Ultrasound Shear Wave Attenuation Imaging for Grading Liver Steatosis in Volunteers and Patients With Non-alcoholic Fatty Liver Disease: A Pilot Study.

OBJECTIVE: The aims of the work described here were to assess shear wave attenuation (SWA) in volunteers and patients with non-alcoholic fatty liver disease (NAFLD) and compare its diagnostic performance with that of shear wave dispersion (SWD), magnetic resonance imaging (MRI) proton density fat fraction (PDFF) and biopsy. METHODS: Forty-nine participants (13 volunteers and 36 NAFLD patients) were enrolled. Ultrasound and MRI examinations were performed in all participants. Biopsy was also performed in patients. SWA was used to assess histopathology grades as potential confounders. The areas under curves (AUCs) of SWA, SWD and MRI-PDFF were assessed in different steatosis grades by biopsy. Youden's thresholds of SWA were obtained for steatosis grading while using biopsy or MRI-PDFF as the reference standard. RESULTS: Spearman's correlations of SWA with histopathology (steatosis, inflammation, ballooning and fibrosis) were 0.89, 0.73, 0.62 and 0.31, respectively. Multiple linear regressions of SWA confirmed the correlation with steatosis grades (adjusted R2 = 0.77, p < 0.001). The AUCs of MRI-PDFF, SWA and SWD were respectively 0.97, 0.99 and 0.94 for S0 versus ≥S1 (p > 0.05); 0.94, 0.98 and 0.78 for ≤S1 versus ≥S2 (both MRI-PDFF and SWA were higher than SWD, p < 0.05); and 0.90, 0.93 and 0.68 for ≤S2 versus S3 (both SWA and MRI-PDFF were higher than SWD, p < 0.05). SWA's Youden thresholds (Np/m/Hz) (sensitivity, specificity) for S0 versus ≥S1, ≤S1 versus ≥S2 and ≤S2 versus S3 were 1.05 (1.00, 0.92), 1.37 (0.96, 0.96) and 1.51 (0.83, 0.87), respectively. These values were 1.16 (1.00, 0.81), 1.49 (0.91, 0.82) and 1.67 (0.87, 0.92) when considering MRI-PDFF as the reference standard. CONCLUSION: In this pilot study, SWA increased with increasing steatosis grades, and its diagnostic performance was higher than that of SWD but equivalent to that of MRI-PDFF.

Ultrafast Myocardial Principal Strain Ultrasound Elastography During Stress Tests: In Vitro Validation and In Vivo Feasibility.

Objective myocardial contractility assessment during stress tests aims to improve the diagnosis of myocardial ischemia. Tissue Doppler imaging (TDI) or optical flow (OF) speckle tracking echocardiography (STE) has been used to quantify myocardial contractility at rest. However, this is more challenging during stress tests due to image decorrelation at high heart rates. Moreover, stress tests imply a high frame rate which leads to a limited lateral field of view. Therefore, a large lateral field-of-view robust ultrafast myocardial regularized OF-TDI principal strain estimator has been developed for high-frame-rate echocardiography of coherently compounded transmitted diverging waves. The feasibility and accuracy of the proposed estimator were validated in vitro (using sonomicrometry as the gold standard) and in vivo stress experiments. Compared with OF strain imaging, the proposed estimator improved the accuracy of principal major and minor strains during stress tests, with an average contrast-to-noise ratio improvement of 4.4 ± 2.7 dB ( p -value < 0.01). Moreover, there was a significant correlation and a very close agreement between the proposed estimator and sonomicrometry for tested heart rates between 60 and 180 beats per minute (bpm). The averages ± standard deviations (STD) of R2 and biases ± STD between them were 0.96 ± 0.04 ( p -value < 0.01) and 0.01 ± 0.03% in the axial direction, respectively; and 0.94 ± 0.02 ( p -value < 0.01) and 0.04 ± 0.06% in the lateral direction, respectively. These results suggest that the proposed estimator could be useful clinically to provide an accurate and quantitative 2-D large lateral field-of-view myocardial strain assessment at high heart rates during stress echocardiography.

