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Thromb Haemost 2020; 120(05): 776-792
DOI: 10.1055/s-0040-1709524
Cellular Haemostasis and Platelets
Georg Thieme Verlag KG Stuttgart · New York

Platelet Activation via Shear Stress Exposure Induces a Differing Pattern of Biomarkers of Activation versus Biochemical Agonists

Yana Roka-Moiia
1   Department of Medicine, Sarver Heart Center, University of Arizona, Tucson, Arizona, United States
,
Ryan Walk
1   Department of Medicine, Sarver Heart Center, University of Arizona, Tucson, Arizona, United States
,
Daniel E. Palomares
1   Department of Medicine, Sarver Heart Center, University of Arizona, Tucson, Arizona, United States
,
Kaitlyn R. Ammann
1   Department of Medicine, Sarver Heart Center, University of Arizona, Tucson, Arizona, United States
,
Annalisa Dimasi
2   Department of Electronics, Information and Bioengineering, Politecnico di Milano, Milan, Italy
,
Joseph E. Italiano
3   Brigham and Woman's Hospital, Harvard Medical School, Boston, Massachusetts, United States
,
Jawaad Sheriff
4   Department of Biomedical Engineering, Stony Brook University, Stony Brook, New York, United States
,
Danny Bluestein
4   Department of Biomedical Engineering, Stony Brook University, Stony Brook, New York, United States
,
Marvin J. Slepian
1   Department of Medicine, Sarver Heart Center, University of Arizona, Tucson, Arizona, United States
4   Department of Biomedical Engineering, Stony Brook University, Stony Brook, New York, United States
› Author Affiliations Funding This work is supported by the National Institute of Health grants U01 EB012487 (to D.B., M.J.S., J.S.) and U01 HL131052 (to D.B., M.J.S., J.S.), and by the Arizona Center for Accelerated Biomedical Innovation of the University of Arizona (to Y.R.M., D.P., K.A., M.J.S.).
Further Information

Publication History

29 August 2019

02 March 2020

Publication Date:
05 May 2020 (online)

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Abstract

Background Implantable cardiovascular therapeutic devices, while hemodynamically effective, remain limited by thrombosis. A driver of device-associated thrombosis is shear-mediated platelet activation (SMPA). Underlying mechanisms of SMPA, as well as useful biomarkers able to detect and discriminate mechanical versus biochemical platelet activation, are poorly defined. We hypothesized that SMPA induces a differing pattern of biomarkers compared with biochemical agonists.

Methods Gel-filtered human platelets were subjected to mechanical activation via either uniform constant or dynamic shear; or to biochemical activation by adenosine diphosphate (ADP), thrombin receptor-activating peptide 6 (TRAP-6), thrombin, collagen, epinephrine, or arachidonic acid. Markers of platelet activation (P-selectin, integrin αIIbβ3 activation) and apoptosis (mitochondrial membrane potential, caspase 3 activation, and phosphatidylserine externalization [PSE]) were examined using flow cytometry. Platelet procoagulant activity was detected by chromogenic assay measuring thrombin generation. Contribution of platelet calcium flux in SMPA was tested employing calcium chelators, ethylenediaminetetraacetic acid (EDTA), and BAPTA-AM.

Results Platelet exposure to continuous shear stress, but not biochemical agonists, resulted in a dramatic increase of PSE and procoagulant activity, while no integrin αIIbβ3 activation occurred, and P-selectin levels remained barely elevated. SMPA was associated with dissipation of mitochondrial membrane potential, but no caspase 3 activation was observed. Shear-mediated PSE was significantly decreased by chelation of extracellular calcium with EDTA, while intracellular calcium depletion with BAPTA-AM had no significant effect. In contrast, biochemical agonists ADP, TRAP-6, arachidonic acid, and thrombin were potent inducers of αIIbβ3 activation and/or P-selectin exposure. This differing pattern of biomarkers seen for SMPA for continuous uniform shear was replicated in platelets exposed to dynamic shear stress via circulation through a ventricular assist device-propelled circulatory loop.

