A programmable, computer-controlled cone-plate viscometer for the application of pulsatile shear stress to platelet suspensions

Described is a special purpose cone-plate viscometer that is capable of acceleration or deceleration through a step change in speed in less than 0.7s. The speed of the rotating cone is controlled by a microcomputer which can be programmed to generate speed vs time ramp functions of variable slope. Prior calibration of motor power required to shear Newtonian fluids of known viscosity at various speeds provides the basis for determination of apparent suspension viscosity and enables the viscometer automatically to compensate for changing sample viscosity during shear. The viscometer was used to carry out a series of preliminary studies in which platelet-rich plasma (PRP) was subjected to continuous and pulsatile shear stress at 37 degrees C. Shear-induced platelet aggregation (SIPAG) was significantly greater in response to pulsatile versus continuous shearing except at the lowest applied stress (10 dyn/cm2). Increases ranged from about 40 percent at a stress amplitude of 25 dyn/cm2 to nearly 55 percent at dyn/cm2. This increasing trend with stress amplitude might be interpreted as a positive correlation between SIPAG and the loading rate. Dense granule release, as indicated by serotonin release, was dependent on both stress amplitude and number of pulses even at the higher stress where SIPAG was independent of pulse number.     

Post-stenotic core flow behavior in pulsatile flow and its effects on wall shear stress

Arteries of several species, including man, tend to adjust their diameters such that the mean wall shear stress is in the range of 10-20 dynes cm-2. Additionally, intimal thickening in the human carotid bifurcation correlates well with the reciprocal of wall shear stress as determined in model studies. The correlation indicates that wherever the local mean wall shear stress exceeds approximately 10 dynes cm-2, the artery tends to be spared from intimal thickening. However, it is not known whether mean shear stress, i.e. the time-averaged value, or the instantaneous shear stress is the appropriate correlative variable. Each of these variables suggests different mechanisms for the reaction of the artery wall to its hemodynamic environment. It is therefore important to devise means by which the effects of mean shear and pulsatile shear can be separated in the study of atherogenesis. The present investigation examines the post-stenotic flow field in Plexiglas models under pulsatile conditions approximating those in the aortas of the cynomolgus monkey, an animal often employed in atherogenesis research. Behavior of the core flow and its effects on wall shear stress are studied for stenoses of 75 and 90% area reductions using laser velocimetry. The results show that the post-stenotic field contains regions in which the mean wall shear stress is low, but the pulsatile excursions are large.(ABSTRACT TRUNCATED AT 250 WORDS)     

Breaking symmetry in non-planar bifurcations: distribution of flow and wall shear stress

Non-planarity in blood vessels is known to influence arterial flows and wall shear stress. To gain insight, computational fluid dynamics (CFD) has been used to investigate effects of curvature and out-of-plane geometry on the distribution of fluid flows and wall shear stresses in a hypothetical non-planar bifurcation. Three-dimensional Navier-Stokes equations for a steady state Newtonian fluid were solved numerically using a finite element method. Non-planarity in one of the two daughter vessels is found to deflect flow from the inner wall of the vessel to the outer wall and to cause changes in the distribution of wall shear stresses. Results from this study agree to experimental observations and CFD simulations in the literature, and support the view that non-planarity in blood vessels is a factor with important haemodynamic significance and may play a key role in vascular biology and pathophysiology.     

Three-dimensional numerical analysis of pulsatile flow and wall shear stress in the carotid artery bifurcation.

To analyse the pulsatile flow field and the mechanical stresses in a three-dimensional carotid artery bifurcation model, computer simulation is applied. The approximation of the Navier-Stokes equations uses a pressure correction finite element method. Numerical results are presented for axial and secondary flow velocity and wall shear stresses with special emphasis on the fluid dynamics in the carotid sinus. This region is of major interest because it is affected preferentially by lesions. Detailed local flow studies as carried out here should lead to a further insight into the mechanisms of atherogenesis. The flow conditions used in the study were chosen according to Ku et al. (Arteriosclerosis 5, 293-302, 1985). The results of this numerical analysis agree in the essential features with their experimental results.     

