In vitro Assessment of Aortic Regurgitation Using Four-Dimensional Flow Magnetic Resonance Imaging.

Aortic regurgitation (AR) refers to backward blood flow from the aorta into the left ventricle (LV) during ventricular diastole. The regurgitant jet arising from the complex shape is characterized by the three-dimensional flow and high-velocity gradient, sometimes limiting an accurate measurement of the regurgitant volume using 2D echocardiography. Recently developed four-dimensional flow magnetic resonance imaging (4D flow MRI) enables three-dimensional volumetric flow measurements, which can be used to accurately quantify the amount of the regurgitation. This study focuses on (i) magnetic resonance compatible AR model fabrication (dilatation, perforation, and prolapse) and (ii) systematic analysis of the performance of 4D flow MRI in AR quantification. The results indicated that the formation of the forward and backward jets over time was highly dependent on the types of AR origin. The amount of regurgitation volume bias for the model types were -7.04%, -33.21%, 6.75%, and 37.04% compared to the ground truth (48 mL) volume measured from the pump stroke volume. The largest error of the regurgitation fraction was around 12%. These results indicate that careful selection of imaging parameters is required when absolute regurgitation volume is important. The suggested in vitro flow phantom can easily be modified to simulate other valvular diseases such as aortic stenosis or bicuspid aortic valve (BAV) and can be used as a standard platform to test different MRI sequences in the future.

In-vitro and In-Vivo Assessment of 4D Flow MRI Reynolds Stress Mapping for Pulsatile Blood Flow.

Imaging hemodynamics play an important role in the diagnosis of abnormal blood flow due to vascular and valvular diseases as well as in monitoring the recovery of normal blood flow after surgical or interventional treatment. Recently, characterization of turbulent blood flow using 4D flow magnetic resonance imaging (MRI) has been demonstrated by utilizing the changes in signal magnitude depending on intravoxel spin distribution. The imaging sequence was extended with a six-directional icosahedral (ICOSA6) flow-encoding to characterize all elements of the Reynolds stress tensor (RST) in turbulent blood flow. In the present study, we aimed to demonstrate the feasibility of full RST analysis using ICOSA6 4D flow MRI under physiological conditions. First, the turbulence analysis was performed through in vitro experiments with a physiological pulsatile flow condition. Second, a total of 12 normal subjects and one patient with severe aortic stenosis were analyzed using the same sequence. The in-vitro study showed that total turbulent kinetic energy (TKE) was less affected by the signal-to-noise ratio (SNR), however, maximum principal turbulence shear stress (MPTSS) and total turbulence production (TP) had a noise-induced bias. Smaller degree of the bias was observed for TP compared to MPTSS. In-vivo study showed that the subject-variability on turbulence quantification was relatively low for the consistent scan protocol. The in vivo demonstration of the stenosis patient showed that the turbulence analysis could clearly distinguish the difference in all turbulence parameters as they were at least an order of magnitude larger than those from the normal subjects.

Endothelial cell monolayer-based microfluidic systems mimicking complex in vivo microenvironments for the study of leukocyte dynamics in inflamed blood vessels.

In response to inflammatory signals, leukocytes circulating in blood vessels undergo dynamic interaction with endothelium lining blood vessel walls, known as leukocyte adhesion cascade, to leave the blood vessels, and infiltrate the inflamed tissue. During leukocyte extravasation, leukocytes recognize and respond to various biophysical and biochemical cues present in the complex microenvironments of inflamed blood vessels to find optimal pathways. Although advances in intravital imaging of live animals have enabled us to observe leukocyte dynamics during extravasation, in vitro model systems mimicking complex in vivo microenvironments are still needed for mechanistic studies. A parallel-plate flow chamber assembled by placing a fluidic chamber on an endothelial cell (EC) monolayer has been widely used as an in vitro model to study leukocyte dynamics in inflamed blood vessels. Although this is a simple yet powerful model providing well-defined flow conditions, a parallel-plate flow chamber lacks the complex microenvironments of inflamed blood vessels. In this article, we first describe the basic design, assembly, and operation principles of a parallel-plate flow chamber. Then, we present methods of incorporating various features of in vivo microenvironments into parallel-plate flow chambers, including EC alignment using nanogrooved surfaces, insertion of a stenotic structure for complex flow generation, and extension to 3D blood vessel/inflamed tissue models.

Effect of pannus formation on the prosthetic heart valve: In vitro demonstration using particle image velocimetry.