Conclusion Elevated shear stress, but not biochemical agonists, induces a differing pattern of platelet biomarkers—with enhanced PSE and thrombin generation on the platelet surface. This differential biomarker phenotype of SMPA offers the potential for early detection and discrimination from that mediated by biochemical agonists.

Keywords

shear-mediated platelet activation - shear stress - cardiovascular therapeutic device - phosphatidylserine externalization - biomarker

Authors' Contributions

Y.R.-M. designed and performed experiments, analyzed and interpreted data, and wrote the manuscript; R.W., D.E.P., and A.D. performed experiments; K.A. and J.E.I. participated in discussions and manuscript preparation; J.S. and D.B. provided acetylated prothrombin, developed the PAS assay protocol, participated in discussions, and manuscript preparation; M.J.S. designed research, interpreted data, and wrote the manuscript.


Supplementary Material

 

  • References

  • 1 Benjamin EJ, Virani SS, Callaway CW. , et al; American Heart Association Council on Epidemiology and Prevention Statistics Committee and Stroke Statistics Subcommittee. Heart disease and stroke statistics-2018 update: a report from the American Heart Association. Circulation 2018; 137 (12) e67-e492
    PubMedGoogle Scholar
  • 2 Levine A, Gupta CA, Gass A. Advanced heart failure management and transplantation. Cardiol Clin 2019; 37 (01) 105-111
    PubMedGoogle Scholar
  • 3 Singh M, Sporn ZA, Schaff HV, Pellikka PA. ACC/AHA versus ESC guidelines on prosthetic heart valve management: JACC guideline comparison. J Am Coll Cardiol 2019; 73 (13) 1707-1718
    PubMedGoogle Scholar
  • 4 Elbey MA, Palma Dalan LA, Attizzani GF. Value of MitraClip in reducing functional mitral regurgitation. US Cardiol Rev 2019; 13 (01) 30
    PubMedGoogle Scholar
  • 5 Kithcart AP, Beckman JA. ACC/AHA versus ESC guidelines for diagnosis and management of peripheral artery disease: JACC guideline comparison. J Am Coll Cardiol 2018; 72 (22) 2789-2801
    CrossrefPubMedGoogle Scholar
  • 6 Slaughter MS, Rogers JG, Milano CA. , et al; HeartMate II Investigators. Advanced heart failure treated with continuous-flow left ventricular assist device. N Engl J Med 2009; 361 (23) 2241-2251
    CrossrefPubMedGoogle Scholar
  • 7 Starling RC, Moazami N, Silvestry SC. , et al. Unexpected abrupt increase in left ventricular assist device thrombosis. N Engl J Med 2014; 370 (01) 33-40
    CrossrefPubMedGoogle Scholar
  • 8 Koliopoulou A, McKellar SH, Rondina M, Selzman CH. Bleeding and thrombosis in chronic ventricular assist device therapy: focus on platelets. Curr Opin Cardiol 2016; 31 (03) 299-307
    CrossrefPubMedGoogle Scholar
  • 9 Houël R, Mazoyer E, Boval B. , et al. Platelet activation and aggregation profile in prolonged external ventricular support. J Thorac Cardiovasc Surg 2004; 128 (02) 197-202
    CrossrefPubMedGoogle Scholar
  • 10 Slaughter MS, Sobieski II MA, Graham JD, Pappas PS, Tatooles AJ, Koenig SC. Platelet activation in heart failure patients supported by the HeartMate II ventricular assist device. Int J Artif Organs 2011; 34 (06) 461-468
    CrossrefPubMedGoogle Scholar
  • 11 Ding J, Chen Z, Niu S. , et al. Quantification of shear-induced platelet activation: high shear stresses for short exposure time. Artif Organs 2015; 39 (07) 576-583