Quantitative ultrasound, elastography, and machine learning for assessment of steatosis, inflammation, and fibrosis in chronic liver disease.

OBJECTIVE: To develop a quantitative ultrasound (QUS)- and elastography-based model to improve classification of steatosis grade, inflammation grade, and fibrosis stage in patients with chronic liver disease in comparison with shear wave elastography alone, using histopathology as the reference standard. METHODS: This ancillary study to a prospective institutional review-board approved study included 82 patients with non-alcoholic fatty liver disease, chronic hepatitis B or C virus, or autoimmune hepatitis. Elastography measurements, homodyned K-distribution parametric maps, and total attenuation coefficient slope were recorded. Random forests classification and bootstrapping were used to identify combinations of parameters that provided the highest diagnostic accuracy. Receiver operating characteristic (ROC) curves were computed. RESULTS: For classification of steatosis grade S0 vs. S1-3, S0-1 vs. S2-3, S0-2 vs. S3, area under the receiver operating characteristic curve (AUC) were respectively 0.60, 0.63, and 0.62 with elasticity alone, and 0.90, 0.81, and 0.78 with the best tested model combining QUS and elastography features. For classification of inflammation grade A0 vs. A1-3, A0-1 vs. A2-3, A0-2 vs. A3, AUCs were respectively 0.56, 0.62, and 0.64 with elasticity alone, and 0.75, 0.68, and 0.69 with the best model. For classification of liver fibrosis stage F0 vs. F1-4, F0-1 vs. F2-4, F0-2 vs. F3-4, F0-3 vs. F4, AUCs were respectively 0.66, 0.77, 0.72, and 0.74 with elasticity alone, and 0.72, 0.77, 0.77, and 0.75 with the best model. CONCLUSION: Random forest models incorporating QUS and shear wave elastography increased the classification accuracy of liver steatosis, inflammation, and fibrosis when compared to shear wave elastography alone.

Deformability of ascending thoracic aorta aneurysms assessed using ultrafast ultrasound and a principal strain estimator: In vitro evaluation and in vivo feasibility.

BACKGROUND: Noninvasive vascular strain imaging under conventional line-by-line scanning has a low frame rate and lateral resolution and depends on the coordinate system. It is thus affected by high deformations due to image decorrelation between frames. PURPOSE: To develop an ultrafast time-ensemble regularized tissue-Doppler optical-flow principal strain estimator for aorta deformability assessment in a long-axis view. METHODS: This approach alleviated the impact of lateral resolution using image compounding and that of the coordinate system dependency using principal strain. Accuracy and feasibility were evaluated in two aorta-mimicking phantoms first, and then in four age-matched individuals with either a normal aorta or a pathological ascending thoracic aorta aneurysm (TAA). RESULTS: Instantaneous aortic maximum and minimum principal strain maps and regional accumulated strains during each cardiac cycle were estimated at systolic and diastolic phases to characterize the normal aorta and TAA. In vitro, principal strain results matched sonomicrometry measurements. In vivo, a significant decrease in maximum and minimum principal strains was observed in TAA cases, whose range was respectively 7.9 ± 6.4% and 8.2 ± 2.6% smaller than in normal aortas. CONCLUSIONS: The proposed principal strain estimator showed an ability to potentially assess TAA deformability, which may provide an individualized and reliable evaluation method for TAA rupture risk assessment.

Shear Wave Elastography and Quantitative Ultrasound as Biomarkers to Characterize Deep Vein Thrombosis In Vivo.

