title: Single-platelet nanomechanics measured by high-throughput cytometry.
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 scaffold. 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 bleeding and thrombotic disorders. 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.
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Myers
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Yongzhi
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Meredith E
authors.initials: ME
authors.lastname: Fay
authors.affiliation: School of Physics, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Tennenbaum
authors.affiliation: Joint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North Carolina 27695, USA.|Comparative Medicine Institute at North Carolina State University, Raleigh, North Carolina 27695, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Chester
authors.affiliation: School of Physics, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Jonas
authors.initials: J
authors.lastname: Cuadrado
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Sakurai
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Jong
authors.initials: J
authors.lastname: Baek
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Reginald
authors.initials: R
authors.lastname: Tran
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Jordan C
authors.initials: JC
authors.lastname: Ciciliano
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Byungwook
authors.initials: B
authors.lastname: Ahn
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Robert G
authors.initials: RG
authors.lastname: Mannino
authors.affiliation: Department of Pathology, Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Silvia T
authors.initials: ST
authors.lastname: Bunting
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Bennett
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Briones
authors.affiliation: Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|School of Physics, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Alberto
authors.initials: A
authors.lastname: Fernandez-Nieves
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, USA.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Smith
authors.affiliation: Joint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, North Carolina 27695, USA.|Comparative Medicine Institute at North Carolina State University, Raleigh, North Carolina 27695, USA.
authors.forename: Ashley C
authors.initials: AC
authors.lastname: Brown
authors.affiliation: George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Todd
authors.initials: T
authors.lastname: Sulchek
authors.affiliation: Department of Pediatrics, Division of Pediatric Hematology/Oncology, Aflac Cancer Center and Blood Disorders Service of Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology &Emory University, Atlanta, Georgia 30332, USA.|Winship Cancer Institute of Emory University, Atlanta, Georgia 30322, USA.|Parker H. Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Wilbur A
authors.initials: WA
authors.lastname: Lam
mesh_terms: D001777:Blood Coagulation / Q000502:physiology*; D001783:Blood Flow Velocity / Q000502:physiology*; D001792:Blood Platelets / Q000502:physiology*; D002478:Cells, Cultured; D055119:Elastic Modulus / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods*; D006244:Hardness / Q000502:physiology; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D053758:Nanoparticles / Q000737:chemistry; D015539:Platelet Activation / Q000502:physiology*; D010973:Platelet Adhesiveness / Q000502:physiology*