    CrossrefPubMedGoogle Scholar
  • 12 Bluestein D, Girdhar G, Einav S, Slepian MJ. Device thrombogenicity emulation: a novel methodology for optimizing the thromboresistance of cardiovascular devices. J Biomech 2013; 46 (02) 338-344
    CrossrefPubMedGoogle Scholar
  • 13 Nobili M, Sheriff J, Morbiducci U, Redaelli A, Bluestein D. Platelet activation due to hemodynamic shear stresses: damage accumulation model and comparison to in vitro measurements. ASAIO J 2008; 54 (01) 64-72
    CrossrefPubMedGoogle Scholar
  • 14 Chow TW, Hellums JD, Moake JL, Kroll MH. Shear stress-induced von Willebrand factor binding to platelet glycoprotein Ib initiates calcium influx associated with aggregation. Blood 1992; 80 (01) 113-120
    CrossrefPubMedGoogle Scholar
  • 15 Brown III CH, Leverett LB, Lewis CW, Alfrey Jr CP, Hellums JD. Morphological, biochemical, and functional changes in human platelets subjected to shear stress. J Lab Clin Med 1975; 86 (03) 462-471
    PubMedGoogle Scholar
  • 16 Yin H, Liu J, Li Z, Berndt MC, Lowell CA, Du X. Src family tyrosine kinase Lyn mediates VWF/GPIb-IX-induced platelet activation via the cGMP signaling pathway. Blood 2008; 112 (04) 1139-1146
    PubMedGoogle Scholar
  • 17 Hellums JD, Peterson DM, Stathopoulos NA, Moake JL, Giorgio TD. Studies on the mechanisms of shear-induced platelet activation. In: Cerebral Ischemia and Hemorheology. Berlin, Heidelberg: Springer Berlin Heidelberg; 1987: 80-89
    CrossrefGoogle Scholar
  • 18 Slepian MJ, Sheriff J, Hutchinson M. , et al. Shear-mediated platelet activation in the free flow: Perspectives on the emerging spectrum of cell mechanobiological mechanisms mediating cardiovascular implant thrombosis. J Biomech 2017; 50: 20-25
    CrossrefPubMedGoogle Scholar
  • 19 Soares JS, Sheriff J, Bluestein D. A novel mathematical model of activation and sensitization of platelets subjected to dynamic stress histories. Biomech Model Mechanobiol 2013; 12 (06) 1127-1141
    CrossrefPubMedGoogle Scholar
  • 20 Girdhar G, Xenos M, Alemu Y. , et al. Device thrombogenicity emulation: a novel method for optimizing mechanical circulatory support device thromboresistance. PLoS One 2012; 7 (03) e32463
    CrossrefPubMedGoogle Scholar
  • 21 Consolo F, Sheriff J, Gorla S. , et al. High frequency components of hemodynamic shear stress profiles are a major determinant of shear-mediated platelet activation in therapeutic blood recirculating devices. Sci Rep 2017; 7 (01) 4994
    CrossrefPubMedGoogle Scholar
  • 22 Behan MWH, Storey RF. Antiplatelet therapy in cardiovascular disease. Postgrad Med J 2004; 80 (941) 155-164
    CrossrefPubMedGoogle Scholar
  • 23 Steinlechner B, Dworschak M, Birkenberg B. , et al. Platelet dysfunction in outpatients with left ventricular assist devices. Ann Thorac Surg 2009; 87 (01) 131-137
    CrossrefPubMedGoogle Scholar
  • 24 Doligalski CT, Jennings DL. Device-related thrombosis in continuous-flow left ventricular assist device support. J Pharm Pract 2016; 29 (01) 58-66
    CrossrefPubMedGoogle Scholar
  • 25 Ashbrook M, Walenga JM, Schwartz J. , et al. Left ventricular assist device-induced coagulation and platelet activation and effect of the current anticoagulant therapy regimen. Clin Appl Thromb Hemost 2013; 19 (03) 249-255