Regulation of flow and wall shear stress in arteriolar networks of the hamster cheek pouch.

Our purpose was to define arteriolar network hemodynamics during moderate increases in interstitial adenosine or nitric oxide in the hamster (n = 34, pentobarbital sodium 70 mg/kg) cheek pouch tissue. The network consists of a feed arteriole (approximately 12-microm diameter, approximately 800-microm length) with three to six branches. Observations of diameter, red blood cell flux, and velocity were obtained at the feed before the branch and within the branch. A comparison of baseline with suffused adenosine or sodium nitroprusside (SNP) 10(-9) to 10(-5) M showed the following. First, diameter change was heterogeneous by agonist, did not reflect the expected dilatory response, and was related to location within the network. With adenosine, upstream branch points constricted and those downstream dilated, even at 10(-5) M. With SNP, upstream branch points dilated, whereas those downstream constricted. Second, with adenosine, changes in diameter, flux, and velocity together resulted in no change in wall shear stress until 10(-5) M. Wall shear stress was not maintained at a constant level with Nomega-nitro-L-arginine (10(-5) M), suggesting a role for flow-dependent diameter changes with adenosine. With SNP, diameter change correlated with the baseline (before SNP) shear stress conditions.     

Tetrahedral vs. polyhedral mesh size evaluation on flow velocity and wall shear stress for cerebral hemodynamic simulation

Haemodynamic factors, in particular wall shear stresses (WSSs) may have significant impact on growth and rupture of cerebral aneurysms. Without a means to measure WSS reliably in vivo, computational fluid dynamic (CFD) simulations are frequently employed to visualise and quantify blood flow from patient-specific computational models. With increasing interest in integrating these CFD simulations into pretreatment planning, a better understanding of the validity of the calculations in respect to computation parameters such as volume element type, mesh size and mesh composition is needed. In this study, CFD results for the two most common aneurysm types (saccular and terminal) are compared for polyhedral- vs. tetrahedral-based meshes and discussed regarding future clinical applications. For this purpose, a set of models were constructed for each aneurysm with spatially varying surface and volume mesh configurations (mesh size range: 5119-258, 481 volume elements). WSS distribution on the model wall and point-based velocity measurements were compared for each configuration model. Our results indicate a benefit of polyhedral meshes in respect to convergence speed and more homogeneous WSS patterns. Computational variations of WSS values and blood velocities are between 0.84 and 6.3% from the most simple mesh (tetrahedral elements only) and the most advanced mesh design investigated (polyhedral mesh with boundary layer).     

Volume flow and wall shear stress quantification in the human conjunctival capillaries and post-capillary venules in vivo

Understanding the mathematical relationships of volume blood flow and wall shear stress with respect to microvessel diameter is necessary for the study of vascular design. Here, for the first time, volume flow and wall shear stress were quantified from axial red blood cell velocity measurements in 104 conjunctival microvessels of 17 normal human volunteers. Measurements were taken with a slit lamp based imaging system from the post capillary side of the bulbar conjunctiva in microvessel diameters ranging from 4 to 24 micrometers. The variation of the velocity profile with diameter was taken into account by using a profile factor function. Volume flow ranged from 5 to 462 pl/s with a mean value of 102 pl/s and gave a second power law best fitting line (r=0.97) deviating significantly from the third power law relation with diameter. The estimated wall shear stress declined hyperbolically (r=0.93) from a maximum of 9.55 N/m(2) at the smallest capillaries, down to a minimum of 0.28 N/m(2) at the higher diameter post capillary venules. The mean wall shear stress value for all microvessels was 1.54 N/m(2).     

Velocity and wall shear stress patterns in the human right coronary artery.

