrate indistinguishable from forces generated by cells exclusively seeded on rigid regions of the micropost array. Together, our results highlighted the utility of step-rigidity micropost arrays to investigate the functional role of traction forces in rigidity sensing and durotaxis, suggesting that cells could sense substrate rigidity locally to induce an asymmetrical intracellular traction force distribution to contribute to durotaxis.

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Acknowledgments

We acknowledge financial support from the National Institutes of Health (EB00262, HL73305, and GM74048), the Penn Institute for Regenerative Medicine, the Nano/Bio Interface Center, and the Center for Musculoskeletal Disorders of the University of Pennsylvania. J. F. was partially funded by the American Heart Association Postdoctoral Fellowship, and R. D. was supported by a National Science Foundation Fellowship. We thank Pan Mao for assistance in scanning electron microscopy. The M.I.T. Microsystems Technology Laboratories is acknowledged for support in cleanroom fabrication.

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Author notes

  1. Ravi A. Desai

    Present address: National Institute for Medical Research and University College London, London, UK

  2. Jianping Fu

    Present address: Department of Mechanical and Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA

Authors and Affiliations

  1. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Mark T. Breckenridge, Ravi A. Desai, Michael T. Yang, Jianping Fu & Christopher S. Chen

  2. Max Planck Institute of Molecular Cell Biology and Genetics, 01307, Dresden, Germany

    Ravi A. Desai

  3. Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA

    Christopher S. Chen

  4. The Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, 02115, USA

    Christopher S. Chen

Authors
  1. Mark T. Breckenridge
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  2. Ravi A. Desai
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  3. Michael T. Yang
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  4. Jianping Fu
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Corresponding authors

Correspondence to Jianping Fu or Christopher S. Chen.

Additional information

Associate Editor Michael R. King oversaw the review of this article.

Mark T. Breckenridge and Ravi A. Desai contributed equally to this work.

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Supporting Figure 1. (a) Cross-sectional SEM images showing the negative Si master after the first DRIE step with a regular array of cylindrical, vertical holes of a uniform depth across the Si substrate (i). ii shows the negative Si master after the second photolithography where exposed photoresist on the Si wafer was completely dissolved but photoresist filling the cylindrical holes were remained in the holes. (b) Tilted top-view SEM images showing the negative step-rigidity Si master with different magnifications as indicated.

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Breckenridge, M.T., Desai, R.A., Yang, M.T. et al. Substrates with Engineered Step Changes in Rigidity Induce Traction Force Polarity and Durotaxis. Cel. Mol. Bioeng. 7, 26–34 (2014). https://doi.org/10.1007/s12195-013-0307-6

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