Formation and Destruction of Platelet Aggregates in Viscometric Flow

1975 ◽  
Author(s):  
H. Rieger ◽  
H. Schmid-Schönbein

Even after pseudopodia formation platelets - unlike all other known formed blood elements - remain dispersed in stasis and creeping flow and become aggregated only in the presence of a minimum amount of shearing. The “rheoaggregometer” (Rieger et al., Pflüger’s Archiv, 343, R 33, 1973) allows to measure the minimum shear rates necessary for platelet aggregation (PA), as well as the initial rate and the maximum extent of PA in citrated PRP.PA is quantified photometrically as a function of variable shear rates. The initial rate of PA steadily increases with increasing shear rates up to 460 sec-1. However, the maximal extent of PA (indicating the mechanical integrity of formed aggregates) saturates at about 35 sec-1 and then decreases because of a destruction of formed aggregates and of prevention of further PA. The aggregability of the platelets, as reflected by various degrees of shape changes, is enhanced by a drop of temperature and a rise in pH as well as by the so called aggregating agents (e.g. epinephrine 10-6 up to 10-9 M/l) : consecutively lower shear rates (lower effects of collision) are necessary to induce PA. In citrated PRP stable platelet aggregates are produced only within a defined range of shear rates. Platelet aggregability and aggregate stability are independent variables influenced by different experimental conditions.

1985 ◽  
Vol 54 (04) ◽  
pp. 857-861 ◽  
Author(s):  
Andrea Remuzzi ◽  
Lucia Raffaella Languino ◽  
Vincenzo Costantini ◽  
Vincenzo Guardabasso ◽  
Giovanni de Gartano ◽  
...  

SummaryThe adherence of human 3H-adenine-labeled platelets to rat subendothelium was quantitated using a rotating probe device. Platelet adhesion increased in relation to the rotation time, reaching a plateau value in about 4-6 min without any further increase. A non-linear fitting analysis of experimental data allowed calculations of initial rate and plateau value of platelet adhesion. Increasing the shear rates (from 35 to 150 sec-1) or the hematocrit (from 10% to 40%), both the adhesion rate and the plateau value were increased. When different platelet concentrations were used the adhesion rate and the plateau calculated increased with platelet concentration. Different plateau values were obtained in the experimental conditions considered. This suggests that the plateau was not reached for the complete occupation of the subendothelial surface by the adherent platelets. Experiments using two different vessels rotated in the same platelet suspension or, viceversa, the same vessel rotated successively in two fresh platelet suspensions, showed that the plateau was not determined by reduced platelet reactivity. Rotating the same vessel first in radiolabeled platelets, until the plateau was reached, and secondly in non labeled platelets, or viceversa, showed that the plateau was indeed a dynamic condition where the number of platelets adhering and detaching reached equilibrium. These observations suggest that the platelet adhesion to subendothelium is the final equilibrium of two platelet fluxes, one adhering to the surface and another detaching from the surface.


2001 ◽  
Vol 15 (06n07) ◽  
pp. 930-937 ◽  
Author(s):  
K. TANAKA ◽  
S. HASHIMOTO ◽  
T. TAKENOUCHI ◽  
I. SUGIMOTO ◽  
A. KUBONO ◽  
...  

The steady and transient stress responses were investigated from lower shear rates to higher shear rates at a given strength of the electric field, and the individual experimental conditions were reduced to Mason number ( M n). The electro-rheological response was found in the region with higher M n of the order of 10, and the transient response became faster as the shear rate increased. These results show that the effect of chance of collision among the polarized particles would play an important role even in the region.


1992 ◽  
Vol 67 (04) ◽  
pp. 453-457 ◽  
Author(s):  
Raelene L Kinlough-Rathbone ◽  
Marian A Packham ◽  
Dennis W Perry ◽  
J Fraser Mustard ◽  
Marco Cattaneo