Blood flow dynamics in the human right coronary artery have not been adequately quantified despite the clinical significance of coronary atherosclerosis. In this study, a technique was developed to construct a rigid flow model from a cast of a human right coronary artery. A laser photochromic method was used to characterize the velocity and wall shear stress patterns. The flow conditions include steady flow at Reynolds numbers of 500 and 1000 as well as unsteady flow with Womersley parameter and peak Reynolds number of 1.82 and 750, respectively. Characterization of the three-dimensional geometry of the artery revealed that the largest spatial variation in curvature occurred within the almost branch-free proximal region, with the greatest curvature existing along the acute margin of the heart. In the proximal segment, high shear stresses were observed on the outer wall and lower, but not negative, stresses along the inner wall. Low shear stress on the inner wall may be related to the preferential localization of atherosclerosis in the proximal segment of the right coronary artery. However, it is possible that the large difference between the outer and inner wall shear stresses may also be involved.     

Effect of wall shear rate on biofilm deposition and grazing in drinking water flow chambers

The effect of four-wall shear rates (34.9, 74.8, 142.5, and 194.5 s(-1)) on bacterial deposition on glass slides in drinking water flow chambers was studied. Biofilm image acquisition was performed over a 50-day period. Bacterial accumulation and surface coverage curves were obtained. Microscopic observations allowed us to obtain information about the dynamics and spatial distribution of the biofilm. During the first stage of biofilm formation (210-518 h), bacterial accumulation was a function of the wall shear rate: the higher the wall shear rate, the faster the bacterial deposition (1.1 and 1.9 x 10(4) bacterial cells . cm(-2) for wall shear rates of 34.9 and 142.5 s(-1), respectively). A new similarity relationship characteristic of a non-dimensional time and function of the wall shear rate was proposed to describe initial bacterial deposition. After 50 days of exposure to drinking water, surface coverage was more or less identical under the entire wall shear rates (7.44 +/- 0.9%), suggesting that biofilm bacterial density cannot be controlled using hydrodynamics. However, the spatial distribution of the biofilm was clearly different. Under low wall shear rate, aggregates were composed of bacterial cells able to "vibrate" independently on the surface, whereas, under a high wall shear rate, aggregates were more cohesive. Therefore, susceptibility to the hydraulic discontinuities occurring in drinking water system may not be similar. In all the flow chambers, significant decreases in bacterial biomass (up to 77%) were associated with the presence of amoebae. This grazing preferentially targeted small, isolated cells.     

Effects of wall shear stress and fluid recirculation on the localization of circulating monocytes in a three-dimensional flow model

There is a correlation between the location of early atherosclerotic lesions and the hemodynamic characteristics at those sites. Circulating monocytes are key cells in the pathogenesis of atherosclerotic plaques and localize at sites of atherogenesis. The hypothesis that the distribution of monocyte adhesion to the vascular wall is determined in part by hemodynamic factors was addressed by studying monocyte adhesion in an in vitro flow model in the absence of any biological activity in the model wall.

Suspensions of U937 cells were perfused (Re = 200) through an axisymmetric silicone flow model with a stenosis followed by a reverse step. The model provided spatially varying wall shear stress, flow separation and reattachment, and a three-dimensional flow pattern. The cell rolling velocity and adhesion rates were determined by analysis of videomicrographs. Wall shear stress was obtained by numerical solution of the equations of fluid motion. Cell adhesion patterns were also studied in the presence of chemotactic peptide gradients.

The cell rolling velocity varied linearly with wall shear stress. The adhesion rate tended to decrease with increasing local wall shear stress, but was also affected by the radial component of velocity and the dynamics of the recirculation region and flow reattachment. Adhesion was increased in the vicinity of chemotactic peptide sources downstream of the expansion site. Results with human monocytes were qualitatively similar to the U937 experiments.

Differences in the adhesion rates of U937 cells occurring solely as a function of the fluid dynamic properties of the flow field were clearly demonstrated in the absence of any biological activity in the model wall.     