    CrossrefPubMedGoogle Scholar
  • 26 Birschmann I, Dittrich M, Eller T. , et al. Ambient hemolysis and activation of coagulation is different between HeartMate II and HeartWare left ventricular assist devices. J Heart Lung Transplant 2014; 33 (01) 80-87
    CrossrefPubMedGoogle Scholar
  • 27 Dimasi A, Roka-Moiia Y, Consolo F. , et al. Microfluidic flow-based platforms for induction and analysis of dynamic shear-mediated platelet activation-initial validation versus the standardized hemodynamic shearing device. Biomicrofluidics 2018; 12 (04) 042208
    CrossrefPubMedGoogle Scholar
  • 28 Sheriff J, Soares JS, Xenos M, Jesty J, Slepian MJ, Bluestein D. Evaluation of shear-induced platelet activation models under constant and dynamic shear stress loading conditions relevant to devices. Ann Biomed Eng 2013; 41 (06) 1279-1296
    CrossrefPubMedGoogle Scholar
  • 29 Chiu W-C, Girdhar G, Xenos M. , et al. Thromboresistance comparison of the HeartMate II ventricular assist device with the device thrombogenicity emulation- optimized HeartAssist 5 VAD. J Biomech Eng 2014; 136 (02) 021014
    CrossrefPubMedGoogle Scholar
  • 30 Schulz-Heik K, Ramachandran J, Bluestein D, Jesty J. The extent of platelet activation under shear depends on platelet count: differential expression of anionic phospholipid and factor Va. Pathophysiol Haemost Thromb 2005; 34 (06) 255-262
    CrossrefPubMedGoogle Scholar
  • 31 Clark SR, Thomas CP, Hammond VJ. , et al. Characterization of platelet aminophospholipid externalization reveals fatty acids as molecular determinants that regulate coagulation. Proc Natl Acad Sci U S A 2013; 110 (15) 5875-5880
    CrossrefPubMedGoogle Scholar
  • 32 Ilkan Z, Wright JR, Goodall AH, Gibbins JM, Jones CI, Mahaut-Smith MP. Evidence for shear-mediated Ca2+ entry through mechanosensitive cation channels in human platelets and a megakaryocytic cell line. J Biol Chem 2017; 292 (22) 9204-9217
    CrossrefPubMedGoogle Scholar
  • 33 Sheriff J, Bluestein D, Girdhar G, Jesty J. High-shear stress sensitizes platelets to subsequent low-shear conditions. Ann Biomed Eng 2010; 38 (04) 1442-1450
    CrossrefPubMedGoogle Scholar
  • 34 Schmitz G, Rothe G, Ruf A. , et al. European Working Group on Clinical Cell Analysis: consensus protocol for the flow cytometric characterisation of platelet function. Thromb Haemost 1998; 79 (05) 885-896
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Saboor M, Moinuddin M, Ilyas S. New horizons in platelets flow cytometry. Malays J Med Sci 2013; 20 (02) 62-66
    PubMedGoogle Scholar
  • 36 Gyulkhandanyan AV, Mutlu A, Allen DJ, Freedman J, Leytin V. BH3-mimetic ABT-737 induces strong mitochondrial membrane depolarization in platelets but only weakly stimulates apoptotic morphological changes, platelet shrinkage and microparticle formation. Thromb Res 2014; 133 (01) 73-79
    CrossrefPubMedGoogle Scholar
  • 37 Özgen U, Savaşan S, Buck S, Ravindranath Y. Comparison of DiOC(6)(3) uptake and annexin V labeling for quantification of apoptosis in leukemia cells and non-malignant T lymphocytes from children. Cytometry 2000; 42 (01) 74-78
    CrossrefPubMedGoogle Scholar
  • 38 Goette NP, Glembotsky AC, Lev PR. , et al. Platelet apoptosis in adult immune thrombocytopenia: insights into the mechanism of damage triggered by auto-antibodies. PLoS One 2016; 11 (08) e0160563