Although hemodynamic influence of the subprosthetic tissue, termed as pannus, may contribute to prosthetic aortic valve dysfunction, the relationship between pannus extent and hemodynamics in the prosthetic valve has rarely been reported. We investigated the fluid dynamics of pannus formation using in vitro experiments with particle image velocimetry. Subvalvular pannus formation caused substantial changes in prosthetic valve transvalvular peak velocity, transvalvular pressure gradient (TPG) and opening angle. Maximum flow velocity and corresponding TPG were mostly affected by pannus width. When the pannus width was 25% of the valve diameter, pannus formation elevated TPG to >2.5 times higher than that without pannus formation. Opening dysfunction was observed only for a pannus involvement angle of 360°. Although circumferential pannus with an involvement angle of 360° decreased the opening angle of the valve from approximately 82° to 58°, eccentric pannus with an involvement angle of 180° did not induce valve opening dysfunction. The pannus involvement angle largely influenced the velocity flow field at the aortic sinus and corresponding hemodynamic indices, including wall shear stress, principal shear stress and viscous energy loss distributions. Substantial discrepancy between the velocity-based TPG estimation and direct pressure measurements was observed for prosthetic valve flow with pannus formation.

Hemodynamic characteristics of flow around a deformable stenosis.

Clinical studies reported that some vulnerable stenoses deformed their shape in a blood vessel based on flow condition. However, the effects of shape variation on flow characteristics remain unclear. The flow characteristics are known to affect vulnerable stenosis rupture and fractional flow reserve (FFR) value which has been widely used as a diagnostic tool for stenosis. Vulnerable stenosis rupture occurs when the structural stress exerted on a fibrous cap exceeds its tolerable threshold. The stress magnitude is determined from the spatial distribution of static pressure around the stenosis. In the present study, the static pressure distribution and the FFR value in deformable stenosis were investigated with related other flow characteristics. Two phantom models were fabricated to mimic deformable and nondeformable stenoses using polydimethylsiloxane. The flow characteristics were observed under a steady-flow condition at three Reynolds numbers (Re=500, 1000, 1500) using a particle image velocimetry. The pressure drop across the stenosis models were measured using a pressure sensor to determine effects of shape deformation on FFR value. Shape variations and jet deflections were clearly observed in the deformable stenosis model, and the effective severity of the stenosis increased up to 17.2%. The shape variations of deformable stenosis model increased the static pressure difference at the upstream and downstream sides of the stenosis. The pressure drop across the deformable stenosis model was significantly higher than that of the nondeformable stenosis model. The present results substantiate that stenosis deformability should be carefully considered to diagnose the rupture of vulnerable stenosis.

Homocysteine-induced peripheral microcirculation dysfunction in zebrafish and its attenuation by L-arginine.

Elevated blood homocysteine (Hcy) level is frequently observed in aged individuals and those with age-related vascular diseases. However, its effect on peripheral microcirculation is still not fully understood. Using in vivo zebrafish model, the degree of Hcy-induced peripheral microcirculation dysfunction is assessed in this study with a proposed dimensionless velocity parameter [Formula: see text], where [Formula: see text] and [Formula: see text] represent the peripheral microcirculation perfusion and the systemic perfusion levels, respectively. The ratio of the peripheral microcirculation perfusion to the systemic perfusion is largely decreased due to peripheral accumulation of neutrophils, while the systemic perfusion is relatively preserved by increased blood supply from subintestinal vein. Pretreatment with L-arginine attenuates the effects of Hcy on peripheral microcirculation and reduces the peripheral accumulation of neutrophils. Given its convenience, high reproducibility of the observation site, non-invasiveness, and the ease of drug treatment, the present zebrafish model with the proposed parameters will be used as a useful drug screening platform for investigating the pathophysiology of Hcy-induced microvascular diseases.

Flow characteristics around proximal and distal stenoses in a series of tandem stenosed vessels.

The flow characteristics around the proximal and distal stenoses in tandem vessel models are experimentally investigated with varying flow rates (Q=0.25, 0.5, 1.0L/min), interspacing distances (L=3, 6, 10 of diameter D) and severities (S=50%, 75% reduction in diameter). When the interspacing L is larger than 10 D, no fluid-dynamic interaction is observed. The flow between the proximal and distal stenoses becomes stabilized (turbulence intensity of <3%) as the interspacing distance decreases. When the severity S is 75%, the transition from laminar to turbulent flow occurs at a flow rate higher than 0.5L/min, although the interspacing distance L is 3 D. Formation of recirculation flow is restricted by the presence of distal stenosis as the interspacing distance decreases. In this case, the flow between the stenoses is focused on the central region. The center-line velocity at the neck of the distal stenosis is approximately 10-15% higher than that of the proximal stenosis with equal severity of S=50%. When the inlet flow is center-focused, the lengths of the recirculation and the jet core behind the distal stenosis increase with decrease in interspacing distance L. When the inlet flow is turbulent, the transition from laminar to turbulent flow occurs early as the interspacing distance L is reduced. When the upstream proximal stenosis exhibits increased severity, the pressure drop is measured to be 20% compared with that when the severity of the downstream distal stenosis is increased at the flow rate of Q=1.0L/min.

