Fibrin network structure and clot mechanical properties are altered by incorporation of erythrocytes

 

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Summary

Although many in vitro fibrin studies are performed with plasma, in vivo clots and thrombi contain erythrocytes, or red blood cells (RBCs).To determine the effects of RBCs on fibrin clot structure and mechanical properties, we compared plasma clots without RBCs to those prepared with low (2 vol%), intermediate (5-10 vol%), or high (≥20 vol%) numbers of RBCs. By confocal microscopy, we found that low RBC concentrations had little effect on clot structure. Intermediate RBC concentrations caused heterogeneity in the fiber network with pockets of densely packed fibers alongside regions with few fibers. With high levels of RBCs, fibers arranged more uniformly but loosely around the cells. Scanning electron micrographs demonstrated an uneven distribution of RBCs throughout the clot and a significant increase in fiber diameter upon RBC incorporation. While permeability was not affected by RBC addition, at 20% or higher RBCs, the ratio of viscous modulus (G′′) to elastic modulus (G′) increased significantly over that of a clot without any RBCs. RBCs triggered variability in the fibrin network structure, individual fiber characteristics, and overall clot viscoelasticity compared to the absence of cells. These results are important for understanding in vivo clots and thrombi.

• References

• 1 Wohner N. Role of cellular elements in thrombus formation and dissolution. Cardiovasc Hematol Agents Med Chem 2008; 06: 224-228.
  CrossrefPubMedGoogle Scholar
• 2 Zannad F, Stoltz JF. Blood rheology in arterial hypertension. J Hypertens Suppl 1992; 10: S69-78.
  PubMedGoogle Scholar
• 3 Schmid-Schonbein H, Wells R, Goldstone J. Influence of deformability of human red cells upon blood viscosity. Circ Res 1969; 25: 131-143.
  CrossrefPubMedGoogle Scholar
• 4 Landolfi R, Rocca B, Patrono C. Bleeding and thrombosis in myeloproliferative disorders: mechanisms and treatment. Crit Rev Oncol Hematol 1995; 20: 203-222.
  CrossrefPubMedGoogle Scholar
• 5 Goldsmith HL. Red cell motions and wall interactions in tube flow. Fed Proc 1971; 30: 1578-1590.
  PubMedGoogle Scholar
• 6 Goldsmith HL, Bell DN, Braovac S. et al. Physical and chemical effects of red cells in the shear-induced aggregation of human platelets. Biophys J 1995; 69: 1584-1595.
  CrossrefPubMedGoogle Scholar
• 7 Hellem AJ, Borchgrevink CF, Ames SB. The role of red cells in haemostasis: the relation between haematocrit, bleeding time and platelet adhesiveness. Br J Haematol 1961; 07: 42-50.
  CrossrefPubMedGoogle Scholar
• 8 Reinhart WH, Zehnder L, Schulzki T. Stored erythrocytes have less capacity than normal erythrocytes to support primary haemostasis. Thromb Haemost 2009; 101: 720-723.
  Article in Thieme ConnectPubMedGoogle Scholar
• 9 Zwaal RF, Schroit AJ. Pathophysiologic implications of membrane phospholipid asymmetry in blood cells. Blood 1997; 89: 1121-1132.
  PubMedGoogle Scholar
• 10 Ataga KI, Orringer EP. Hypercoagulability in sickle cell disease: a curious paradox. Am J Med 2003; 115: 721-728.
  CrossrefPubMedGoogle Scholar
• 11 Levi M, Ten Cate H. Disseminated intravascular coagulation. N Engl J Med 1999; 341: 586-592.
  CrossrefPubMedGoogle Scholar
• 12 Marder VJ, Chute DJ, Starkman S. et al. Analysis of thrombi retrieved from cerebral arteries of patients with acute ischemic stroke. Stroke 2006; 37: 2086-2093.
  CrossrefPubMedGoogle Scholar
• 13 Carr Jr ME, Hardin CL. Fibrin has larger pores when formed in the presence of erythrocytes. Am J Physiol 1987; 253: H1069-1073.
  PubMedGoogle Scholar
• 14 van Gelder JM, Nair CH, Dhall DP. The significance of red cell surface area to the permeability of fibrin network. Biorheology 1994; 31: 259-275.
  CrossrefPubMedGoogle Scholar
• 15 Weisel JW, Nagaswami C. Computer modeling of fibrin polymerization kinetics correlated with electron microscope and turbidity observations: clot structure and assembly are kinetically controlled. Biophys J 1992; 63: 111-128.
  CrossrefPubMedGoogle Scholar
• 16 Marchi R, Lopez Ramirez Y, Nagaswami C. et al. Haemostatic changes related to fibrin formation and fibrinolysis during the first trimester in normal pregnancy and in recurrent miscarriage. Thromb Haemost 2007; 97: 552-557.
  Article in Thieme ConnectPubMedGoogle Scholar
• 17 Ryan EA, Mockros LF, Weisel JW. et al. Structural origins of fibrin clot rheology. Biophys J 1999; 77: 2813-2826.
  CrossrefPubMedGoogle Scholar
• 18 Banerjee R, Nageshwari K, Puniyani RR. The diagnostic relevance of red cell rigidity. Clin Hemorheol Microcirc 1998; 19: 21-24.
  PubMedGoogle Scholar
• 19 Dobbe JG, Hardeman MR, Streekstra GJ. et al. Analyzing red blood cell-deformability distributions. Blood Cells Mol Dis 2002; 28: 373-384.
  CrossrefPubMedGoogle Scholar
• 20 Evans EA, La Celle PL. Intrinsic material properties of the erythrocyte membrane indicated by mechanical analysis of deformation. Blood 1975; 45: 29-43.
  PubMedGoogle Scholar
• 21 Yoon YZ, Kotar J, Yoon G. et al. The nonlinear mechanical response of the red blood cell. Phys Biol 2008; 05: 36007.
  CrossrefPubMedGoogle Scholar
• 22 Mohandas N, Gallagher PG. Red cell membrane: past, present, and future. Blood 2008; 112: 3939-3948.
  CrossrefPubMedGoogle Scholar
• 23 Collet JP, Allali Y, Lesty C. et al. Altered fibrin architecture is associated with hypofibrinolysis and premature coronary atherothrombosis. Arterioscler Thromb Vasc Biol 2006; 26: 2567-2573.
  CrossrefPubMedGoogle Scholar
• 24 Sjoland JA, Sidelmann JJ, Brabrand M. et al. Fibrin clot structure in patients with end-stage renal disease. Thromb Haemost 2007; 98: 339-345.
  Article in Thieme ConnectPubMedGoogle Scholar