    CrossrefPubMedGoogle Scholar
  • 39 Jesty J, Bluestein D. Acetylated prothrombin as a substrate in the measurement of the procoagulant activity of platelets: elimination of the feedback activation of platelets by thrombin. Anal Biochem 1999; 272 (01) 64-70
    CrossrefPubMedGoogle Scholar
  • 40 Sakurai Y, Fitch-Tewfik JL, Qiu Y. , et al. Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition. Blood 2015; 126 (04) 531-538
    PubMedGoogle Scholar
  • 41 Jesty J, Yin W, Perrotta P, Bluestein D. Platelet activation in a circulating flow loop: combined effects of shear stress and exposure time. Platelets 2003; 14 (03) 143-149
    CrossrefPubMedGoogle Scholar
  • 42 Bennett JS. αIIbβ3 (GPIIb/IIIa) Structure and Function. Platelets in Thrombotic and Non-Thrombotic Disorders: Pathophysiology, Pharmacology and Therapeutics: an Update P. Gresele, N. S. Kleiman, J. A. Lopez and C. P. Page. Cham, Springer International Publishing; 2017; 99-112
    PubMedGoogle Scholar
  • 43 Topalov NN, Kotova YN, Vasil'ev SA, Panteleev MA. Identification of signal transduction pathways involved in the formation of platelet subpopulations upon activation. Br J Haematol 2012; 157 (01) 105-115
    CrossrefPubMedGoogle Scholar
  • 44 Obydennyy SI, Sveshnikova AN, Ataullakhanov FI, Panteleev MA. Dynamics of calcium spiking, mitochondrial collapse and phosphatidylserine exposure in platelet subpopulations during activation. J Thromb Haemost 2016; 14 (09) 1867-1881
    CrossrefPubMedGoogle Scholar
  • 45 Dachary-Prigent J, Freyssinet JM, Pasquet JM, Carron JC, Nurden AT. Annexin V as a probe of aminophospholipid exposure and platelet membrane vesiculation: a flow cytometry study showing a role for free sulfhydryl groups. Blood 1993; 81 (10) 2554-2565
    CrossrefPubMedGoogle Scholar
  • 46 Leytin V, Allen DJ, Mykhaylov S, Lyubimov E, Freedman J. Thrombin-triggered platelet apoptosis. J Thromb Haemost 2006; 4 (12) 2656-2663
    CrossrefPubMedGoogle Scholar
  • 47 Bonomini M, Dottori S, Amoroso L, Arduini A, Sirolli V. Increased platelet phosphatidylserine exposure and caspase activation in chronic uremia. J Thromb Haemost 2004; 2 (08) 1275-1281
    CrossrefPubMedGoogle Scholar
  • 48 Leytin V, Allen DJ, Mykhaylov S. , et al. Pathologic high shear stress induces apoptosis events in human platelets. Biochem Biophys Res Commun 2004; 320 (02) 303-310
    CrossrefPubMedGoogle Scholar
  • 49 Gyulkhandanyan AV, Mutlu A, Freedman J, Leytin V. Selective triggering of platelet apoptosis, platelet activation or both. Br J Haematol 2013; 161 (02) 245-254
    CrossrefPubMedGoogle Scholar
  • 50 Leytin V, Mykhaylov S, Allen DJ. , et al. Chemical agonists and high shear stress induce apoptosis in human platelets. Blood 2015; 104 (11) 3875
    PubMedGoogle Scholar
  • 51 Delaney MK, Liu J, Kim K. , et al. Agonist-induced platelet procoagulant activity requires shear and a Rac1-dependent signaling mechanism. Blood 2014; 124 (12) 1957-1967
    CrossrefPubMedGoogle Scholar
  • 52 Varga-Szabo D, Braun A, Nieswandt B. Calcium signaling in platelets. J Thromb Haemost 2009; 7 (07) 1057-1066
    CrossrefPubMedGoogle Scholar
  • 53 Morel O, Jesel L, Freyssinet JM, Toti F. Cellular mechanisms underlying the formation of circulating microparticles. Arterioscler Thromb Vasc Biol 2011; 31 (01) 15-26