Combined effects of pulsatile flow and dynamic curvature on wall shear stress in a coronary artery bifurcation model

A three-dimensional model with simplified geometry for the branched coronary artery is presented. The bifurcation is defined by an analytical intersection of two cylindrical tubes lying on a sphere that represents an idealized heart surface. The model takes into account the repetitive variation of curvature and motion to which the vessel is subject during each cardiac cycle, and also includes the phase difference between arterial motion and blood flowrate, which may be nonzero for patients with pathologies such as aortic regurgitation. An arbitrary Lagrangian Eulerian (ALE) formulation of the unsteady, incompressible, three-dimensional Navier-Stokes equations is employed to solve for the flow field, and numerical simulations are performed using the spectral/hp element method. The results indicate that the combined effect of pulsatile inflow and dynamic geometry depends strongly on the aforementioned phase difference. Specifically, the main findings of this work show that the time-variation of flowrate ratio between the two branches is minimal (less than 5%) for the simulation with phase difference angle equal to 90 degrees, and maximal (51%) for 270 degrees. In two flow pulsatile simulation cases for fixed geometry and dynamic geometry with phase angle 270 degrees, there is a local minimum of the normalized wall shear rate amplitude in the vicinity of the bifurcation, while in other simulations a local maximum is observed.     

In-vivo coronary flow profiling based on biplane angiograms: influence of geometric simplifications on the three-dimensional reconstruction and wall shear stress calculation

Background  

Clinical studies suggest that local wall shear stress (WSS) patterns modulate the site and the progression of atherosclerotic lesions. Computational fluid dynamics (CFD) methods based on in-vivo three-dimensional vessel reconstructions have recently been shown to provide prognostically relevant WSS data. This approach is, however, complex and time-consuming. Methodological simplifications are desirable in porting this approach from bench to bedside. The impact of such simplifications on the accuracy of geometry and wall shear stress calculations has to be investigated.     

Oscillatory fluid flow induces the osteogenic lineage commitment of mesenchymal stem cells: The effect of shear stress magnitude,frequency, and duration

A potent regulator of bone anabolism is physical loading. However, it is currently unclear whether physical stimuli such as fluid shear within the marrow cavity is sufficient to directly drive the osteogenic lineage commitment of resident mesenchymal stem cells (MSC). Therefore, the objective of the study is to employ a systematic analysis of oscillatory fluid flow (OFF) parameters predicted to occur in vivo on early MSC osteogenic responses and late stage lineage commitment. MSCs were exposed to OFF of 1 Pa, 2 Pa and 5 Pa magnitudes at frequencies of 0.5 Hz, 1 Hz and 2 Hz for 1 h, 2 h and 4 h of stimulation. Our findings demonstrate that OFF elicits a positive osteogenic response in MSCs in a shear stress magnitude, frequency, and duration dependent manner that is gene specific. Based on the mRNA expression of osteogenic markers Cox2, Runx2 and Opn after short-term fluid flow stimulation, we identified that a regime of 2 Pa shear magnitude and 2 Hz frequency induces the most robust and reliable upregulation in osteogenic gene expression. Furthermore, long-term mechanical stimulation utilising this regime, elicits a significant increase in collagen and mineral deposition when compared to static control demonstrating that mechanical stimuli predicted within the marrow is sufficient to directly drive osteogenesis.     

Flow patterns and wall shear stress distribution in human internal carotid arteries: the geometric effect on the risk for stenoses

It has been widely observed that atherosclerotic stenosis occurs at sites with complex hemodynamics, such as arteries with high curvature or bifurcations. These regions usually have very low or highly oscillatory wall shear stress (WSS). In the present study, 3D sinusoidally pulsatile blood flow through the models of internal carotid artery (ICA) with different geometries was investigated with computational simulation. Three preferred sites of stenoses were found along the carotid siphon with low and highly oscillatory WSS. The risk for stenoses at these sites was scaled with the values of time-averaged WSS and oscillating shear index (OSI). The local risk for stenoses at every preferred site of stenoses was found different between 3 types of ICA, indicating that the geometry of the blood vessel plays significant roles in the atherogenesis. Specifically, the large curvature and planarity of the vessel were found to increase the risk for stenoses, because they tend to lower WSS and elevate OSI. Therefore, the geometric study makes it possible to estimate the stenosis location in the ICA siphon as long as the shape of ICA was measured.     

