Microparticles (MPs) have activity in thrombus promotion and generation. Erythrocyte microparticles (ErMPs) have been reported to accelerate fibrinolysis in the absence of permeation. We hypothesized that shear induced ErMPs would affect fibrin structure of clots and change flow with implications for fibrinolysis.
OBJECTIVE:
To determine the effect of ErMPs on clot structure and fibrinolysis.
METHODS:
Plasma with elevated ErMPs was isolated from whole blood or from washed red blood cells (RBCs) resuspended in platelet free plasma (PFP) after high shear. Dynamic light scattering (DLS) provided size distribution of ErMPs from sheared samples and unsheared PFP controls. Clots were formed by recalcification for flow/lysis experiments and examined by confocal microscopy and SEM. Flow rates through clots and time-to-lysis were recorded. A cellular automata model showed the effect of ErMPs on fibrin polymerization and clot structure.
RESULTS:
Coverage of fibrin increased by 41% in clots formed from plasma of sheared RBCs in PFP over controls. Flow rate decreased by 46.7% under a pressure gradient of 10 mmHg/cm with reduction in time to lysis from 5.7 ± 0.7 min to 12.2 ± 1.1 min (p < 0.01). Particle size of ErMPs from sheared samples (200 nm) was comparable to endogenous microparticles.
CONCLUSIONS:
ErMPs alter the fibrin network in a thrombus and affect hydraulic permeability resulting in decelerated delivery of fibrinolytic drugs.
BadimonLMicrovesicles in atherosclerosis and angiogenesis: From bench to bedside and reverse. Front Cardiovasc Med. 2017;4:77.
2.
JeskeWPBlood cell microparticles as biomarkers of hemostatic abnormalities in patients with implanted cardiac assist devices. Biomarkers in Medicine. 2016;10(10):1095–104.
3.
WestermanMPorterJB. Red blood cell-derived microparticles: An overview. Blood Cells Molecules and Diseases. 2016;59:134–9.
4.
NascimbeneAAssociation between cell-derived microparticles and adverse events in patients with nonpulsatile Left ventricular assist devices. Journal of Heart and Lung Transplantation. 2014;33(5):470–77.
5.
HanJJAckerMAAtluriP. Left ventricular assist devices synergistic model between technology and medicine. Circulation. 2018;138(24):2841–51.
6.
AugustineDDynamic release and clearance of circulating microparticles during cardiac stress. Circulation Research. 2014;114(1):109–13.
7.
BurnierLCell-derived microparticles in haemostasis and vascular medicine. Thrombosis and Haemostasis. 2009;101(3):439–51.
8.
ZubairovaLDCirculating microparticles alter formation, structure, and properties of fibrin clots. Sci Rep. 2015;5:17611.
9.
FischerDMicroparticles from stored red blood cells enhance procoagulant and proinflammatory activity. Transfusion. 2017;57(11):2701–11.
10.
MauseSFWeberC. Microparticles protagonists of a novel communication network for intercellular information exchange. Circulation Research. 2010;107(9):1047–57.
11.
KoshiarRLErythrocyte-derived microparticles supporting activated protein c-mediated regulation of blood coagulation. PLoS One. 2014;9(8).
12.
WeiselJWLitvinovRI. Red blood cells: The forgotten player in hemostasis and thrombosis. Journal of Thrombosis and Haemostasis. 2019;17(2):271–82.
13.
NomuraSShimizuM. Clinical significance of procoagulant microparticles. J Intensive Care. 2015;3(1):2.
14.
BiroEHuman cell-derived microparticles promote thrombus formation in vivo in a tissue factor-dependent manner. J Thromb Haemost. 2003;1(12):2561–8.
15.
KalafatisMNegrescuEByzovaTPlowEF. Platelets and prothrombinPlatelet Function. In: QuinnMFitzgeraldD (eds) Humana Press; 2005.
16.
ReddyECRandML. Procoagulant phosphatidylserine-exposing platelets in vitro and in vivo. Front Cardiovasc Med. 2020;7:15.
17.
NoubouossieDFRed blood cell microvesicles activate the contact system, leading to factor IX activation via 2 independent pathways. Blood. 2020;135(10):755–65.
18.
StaessensSStructural analysis of ischemic stroke thrombi: Histological indications for therapy resistance. Haematologica. 2020;105(2):498–507.
19.
JangIKDifferential sensitivity of erythrocyte-rich and platelet-rich arterial thrombi to lysis with recombinant tissue-type plasminogen activator. A possible explanation for resistance to coronary thrombolysis. Circulation. 1989;79(4):920–8.
20.
WufsusARMaceraNENeevesKB. The hydraulic permeability of blood clots as a function of fibrin and platelet density. Biophysical Journal. 2013;104(8):1812–23.
21.
LitvinovRIThe platelet integrin alphaIIbbeta3 differentially interacts with fibrin versus fibrinogen. J Biol Chem. 2016;291(15):7858–67.
22.
ByrnesJRFactor XIIIa-dependent retention of red blood cells in clots is mediated by fibrin alpha-chain crosslinking. Blood. 2015;126(16):1940–8.
23.