    CrossrefPubMedGoogle Scholar
  • 54 Delaney MK, Liu J, Kim K. , et al. Agonist-induced platelet procoagulant activity requires shear and a Rac1-dependent signaling mechanism. Blood 2014; 124 (12) 1957-1967
    CrossrefPubMedGoogle Scholar
  • 55 Rivera J, Lozano ML, Navarro-Núñez L, Vicente V. Platelet receptors and signaling in the dynamics of thrombus formation. Haematologica 2009; 94 (05) 700-711
    CrossrefPubMedGoogle Scholar
  • 56 Kroll MH, Hellums JD, McIntire LV, Schafer AI, Moake JL. Platelets and shear stress. Blood 1996; 88 (05) 1525-1541
    CrossrefPubMedGoogle Scholar
  • 57 Helms CC, Marvel M, Zhao W. , et al. Mechanisms of hemolysis-associated platelet activation. J Thromb Haemost 2013; 11 (12) 2148-2154
    CrossrefPubMedGoogle Scholar
  • 58 Gremmel T, Fedrizzi S, Weigel G, Eichelberger B, Panzer S. Underlying mechanism and specific prevention of hemolysis-induced platelet activation. Platelets 2017; 28 (06) 555-559
    CrossrefPubMedGoogle Scholar
  • 59 Sakurai Y, Shima M, Giddings J. , et al. A critical role for thrombin in platelet aggregation under high shear stress. Thromb Res 2004; 113 (05) 311-318
    CrossrefPubMedGoogle Scholar
  • 60 Valerio L, Consolo F, Bluestein D. , et al. Shear-mediated platelet activation in patients implanted with continuous flow LVADs: a preliminary study utilizing the platelet activity state (PAS) assay. Proceedings of the Annual International Conference of the IEEE Engineering in Medicine and Biology Society; 2015 ;2015 November: 1255-1258
    PubMedGoogle Scholar
  • 61 Krajewski S, Krauss S, Kurz J, Neumann B, Schlensak C, Wendel HP. Real-time measurement of free thrombin: evaluation of the usability of a new thrombin assay for coagulation monitoring during extracorporeal circulation. Thromb Res 2014; 133 (03) 455-463
    CrossrefPubMedGoogle Scholar
  • 62 Yin W, Shanmugavelayudam SK, Rubenstein DA. The effect of physiologically relevant dynamic shear stress on platelet and endothelial cell activation. Thromb Res 2011; 127 (03) 235-241
    CrossrefPubMedGoogle Scholar
  • 63 Geisen U, Brehm K, Trummer G. , et al. Platelet secretion defects and acquired von Willebrand syndrome in patients with ventricular assist devices. J Am Heart Assoc 2018; 7 (02) e006519
    PubMedGoogle Scholar
  • 64 Chen Z, Mondal NK, Ding J, Gao J, Griffith BP, Wu ZJ. Shear-induced platelet receptor shedding by non-physiological high shear stress with short exposure time: glycoprotein Ibα and glycoprotein VI. Thromb Res 2015; 135 (04) 692-698
    CrossrefPubMedGoogle Scholar
  • 65 Houël R, Mazoyer E, Boval B. , et al. Platelet activation and aggregation profile in prolonged external ventricular support. J Thorac Cardiovasc Surg 2004; 128 (02) 197-202
    CrossrefPubMedGoogle Scholar
  • 66 Mondal NK, Sorensen EN, Hiivala NJ. , et al. Intraplatelet reactive oxygen species, mitochondrial damage and platelet apoptosis augment non-surgical bleeding in heart failure patients supported by continuous-flow left ventricular assist device. Platelets 2015; 26 (06) 536-544
    CrossrefPubMedGoogle Scholar
  • 67 Mondal NK, Li T, Chen Z. , et al. Mechanistic insight of platelet apoptosis leading to non-surgical bleeding among heart failure patients supported by continuous-flow left ventricular assist devices. Mol Cell Biochem 2017; 433 (1-2): 125-137