OBJECTIVE: Investigate shear wave elastography (SWE) and quantitative ultrasound (QUS) parameters in patients hospitalized for lower limb deep vein thrombosis (DVT). METHOD: Sixteen patients with DVT were recruited and underwent SWE and radiofrequency data acquisitions for QUS on day 0, day 7, and day 30 after the beginning of symptoms, in both proximal and distal zones of the clot identified on B-mode scan. SWE and QUS features were computed to differentiate between thrombi at day 0, day 7, and day 30 following treatment with heparin or oral anticoagulant. The Young's modulus from SWE was computed, as well as QUS homodyned K-distribution (HKD) parameters reflecting blood clot structure. Median and interquartile range of SWE and QUS parameters within clot were taken as features. RESULTS: In the proximal zone of the clot, the HKD ratio of coherent-to-diffuse backscatter median showed a significant decrease from day 7 to day 30 (P = .036), while the HKD ratio of diffuse-to-total backscatter median presented a significant increase from day 7 to day 30 (P = .0491). In the distal zone of the clot, the HKD normalized intensity of the echo envelope median showed a significant increase from day 0 to day 30 (P = .0062). No SWE features showed statistically significant differences over time. Nonetheless, a trend of lower median of Young's modulus within clot for patients who developed a pulmonary embolism was observed. CONCLUSION: QUS features may be relevant to characterize clot's evolution over time. Further analysis of their clinical interpretation and validation on a larger dataset would deserve to be studied.

Impact of Applying a Skin Compression With the Ultrasound Probe on Carotid Artery Strain Elastography.

OBJECTIVE: To study the impact of varying the external compression exerted by the ultrasound probe when performing a carotid strain elastography exam. METHODS: Nine healthy volunteers (mean age 43 years ±13 years; 6 men) underwent a vascular ultrasound elastography exam using a custom made sound feedback handle embedding the probe, and allowing the sonographer to adjust the applied compression. A clinical standard practice (SP) force was first recorded, and then predetermined compression (PDC) forces were applied, ranging from 0 to 5 N for the left common carotid artery (CCA) or 2-12 N for the left internal carotid artery (ICA). Six carotid elastography features, namely maximum and cumulated axial strains, maximum and cumulated shear strains, cumulated axial translation, and cumulated lateral translation were assessed with noninvasive vascular elastography (NIVE) on near and far walls of carotids. The carotid intima media thickness (IMT) and diameter were also measured. RESULTS: All elastography features on the near wall of both CCA and ICA decreased statistically significantly as the PDC force increased; this association was also observed for half of the features on the far wall. Three NIVE features at the lowest PDC force (out of 72 that were tested) were statistically significantly different than values at the SP force. Overall, NIVE showed some variance to probe compression with linear regression slopes revealing changes of 10.1%-45.6% in magnitude over the whole compression range on both walls. The maximum IMT for the ICA near wall, and carotid lumen diameters of both CCA and ICA were statistically significantly associated with PDC forces; these features underwent a decrease of 10.2%, 36.2%, and 17.6%, respectively, over the whole range of PDC force increase. Other IMT measurements were not statistically significantly associated with applied PDC forces. CONCLUSION: These results suggest the need of technical guidelines for carotid strain elastography. Using the lowest probe compression while allowing a good B-mode image quality is recommended to improve the robustness of NIVE measurements.

Parameterized Strain Estimation for Vascular Ultrasound Elastography With Sparse Representation.

Ultrasound vascular strain imaging has shown its potential to interrogate the motion of the vessel wall induced by the cardiac pulsation for predicting plaque instability. In this study, a sparse model strain estimator (SMSE) is proposed to reconstruct a dense strain field at a high resolution, with no spatial derivatives, and a high computation efficiency. This sparse model utilizes the highly-compacted property of discrete cosine transform (DCT) coefficients, thereby allowing to parameterize displacement and strain fields with truncated DCT coefficients. The derivation of affine strain components (axial and lateral strains and shears) was reformulated into solving truncated DCT coefficients and then reconstructed with them. Moreover, an analytical solution was derived to reduce estimation time. With simulations, the SMSE reduced estimation errors by up to 50% compared with the state-of-the-art window-based Lagrangian speckle model estimator (LSME). The SMSE was also proven to be more robust than the LSME against global and local noise. For in vitro and in vivo tests, residual strains assessing cumulated errors with the SMSE were 2 to 3 times lower than with the LSME. Regarding computation efficiency, the processing time of the SMSE was reduced by 4 to 25 times compared with the LSME, according to simulations, in vitro and in vivo results. Finally, phantom studies demonstrated the enhanced spatial resolution of the proposed SMSE algorithm against LSME.

