Local nascent protein deposition and remodelling guide mesenchymal stromal cell mechanosensing and fate in three-dimensional hydrogels

Abstract

Hydrogels serve as valuable tools for studying cell–extracellular matrix interactions in three-dimensional environments that recapitulate aspects of native extracellular matrix. However, the impact of early protein deposition on cell behaviour within hydrogels has largely been overlooked. Using a bio-orthogonal labelling technique, we visualized nascent proteins within a day of culture across a range of hydrogels. In two engineered hydrogels of interest in three-dimensional mechanobiology studies—proteolytically degradable covalently crosslinked hyaluronic acid and dynamic viscoelastic hyaluronic acid hydrogels—mesenchymal stromal cell spreading, YAP/TAZ nuclear translocation and osteogenic differentiation were observed with culture. However, inhibition of cellular adhesion to nascent proteins or reduction in nascent protein remodelling reduced mesenchymal stromal cell spreading and nuclear translocation of YAP/TAZ, resulting in a shift towards adipogenic differentiation. Our findings emphasize the role of nascent proteins in the cellular perception of engineered materials and have implications for in vitro cell signalling studies and application to tissue repair.

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Fig. 1: Nascent protein deposition by encapsulated hMSCs occurs early, independent of hydrogel type.
Fig. 2: Nascent ECM proteins create an adhesive layer at the cell–hydrogel interface.
Fig. 3: Adhesion to nascent proteins controls hMSC mechanosensing in degradable hydrogels.
Fig. 4: Dynamic hydrogel composition modulates viscoelastic properties and cell spreading.
Fig. 5: Nascent protein remodelling is required for cell spreading and osteogenesis in dynamic hydrogels.
Fig. 6: Nascent protein adhesion and remodelling enhance cell spreading in degradable/dynamic hydrogels.

Data availability

All the data generated or analysed during this study are included within this article and its Supplementary Information. Additional information is available from the corresponding author on request.

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Acknowledgements

This work was supported by the Swiss National Foundation through an SNF Early Postdoc Mobility Fellowship (to C.L.), the National Science Foundation (DMR award 1610525, the Center for Engineering MechanoBiology CMMI: 15-48571) and the National Institutes of Health (R01 EB008722). We are grateful for help from the Penn EMRL Electron Microscopy Core for TEM and the Penn CDB Microscopy Core Facility for TFM, and would like to thank D. Seliktar for providing the PEG-DA, and A. Garcia, M. Davidson, R. Daniels, B. Cosgrove and M. D’Este for helpful conversations.

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C.L., R.L.M. and J.A.B. conceived the ideas and designed the experiments. C.L. conducted the experiments and analysed the data. C.L., R.L.M. and J.A.B. interpreted the data and wrote the manuscript.

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Correspondence to Jason A. Burdick.

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Loebel, C., Mauck, R.L. & Burdick, J.A. Local nascent protein deposition and remodelling guide mesenchymal stromal cell mechanosensing and fate in three-dimensional hydrogels. Nat. Mater. 18, 883–891 (2019). https://doi.org/10.1038/s41563-019-0307-6

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