SummaryThe stability of platelet aggregates is influenced by the extent of the release of granule contents; if release is extensive and aggregation is prolonged, deaggregation is difficult to achieve. The relative importance of the contributions of released substances to aggregate stability are not known, although stable thrombin-induced aggregates form in platelet-rich plasma from patients with barely detectable plasma or platelet fibrinogen, and ADP stabilizes thrombin-induced aggregates of platelets from patients with delta storage pool deficiency which otherwise deaggregate more readily than normal platelets. We degranulated platelets with thrombin (0.9 U/ml caused greater than 90% loss of delta and alpha granule contents) and recovered them as individual platelets in fresh medium. The degranulated platelets were reaggregated by thrombin (2 U/ml). To prevent continuing effects of thrombin, FPRCH2C1 was added when thrombin-induced aggregation of thrombin-degranulated platelets reached its maximum. EDTA (5 mM) or EGTA (5 mM) added at maximum aggregation did not deaggregate these platelets, indicating that the stability of these aggregates does not depend on Ca2+ in the medium. Whereas with control platelets a combination of PGE1 (10 μM) and chymotrypsin(10 U/ml) was required for deaggregation, with thrombin-degranulated platelets either PGE1 or chymo-trypsin alone caused extensive deaggregation. The rate and extent of deaggregation of thrombin-degranulated platelets by a combination of PGE1 and chymotrypsin was greater than with control platelets.Electron microscope gold immunocytochemistry using antihuman fibrinogen IgG, anti-von Willebrand factor and anti-fibronectin showed a) that fibrinogen in the vacuoles of degranulated platelets was visible at focal points of platelet contact in the aggregates, but that large areas of platelet contact had no fibrinogen detectable between them; and b) in comparison to fibrinogen, little fibronectin or von Willebrand factor (vWf) was detectable in the platelets.Since the linkages between thrombin-degranulated platelets reaggregated by thrombin can be disrupted either by raising cAMP (thus making glycoprotein IIb/IIIa unavailable) or by proteolysis, these linkages are less stable than those formed between normal platelets. It might therefore be expected that platelets that take part in thrombus formation and then recirculate are likely to form less stable thrombi than platelets that have not released their granule contents.


2008 ◽  
Vol 294 (4) ◽  
pp. H1833-H1839 ◽  
Author(s):  
S. C. Newcomer ◽  
C. L. Sauder ◽  
N. T. Kuipers ◽  
M. H. Laughlin ◽  
C. A. Ray

Shear rate is significantly lower in the superficial femoral compared with the brachial artery in the supine posture. The relative shear rates in these arteries of subjects in the upright posture (seated and/or standing) are unknown. The purpose of this investigation was to test the hypothesis that upright posture (seated and/or standing) would produce greater shear rates in the superficial femoral compared with the brachial artery. To test this hypothesis, Doppler ultrasound was used to measure mean blood velocity (MBV) and diameter in the brachial and superficial femoral arteries of 21 healthy subjects after being in the supine, seated, and standing postures for 10 min. MBV was significantly higher in the brachial compared with the superficial femoral artery during upright postures. Superficial femoral artery diameter was significantly larger than brachial artery diameter. However, posture had no significant effect on either brachial or superficial femoral artery diameter. The calculated shear rate was significantly greater in the brachial (73 ± 5, 91 ± 11, and 97 ± 13 s−1) compared with the superficial femoral (53 ± 4, 39 ± 77, and 44 ± 5 s−1) artery in the supine, seated, and standing postures, respectively. Contrary to our hypothesis, our current findings indicate that mean shear rate is lower in the superficial femoral compared with the brachial artery in the supine, seated, and standing postures. These findings of lower shear rates in the superficial femoral artery may be one mechanism for the higher propensity for atherosclerosis in the arteries of the leg than of the arm.


2020 ◽  
Vol 11 (1) ◽  
pp. 20190126 ◽  
Author(s):  
B. J. M. van Rooij ◽  
G. Závodszky ◽  
A. G. Hoekstra ◽  
D. N. Ku

The influence of the flow environment on platelet aggregation is not fully understood in high-shear thrombosis. The objective of this study is to investigate the role of a high shear rate in initial platelet aggregation. The haemodynamic conditions in a microfluidic device are studied using cell-based blood flow simulations. The results are compared with in vitro platelet aggregation experiments performed with porcine whole blood (WB) and platelet-rich-plasma (PRP). We studied whether the cell-depleted layer in combination with high shear and high platelet flux can account for the distribution of platelet aggregates. High platelet fluxes at the wall were found in silico . In WB, the platelet flux was about twice as high as in PRP. Additionally, initial platelet aggregation and occlusion were observed in vitro in the stenotic region. In PRP, the position of the occlusive thrombus was located more downstream than in WB. Furthermore, the shear rates and stresses in cell-based and continuum simulations were studied. We found that a continuum simulation is a good approximation for PRP. For WB, it cannot predict the correct values near the wall.


