Abstract
Haemostasis occurs at sites of vascular injury, where flowing blood forms a clot, a dynamic and heterogeneous fibrin-based biomaterial. Paramount in the clot’s capability to stem haemorrhage are its changing mechanical properties, the major drivers of which are the contractile forces exerted by platelets against the fibrin scaffold1. However, how platelets transduce microenvironmental cues to mediate contraction and alter clot mechanics is unknown. This is clinically relevant, as overly softened and stiffened clots are associated with bleeding2 and thrombotic disorders3. Here, we report a high-throughput hydrogel-based platelet-contraction cytometer that quantifies single-platelet contraction forces in different clot microenvironments. We also show that platelets, via the Rho/ROCK pathway, synergistically couple mechanical and biochemical inputs to mediate contraction. Moreover, highly contractile platelet subpopulations present in healthy controls are conspicuously absent in a subset of patients with undiagnosed bleeding disorders, and therefore may function as a clinical diagnostic biophysical biomarker.
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Acknowledgements
The authors wish to thank A. Shaw of the Parker H. Petit Institute for Bioengineering and Bioscience at the Georgia Institute of Technology (GT); N. Anthony and the Emory University Integrated Cellular Imaging Microscopy Core of the Children’s Pediatric Research Center; and the GT Institute for Electronics and Nanotechnology (IEN) cleanroom. Financial support provided by NIH R01 (HL121264), NIH U54 (HL112309), and NSF CAREER (1150235) to W.A.L., as well as an AHA Postdoctoral Fellowship to D.R.M. are acknowledged. D.R.M. thanks Christy R. Dillon and Gabriel A. Kwong for comments and discussion.
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D.R.M. and W.A.L. conceived of and designed the platelet-contraction experiments. D.R.M., Y.Q., A.C.B., J.C.C., B.A., M.L.S., T.S. and W.A.L. designed and tested the platelet-contraction cytometer. D.R.M., M.T., D.C., J.C., Y.S., J.B., R.T., R.G.M., S.T.B., C.B., M.B. and A.F.-N. performed experiments. M.E.F. designed and wrote image analysis algorithms. D.R.M. and W.A.L. analysed data and wrote the manuscript.
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Myers, D., Qiu, Y., Fay, M. et al. Single-platelet nanomechanics measured by high-throughput cytometry. Nature Mater 16, 230–235 (2017). https://doi.org/10.1038/nmat4772
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DOI: https://doi.org/10.1038/nmat4772
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