Anthropomorphic and biomechanical mockup for abdominal aortic aneurysm.

Abdominal aortic aneurysm (AAA) is an asymptomatic condition due to the dilation of abdominal aorta along with progressive wall degeneration, where rupture of AAA is life-threatening. Failures of AAA endovascular repair (EVAR) reflect our inadequate knowledge about the complex interaction between the aortic wall and medical devices. In this regard, we are presenting a hydrogel-based anthropomorphic mockup (AMM) to better understand the biomechanical constraints during EVAR. By adjusting the cryogenic treatments, we tailored the hydrogel to mimic the mechanical behavior of human AAA wall, thrombus and abdominal fat. A specific molding sequence and a pressurizing system were designed to reproduce the geometrical and diseased characteristics of AAA. A mechanically, anatomically and pathologically realistic AMM for AAA was developed for the first time, EVAR experiments were then performed with and without the surrounding fat. Substantial displacements of the aortic centerlines and vessel expansion were observed in the case without surrounding fat, revealing an essential framework created by the surrounding fat to account for the interactions with medical devices. In conclusion, the importance to consider surrounding tissue for the global deformation of AAA during EVAR was highlighted. Furthermore, potential use of this AMM for medical training was also suggested.

Pilot clinical study of quantitative ultrasound spectroscopy measurements of erythrocyte aggregation within superficial veins.

BACKGROUND: An enhanced inflammatory response is a trigger to the production of blood macromolecules involved in abnormally high levels of erythrocyte aggregation. OBJECTIVE: This study aimed at demonstrating for the first time the clinical feasibility of a non-invasive ultrasound-based erythrocyte aggregation quantitative measurement method for potential application in critical care medicine. METHODS: Erythrocyte aggregation was evaluated using modeling of the backscatter coefficient with the Structure Factor Size and Attenuation Estimator (SFSAE). SFSAE spectral parameters W (packing factor) and D (mean aggregate diameter) were measured within the antebrachial vein of the forearm and tibial vein of the leg in 50 healthy participants at natural flow and reduced flow controlled by a pressurized bracelet. Blood samples were also collected to measure erythrocyte aggregation ex vivo with an erythroaggregometer (parameter S10). RESULTS: W and Din vivo measurements were positively correlated with the ex vivoS10 index for both measurement sites and shear rates (correlations between 0.35-0.81, p < 0.05). Measurement at low shear rate was found to increase the sensitivity and reliability of this non-invasive measurement method. CONCLUSIONS: We behold that the SFSAE method presents systemic measures of the erythrocyte aggregation level, since results on upper and lower limbs were highly correlated.

Added Value of Quantitative Ultrasound and Machine Learning in BI-RADS 4-5 Assessment of Solid Breast Lesions.

The purpose of this study was to evaluate various combinations of 13 features based on shear wave elasticity (SWE), statistical and spectral backscatter properties of tissues, along with the Breast Imaging Reporting and Data System (BI-RADS), for classification of solid breast lesions at ultrasonography by means of random forests. One hundred and three women with 103 suspicious solid breast lesions (BI-RADS categories 4-5) were enrolled. Before biopsy, additional SWE images and a cine sequence of ultrasound images were obtained. The contours of lesions were delineated, and parametric maps of the homodyned-K distribution were computed on three regions: intra-tumoral, supra-tumoral and infra-tumoral zones. Maximum elasticity and total attenuation coefficient were also extracted. Random forests yielded receiver operating characteristic (ROC) curves for various combinations of features. Adding BI-RADS category improved the classification performance of other features. The best result was an area under the ROC curve of 0.97, with 75.9% specificity at 98% sensitivity.

Reconstruction of Viscosity Maps in Ultrasound Shear Wave Elastography.