1985 ◽  
Vol 150 ◽  
pp. 357-380 ◽  
Author(s):  
Daniel M. Hanes ◽  
Douglas L. Inman

The rapid shearing of a mixture of cohesionless glass spheres and air or water was studied in an annular, parallel-plate shear cell designed after Savage (1978). Two types of flow were observed. In the first type of flow the entire mass of the granular material was mobilized. At high shear rates the shear and normal stresses were found to be quadratically dependent upon the mean shear rate (at constant volume concentration), in general agreement with the observations of Bagnold (1954) and Savage & Sayed (1984), and the ‘kinetic’ theory of Jenkins & Savage (1983). The stresses were found to be weakly dependent on the volume concentration up to approximately 0.5, and strongly dependent above this concentration. For flows in which water was the interstitial fluid, the ratio of the shear stress to the normal stress was slightly higher (than in air), and the stresses at lower shear rates were found to be more nearly linearly related to the shear rate. It is suggested that these effects are contributed to by the viscous dampening of grain motions by the water. The second type of flow was distinguished by the existence of an internal boundary above which the granular material deformed rapidly, but below which the granular material remained rigidly locked in place. The thickness of the shearing layer was measured to be between 5 and 15 grain diameters. The stress ratio at the bottom of the shearing layer was found to be nearly constant, suggesting the internal boundary is a consequence of the immersed weight of the shearing grains, and may be described by a Coulomb yield criterion. A scaled concentration is proposed to compare similar data obtained using different-sized materials or different apparatus. An intercomparison of the two types of flow studied, along with a comparison between the present experiments and those of Bagnold (1954) and Savage & Sayed (1984), suggests that the nature of the boundaries can have a significant effect upon the dynamics of the entire flow.


Author(s):  
C. W. Carter

This chapter is about practical uses of mathematical models to simplify the task of finding the best conditions under which to crystallize a macromolecule. The models describe a system’s response to changes in the independent variables under experimental control. Such a mathematical description is a surface, whose two-dimensional projections can be plotted, so it is usually called a ‘response surface’. Various methods have been described for navigating an unknown surface. They share important characteristics: experiments performed at different levels of the independent variables are scored quantitatively, and fitted implicitly or explicitly, to some model for system behaviour. Initially, one examines behaviour on a coarse grid, seeking approximate indications for multiple crystal forms and identifying important experimental variables. Later, individual locations on the surface are mapped in greater detail to optimize conditions. Finding ‘winning combinations’ for crystal growth can be approached successively with increasingly well-defined protocols and with greater confidence. Whether it is used explicitly or more intuitively, the idea of a response surface underlies the experimental investigation of all multivariate processes, like crystal growth, where one hopes to find a ‘best’ set of conditions. The optimization process is illustrated schematically in Figure 1. In general, there are three stages to this quantitative approach: (a) Design. One must first induce variation in some desired experimental result by changing the experimental conditions. Experiments are performed according to a plan or design. Decisions must be made concerning the experimental variables and how to sample them. (b) Experiments and scores. Each experiment provides an estimate for how the system behaves at the corresponding point in the experimental space. When these estimates are examined together as a group, patterns often appear. For example, a crystal polymorphism may occur only in restricted regions of the variable space explored by the experiment. (c) Fitting and testing models. Imposing a mathematical model onto such patterns provides a way to predict how the system will behave at points where there were no experiments. The better the predictions, the better the model. Adequate models provide accurate interpolation within the range of experimental variables originally sampled; occasionally a very good model will correctly predict behaviour outside it (1).


Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1110-1118 ◽  
Author(s):  
WH Reinhart ◽  
S Chien

Abstract The influence of the shape of the red blood cell during stomatocyte- echinocyte transformation on its deformability was studied by microsieving through pores with diameters of 2.6, 4.5, and 6.9 micron. A stomatocytic transformation was produced by chlorpromazine (0.02, 0.1, and 0.5 mmol/L) and an echinocytic transformation by sodium salicylate (7.5, 30, and 120 mmol/L). For spherostomatocytes, an increase in filtration resistance through 2.6 and 4.5 micron pores was observed, whereas for spheroechinocytes, a decrease in filtration resistance through 2.6 micron pores was found. Larger pores (6.9 micron) were not sensitive to those shape changes. The changes in deformability can be explained by the fact that the surface area of (sphero)-stomatocytes decreased, whereas that of (sphero)-echinocytes increased; the cell volume remained essentially constant. Echinocytes produced by 24-hour adenosine triphosphate depletion differed from drug- induced echinocytes: they had an increased cell volume at constant surface area and consequently an increased filtration resistance through 2.6- and 4.5-micron filter pores. Shape changes with spicule formation are therefore not a homogeneous entity, and cell geometric factors (eg, surface area and volume) must be assessed with care. The viscosity of red cell suspensions at a hematocrit level of 45% was higher for drug-induced echinocytes than discocytes or stomatocytes at all shear rates tested. We conclude that the normal discocyte represents an optimum shape for the flow in vivo since a stomatocytic transformation could impair the passage through the microcirculation (decrease in cell filterability) and an echinocytic transformation could impair the flow in larger vessels (increase in blood viscosity).


1983 ◽  
Vol 245 (2) ◽  
pp. H252-H258 ◽  
Author(s):  
T. Shiga ◽  
K. Imaizumi ◽  
N. Harada ◽  
M. Sekiya

An apparatus for determining the velocity of erythrocyte rouleaux formation was constructed, combining an inverted microscope, a transparent cone-plate viscometer, a TV image analyzer, and a computer. At lower shear rates, the overall process is the sedimentation and the rouleaux formation followed by the development of three-dimensional aggregates. The individual erythrocyte could be observed and the process was expressed by the time courses of the changes in the count and area of particles; taking the computed increment in the area/count, the rate of rouleaux formation could be estimated. The effects of shear rates, hematocrits, plasma proteins, and pH were quantified. The rate of rouleaux formation in autologous plasma increased by (1) lowering the shear rates (1.9 less than or equal to gamma less than or equal to 15 s-1),2) increasing the hematocrit (up to 0.6%), 3) adding human fibrinogen (up to 600 mg/dl) or gamma-globulin, and 4) increasing pH. The transformation to echinocytes or to stomatocytes decreased the rate of rouleaux formation. The pH effect was explained by the increase in mean corpuscular volume at lower pH rather than by the changes in the electrostatic repulsion or in the protein binding.


Blood ◽  
1990 ◽  
Vol 76 (7) ◽  
pp. 1336-1340 ◽  
Author(s):  
G Escolar ◽  
A Cases ◽  
E Bastida ◽  
M Garrido ◽  
J Lopez ◽  
...  

Abstract Uremic patients have an impaired platelet function that has been related to membrane glycoprotein (GP) abnormalities. Using a perfusion system, we have studied the interaction of normal and uremic platelets with vessel subendothelium (SE) under flow conditions. Reconstituted blood containing washed platelets, purified von Willebrand factor (vWF) (1 U/mL), and normal washed red blood cells was exposed to de- endothelialized rabbit segments for 10 minutes at two different shear rates (800 and 1,600 seconds-1). In some experiments a monoclonal antibody to the GPIIb-IIIa complex (EDU3) was added to the perfusates. With normal platelets, the percentage of the vessel covered by platelets (%CS) was 23.1% +/- 3.7% at 800 seconds-1 and 30% +/- 4.3% at 1,600 seconds-1. Platelets were observed in contact or forming monolayers on vessel SE. EDU3 inhibited the spreading of normal platelets. The %CS (11.1% +/- 3.3%) was statistically decreased (P less than .01) and most of the platelets were observed in contact with the vessel surface. These data indicate that, under flow conditions, the interaction of vWF with GPIIb-IIIa can support the spreading of normal platelets in the absence of exogenous fibrinogen. Under the same experimental conditions, the interaction of uremic platelets with SE was markedly impaired at both shear rates studied (P less than .01 v normal platelets). The presence of EDU3 did not modify the interaction of uremic platelets. These results confirm the impairment of the platelet adhesion observed in uremic patients. Furthermore, they indicate the presence of a functional defect in the interaction of vWF with GPIIb-IIIa. The fact that perfusions with normal and uremic platelets in the presence of an antibody to the GPIIb-IIIa complex did not show any differences gives indirect evidence on a functionally normal interaction vWF/GPIb in uremic patients.


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