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Cell-mediated fibre recruitment drives extracellular matrix mechanosensing in engineered fibrillar microenvironments

Abstract

To investigate how cells sense stiffness in settings structurally similar to native extracellular matrices, we designed a synthetic fibrous material with tunable mechanics and user-defined architecture. In contrast to flat hydrogel surfaces, these fibrous materials recapitulated cell–matrix interactions observed with collagen matrices including stellate cell morphologies, cell-mediated realignment of fibres, and bulk contraction of the material. Increasing the stiffness of flat hydrogel surfaces induced mesenchymal stem cell spreading and proliferation; however, increasing fibre stiffness instead suppressed spreading and proliferation for certain network architectures. Lower fibre stiffness permitted active cellular forces to recruit nearby fibres, dynamically increasing ligand density at the cell surface and promoting the formation of focal adhesions and related signalling. These studies demonstrate a departure from the well-described relationship between material stiffness and spreading established with hydrogel surfaces, and introduce fibre recruitment as a previously undescribed mechanism by which cells probe and respond to mechanics in fibrillar matrices.

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Figure 1: A new approach to engineering fibrillar microenvironments with tunable mechanical and architectural features.
Figure 2: Synthetic fibre networks induce similar topographical and mechanical interactions with cells to collagen matrices at multiple length scales.
Figure 3: Increasing fibre stiffness suppresses cell spreading and proliferation.
Figure 4: Lower fibre and network stiffness enables cell-mediated reorganization of the material and clustering of adhesive ligands local to the cell.
Figure 5: Fibrillar ECM remodelling promotes FA formation and FAK phosphorylation to increase proliferation.

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Acknowledgements

This work was supported in part by grants from the National Institutes of Health (grant numbers EB000262, EB001046, HL115553, GM74048, AR056624) and Center for Engineering Cells and Regeneration of the University of Pennsylvania. B.M.B. acknowledges financial support from a Ruth L. Kirschstein National Research Service Award (EB014691) and NIH Pathway to Independence Award (HL124322). We would like to thank N. Wang for helpful discussions. We would also like to acknowledge support from the Penn Regional Nanotechnology Facility and the BME Core Facilities at Boston University.

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B.M.B., B.T., J.A.B. and C.S.C. designed the materials. B.M.B., B.T. and C.S.C. designed the experiments. B.M.B., B.T., W.Y.W., M.S.S. and I.L.K. conducted experiments and analysed data. V.B.S. helped with analysis and interpretation of mechanical testing data. B.M.B., B.T. and C.S.C. wrote the manuscript.

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Correspondence to Brendon M. Baker or Christopher S. Chen.

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Baker, B., Trappmann, B., Wang, W. et al. Cell-mediated fibre recruitment drives extracellular matrix mechanosensing in engineered fibrillar microenvironments. Nature Mater 14, 1262–1268 (2015). https://doi.org/10.1038/nmat4444

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