Post-stenotic Recirculating Flow May Cause Hemodynamic Perforator Infarction.

BACKGROUND AND PURPOSE: The primary mechanism underlying paramedian pontine infarction (PPI) is atheroma obliterating the perforators. Here, we encountered a patient with PPI in the post-stenotic area of basilar artery (BA) without a plaque, shown by high-resolution magnetic resonance imaging (HR-MRI). We performed an experiment using a 3D-printed BA model and a particle image velocimetry (PIV) to explore the hemodynamic property of the post-stenotic area and the mechanism of PPI. METHODS: 3D-model of a BA stenosis was reconstructed with silicone compound using a 3D-printer based on the source image of HR-MRI. Working fluid seeded with fluorescence particles was used and the velocity of those particles was measured horizontally and vertically. Furthermore, microtubules were inserted into the posterior aspect of the model to measure the flow rates of perforators (pre-and post-stenotic areas). The flow rates were compared between the microtubules. RESULTS: A recirculating flow was observed from the post-stenotic area in both directions forming a spiral shape. The velocity of the flow in these regions of recirculation was about one-tenth that of the flow in other regions. The location of recirculating flow well corresponded with the area with low-signal intensity at the time-of-flight magnetic resonance angiography and the location of PPI. Finally, the flow rate through the microtubule inserted into the post-stenotic area was significantly decreased comparing to others (P<0.001). CONCLUSIONS: Perforator infarction may be caused by a hemodynamic mechanism altered by stenosis that induces a recirculation flow. 3D-printed modeling and PIV are helpful understanding the hemodynamics of intracranial stenosis.

Turbulent Kinetic Energy Measurement Using Phase Contrast MRI for Estimating the Post-Stenotic Pressure Drop: In Vitro Validation and Clinical Application.

BACKGROUND: Although the measurement of turbulence kinetic energy (TKE) by using magnetic resonance imaging (MRI) has been introduced as an alternative index for quantifying energy loss through the cardiac valve, experimental verification and clinical application of this parameter are still required. OBJECTIVES: The goal of this study is to verify MRI measurements of TKE by using a phantom stenosis with particle image velocimetry (PIV) as the reference standard. In addition, the feasibility of measuring TKE with MRI is explored. METHODS: MRI measurements of TKE through a phantom stenosis was performed by using clinical 3T MRI scanner. The MRI measurements were verified experimentally by using PIV as the reference standard. In vivo application of MRI-driven TKE was explored in seven patients with aortic valve disease and one healthy volunteer. Transvalvular gradients measured by MRI and echocardiography were compared. RESULTS: MRI and PIV measurements of TKE are consistent for turbulent flow (0.666 < R2 < 0.738) with a mean difference of -11.13 J/m3 (SD = 4.34 J/m3). Results of MRI and PIV measurements differ by 2.76 ± 0.82 cm/s (velocity) and -11.13 ± 4.34 J/m3 (TKE) for turbulent flow (Re > 400). The turbulence pressure drop correlates strongly with total TKE (R2 = 0.986). However, in vivo measurements of TKE are not consistent with the transvalvular pressure gradient estimated by echocardiography. CONCLUSIONS: These results suggest that TKE measurement via MRI may provide a potential benefit as an energy-loss index to characterize blood flow through the aortic valve. However, further clinical studies are necessary to reach definitive conclusions regarding this technique.

Structural design of a double-layered porous hydrogel for effective mass transport.

Mass transport in porous materials is universal in nature, and its worth attracts great attention in many engineering applications. Plant leaves, which work as natural hydraulic pumps for water uptake, have evolved to have the morphological structure for fast water transport to compensate large water loss by leaf transpiration. In this study, we tried to deduce the advantageous structural features of plant leaves for practical applications. Inspired by the tissue organization of the hydraulic pathways in plant leaves, analogous double-layered porous models were fabricated using agarose hydrogel. Solute transport through the hydrogel models with different thickness ratios of the two layers was experimentally observed. In addition, numerical simulation and theoretical analysis were carried out with varying porosity and thickness ratio to investigate the effect of structural factors on mass transport ability. A simple parametric study was also conducted to examine unveiled relations between structural factors. As a result, the porosity and thickness ratio of the two layers are found to govern the mass transport ability in double-layered porous materials. The hydrogel models with widely dispersed pores at a fixed porosity, i.e., close to a homogeneously porous structure, are mostly turned out to exhibit fast mass transport. The present results would provide a new framework for fundamental design of various porous structures for effective mass transport.