    CrossrefPubMedGoogle Scholar
  • 68 Consolo F, Sferrazza G, Motolone G. , et al. Platelet activation is a preoperative risk factor for the development of thromboembolic complications in patients with continuous-flow left ventricular assist device. Eur J Heart Fail 2018; 20 (04) 792-800
    CrossrefPubMedGoogle Scholar
  • 69 Nieswandt B, Schulte V, Zywietz A, Gratacap M-P, Offermanns S. Costimulation of Gi- and G12/G13-mediated signaling pathways induces integrin α IIbbeta 3 activation in platelets. J Biol Chem 2002; 277 (42) 39493-39498
    CrossrefPubMedGoogle Scholar
  • 70 Alonso MT, Sanchez A, Garcia-Sancho J. Arachidonic acid-induced calcium influx in human platelets. Comparison with the effect of thrombin. Biochem J 1990; 272 (02) 435-443
    CrossrefPubMedGoogle Scholar
  • 71 Rukoyatkina N, Mindukshev I, Walter U, Gambaryan S. Dual role of the p38 MAPK/cPLA2 pathway in the regulation of platelet apoptosis induced by ABT-737 and strong platelet agonists. Cell Death Dis 2013; 4 (11) e931-e939
    CrossrefPubMedGoogle Scholar
  • 72 Cao Y, Pearman AT, Zimmerman GA, McIntyre TM, Prescott SM. Intracellular unesterified arachidonic acid signals apoptosis. Proc Natl Acad Sci U S A 2000; 97 (21) 11280-11285
    CrossrefPubMedGoogle Scholar
  • 73 Penzo D, Petronilli V, Angelin A. , et al. Arachidonic acid released by phospholipase A(2) activation triggers Ca(2+)-dependent apoptosis through the mitochondrial pathway. J Biol Chem 2004; 279 (24) 25219-25225
    CrossrefPubMedGoogle Scholar
  • 74 Tonon G, Luo X, Greco NJ, Chen W, Shi Y, Jamieson GA. Weak platelet agonists and U46619 induce apoptosis-like events in platelets, in the absence of phosphatidylserine exposure. Thromb Res 2002; 107 (06) 345-350
    CrossrefPubMedGoogle Scholar
  • 75 Fujii T, Sakata A, Nishimura S, Eto K, Nagata S. TMEM16F is required for phosphatidylserine exposure and microparticle release in activated mouse platelets. Proc Natl Acad Sci U S A 2015; 112 (41) 12800-12805
    CrossrefPubMedGoogle Scholar
  • 76 Yang H, Kim A, David T. , et al. TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation. Cell 2012; 151 (01) 111-122
    CrossrefPubMedGoogle Scholar
  • 77 Baig AA, Haining EJ, Geuss E. , et al. TMEM16F-mediated platelet membrane phospholipid scrambling is critical for hemostasis and thrombosis but not thromboinflammation in mice-brief report. Arterioscler Thromb Vasc Biol 2016; 36 (11) 2152-2157
    CrossrefPubMedGoogle Scholar
  • 78 Kholmukhamedov A, An A, Liu F, Janecke R, Jobe SM. Calcium independent events trigger VWF-dependent shear-induced procoagulant platelet formation. Blood 2016 128(22): http://www.bloodjournal.org/content/128/22/1354?sso-checked=true&utm_source=TrendMD&utm_medium=cpc&utm_campaign=Blood_TrendMD_0 . Accessed January 16, 2018
    PubMedGoogle Scholar
  • 79 Sweedo A, Scott Saavedra S, Sheriff J, Bluestein D, Slepian MJ. Platelet membrane fluidity: a mechanistic component of shear- mediated platelet activation. ASAIO J 2018; 64: 40
    CrossrefPubMedGoogle Scholar
  • 80 Lu Q, Malinauskas RA. Comparison of two platelet activation markers using flow cytometry after in vitro shear stress exposure of whole human blood. Artif Organs 2011; 35 (02) 137-144
    PubMedGoogle Scholar