TehraniSAtorvastatin has antithrombotic effects in patients with type 1 diabetes and dyslipidemia. Thrombosis Research. 2010;126(3):E225–E231.
24.
LevinGSukharevaELavrentievaA. Impact of microparticles derived from erythrocytes on fibrinolysis. Journal of Thrombosis and Thrombolysis. 2016;41(3):452–8.
25.
BerckmansRJExtracellular vesicles and coagulation in blood from healthy humans revisited. Journal of Extracellular Vesicles. 2019;8(1).
26.
SansoneRMacrovascular and microvascular function after implantation of left ventricular assist devices in end-stage heart failure: Role of microparticles. Journal of Heart and Lung Transplantation. 2015;34(7):921–32.
27.
HathcockJJ. Flow effects on coagulation and thrombosis. Arteriosclerosis Thrombosis and Vascular Biology. 2006;26(8):1729–37.
28.
DintenfassL. Effect of velocity gradient on the clotting time of blood and on the consistency of clots formed in vitro. Circ Res. 1966;18(4):349–56.
29.
NobiliMPlatelet activation due to hemodynamic shear stresses: Damage accumulation model and comparison to in vitro measurements. ASAIO J. 2008;54(1):64–72.
30.
WuJHSiddiquiKDiamondSL. Transport phenomena and clot dissolving therapy: An experimental investigation of diffusion-controlled and permeation-enhanced fibrinolysis. Thromb Haemost. 1994;72(1):105–12.
31.
BannishBEMolecular and physical mechanisms of fibrinolysis and thrombolysis from mathematical modeling and experiments. Sci Rep. 2017;7(1):6914.
32.
LeachJKPattersonEO’RearEA. Distributed intraclot thrombolysis: Mechanism of accelerated thrombolysis with encapsulated plasminogen activators. J Thromb Haemost. 2004;2(9):1548–55.
LacroixRImpact of pre-analytical parameters on the measurement of circulating microparticles: Towards standardization of protocol. J Thromb Haemost. 2012;10(3):437–46.
35.
ChenZFlow features and device-induced blood trauma in CF-VADs under a pulsatile blood flow condition: A CFD comparative study. Int J Numer Method Biomed Eng. 2018;34(2).
36.
GloverCJDynamic coagulation studies: Influence of normal and abnormal platelets on clot structure formation. Thromb Res. 1975;7(1):185–98.
37.
GloverCJRheological properties of fibrin clots. Effects of fibrinogen concentration, Factor XIII deficiency, and Factor XIII inhibition. J Lab Clin Med. 1975;86(4):644–56.
38.
SmithMABar-YamYRabinYMargolusNToffoliT. Cellular automaton simulation of polymersMRS Online Proceedings Library (OPL). Vol. 248, Materials Research Society; 1991. doi:10.1557/PROC-248-483.
39.
LawrieASThe characterization and impact of microparticles on haemostasis within fresh-frozen plasma. Vox Sang. 2008;95(3):197–204.
40.
BuerckJPProduction of erythrocyte microparticles in a sub-hemolytic environment. Journal of Artificial Organs. 2021;24(2):135–45.
41.
GershKCNagaswamiCWeiselJW. Fibrin network structure and clot mechanical properties are altered by incorporation of erythrocytes. Thromb Haemost. 2009;102(6):1169–75.
42.
HigdonJJLFordGD. Permeability of three-dimensional models of fibrous porous media. Journal of Fluid Mechanics. 1996;308:341–61.
43.
ClagueDSPhillipsRJ. A numerical calculation of the hydraulic permeability of three-dimensional disordered fibrous media. Physics of Fluids. 1997;9(6):1562–72.
44.
DiamondSLAnandS. Inner clot diffusion and permeation during fibrinolysis. Biophysical Journal. 1993;65(6):2622–43.
45.
ChopardBLattice-gas cellular automaton models for biology: From fluids to cells. Acta Biotheoretica. 2010;58(4):329–40.
46.
XuZLComputational approaches to studying thrombus development. Arteriosclerosis Thrombosis and Vascular Biology. 2011;31(3):500–5.
47.
Prieto-LangaricaAA cellular automata model of infection control on medical implants. Appl Appl Math. 2011;6(1):1–10.
48.
HugelBElevated levels of circulating procoagulant microparticles in patients with paroxysmal nocturnal hemoglobinuria and aplastic anemia. Blood. 1999;93(10):3451–6.
49.
TomkinsAJPlatelet rich clots are resistant to lysis by thrombolytic therapy in a rat model of embolic stroke. Exp Transl Stroke Med. 2015;7:2.
50.
Roka-MoiiaY. Shear-mediated platelet activation in the free flow II: Evolving mechanobiological mechanisms reveal an identifiable signature of activation and a bi-directional platelet dyscrasia with thrombotic and bleeding features. J Biomech. 2021;123:110415.
51.
LutzHUBogdanovaA. Mechanisms tagging senescent red blood cells for clearance in healthy humans. Frontiers in Physiology. 2013;4:387–402.
52.
BuerckJPA flow induced autoimmune response and accelerated senescence of red blood cells in cardiovascular devices. Scientific Reports. 2019;9(1):1–9.