Differential dependence of stretch and shear stress signaling on caveolin-1 in the vascular wall

The role of caveolae in stretch- versus flow-induced vascular responses was investigated using caveolin 1-deficient [knockout (KO)] mice. Portal veins were stretched longitudinally for 5 min (acute) or 72 h (organ culture). Basal ERK1/2 and Akt phosphorylation were increased in organ-cultured KO veins, as were protein synthesis and vessel wall cross sections. Stretch stimulated acute phosphorylation of ERK1/2 and long-term phosphorylation of focal adhesion kinase (FAK) and cofilin but did not affect Akt phosphorylation. Protein synthesis, and particularly synthesis of smooth muscle differentiation markers, was increased by stretch. These effects did not differ in portal veins from KO and control mice, which also showed the same contractile response to membrane depolarization and inhibition by the Rho kinase inhibitor Y-27632. KO carotid arteries had increased wall cross sections and responded to pressurization (120 mmHg) for 1 h with increased ERK1/2 but not Akt phosphorylation, similar to control arteries. Shear stress by flow for 15 min, on the other hand, increased phosphorylation of Akt in carotids from control but not KO mice. In conclusion, caveolin 1 contributes to low basal ERK1/2 and Akt activity and is required for Akt-dependent signals in response to shear stress (flow) but is not essential for trophic effects of stretch (pressure) in the vascular wall.     

Effect of residual stress and heterogeneity on circumferential stress in the arterial wall

Quantifying the stress distribution through the arterial wall is essential to studies of arterial growth and disease. Previous studies have shown that both residual stress, as measured by opening angle, and differing material properties for the media-intima and the adventitial layers affect the transmural circumferential stress (sigma theta) distribution. Because a lack of comprehensive data on a single species and artery has led to combinations from multiple sources, this study determined the sensitivity of sigma theta to published variations in both opening angle and layer thickness data. We fit material properties to previously published experimental data for pressure-diameter relations and opening angles of rabbit carotid artery, and predicted sigma theta through the arterial wall at physiologic conditions. Using a one-layer model, the ratio of sigma theta at the internal wall to the mean sigma theta decreased from 2.34 to 0.98 as the opening angle increased from 60 to 130 deg. In a two-layer model using a 95 deg opening angle, mean sigma theta in the adventitia increased (112 percent for 25 percent adventitia) and mean sigma theta in the media decreased (47 percent for 25 percent adventitia). These results suggest that both residual stress and wall layers have important effects on transmural stress distribution. Thus, experimental measurements of loading curves, opening angles, and wall composition from the same species and artery are needed to accurately predict the transmural stress distribution in the arterial wall.     

Effects of transmural pressure and wall shear stress on LDL accumulation in the arterial wall: a numerical study using a multilayered model

The accumulation of low-density lipoprotein (LDL) is recognized as one of the main contributors in atherogenesis. Mathematical models have been constructed to simulate mass transport in large arteries and the consequent lipid accumulation in the arterial wall. The objective of this study was to investigate the influences of wall shear stress and transmural pressure on LDL accumulation in the arterial wall by a multilayered, coupled lumen-wall model. The model employs the Navier-Stokes equations and Darcy's Law for fluid dynamics, convection-diffusion-reaction equations for mass balance, and Kedem-Katchalsky equations for interfacial coupling. To determine physiologically realistic model parameters, an optimization approach that searches optimal parameters based on experimental data was developed. Two sets of model parameters corresponding to different transmural pressures were found by the optimization approach using experimental data in the literature. Furthermore, a shear-dependent hydraulic conductivity relation reported previously was adopted. The integrated multilayered model was applied to an axisymmetric stenosis simulating an idealized, mildly stenosed coronary artery. The results show that low wall shear stress leads to focal LDL accumulation by weakening the convective clearance effect of transmural flow, whereas high transmural pressure, associated with hypertension, leads to global elevation of LDL concentration in the arterial wall by facilitating the passage of LDL through wall layers.