Change in viscoelastic properties of biological tissues may often be symptomatic of a dysfunction that can be correlated to tissue pathology. Shear wave elastography is an imaging method mainly used to assess stiffness but with the potential to measure viscoelasticity of biological tissues. This can enable tissue characterization; and thus, can be used as a marker to improve diagnosis of pathological lesions. In this study, a frequency-shift method based framework is presented for the reconstruction of viscosity by analyzing the spectral properties of acoustic radiation force-induced shear waves. The aim of the study was to investigate the feasibility of viscosity reconstruction maps in homogeneous as well as heterogeneous samples. Experiments were performed in four in vitro phantoms, two ex vivo porcine liver samples, two ex vivo fatty duck liver samples, and one in vivo fatty goose liver. Successful viscosity maps were reconstructed in homogeneous and heterogeneous phantoms with embedded mechanical inclusions having different geometries. Quantitative values of viscosity obtained for two porcine liver tissues, two fatty duck liver samples, and one goose fatty liver were (mean ± SD) 0.61 ± 0.21, 0.52 ± 0.35; 1.28 ± 0.54, 1.36 ± 0.73, and 1.67 ± 0.70 Pa.s, respectively.

Atherosclerotic carotid bifurcation phantoms with stenotic soft inclusions for ultrasound flow and vessel wall elastography imaging.

As the complexity of ultrasound signal processing algorithms increases, it becomes more difficult to demonstrate their added value and thus robust validation strategies are required. We propose a method of manufacturing ultrasonic vascular phantoms mimicking an atheromatous plaque in an internal carotid artery bifurcation for applications in flow imaging and elastography. During the fabrication process, a soft inclusion mimicking a stenotic lipid pool was embedded within the vascular wall. Mechanical testing measured Young's moduli of the vascular wall and soft inclusion at 342 ± 25 kPa and 17 ± 3 kPa, respectively. B-mode, color Doppler, power Doppler, shear wave elastography, and strain elastography images of the different phantoms were produced to show the validity of the fabrication process. Because of their realistic geometries and mechanical properties, those phantoms may become advantageous for fluid-structure experimental modeling and validation of new ultrasound-based imaging technologies.

Investigation of out-of-plane motion artifacts in 2D noninvasive vascular ultrasound elastography.

Ultrasound noninvasive vascular elastography (NIVE) has shown its potential to measure strains of carotid arteries to predict plaque instability. When two-dimensional (2D) strain estimation is performed, either in longitudinal or cross-sectional view, only in-plane motions are considered. The motions in elevation direction (i.e. perpendicular to the imaging plane), can induce estimation artifacts affecting the accuracy of 2D NIVE. The influence of such out-of-plane motions on the performance of axial strain and axial shear strain estimations has been evaluated in this study. For this purpose, we designed a diseased carotid bifurcation phantom with a 70% stenosis and an in vitro experimental setup to simulate orthogonal out-of-plane motions of 1 mm, 2 mm and 3 mm. The Lagrangian speckle model estimator (LSME) was used to estimate axial strains and shears under pulsatile conditions. As anticipated, in vitro results showed more strain estimation artifacts with increasing magnitudes of motions in elevation. However, even with an out-of-plane motion of 2.0 mm, strain and shear estimations having inter-frame correlation coefficients higher than 0.85 were obtained. To verify findings of in vitro experiments, a clinical LSME dataset obtained from 18 participants with carotid artery stenosis was used. Deduced out-of-plane motions (ranging from 0.25 mm to 1.04 mm) of the clinical dataset were classified into three groups: small, moderate and large elevational motions. Clinical results showed that pulsatile time-varying strains and shears remained reproducible for all motion categories since inter-frame correlation coefficients were higher than 0.70, and normalized cross-correlations (NCC) between radiofrequency (RF) images were above 0.93. In summary, the performance of LSME axial strain and shear estimations appeared robust in the presence of out-of-plane motions (<2 mm) as encountered during clinical ultrasound imaging.

Estimation of polydispersity in aggregating red blood cells by quantitative ultrasound backscatter analysis.

Quantitative ultrasound techniques based on the backscatter coefficient (BSC) have been commonly used to characterize red blood cell (RBC) aggregation. Specifically, a scattering model is fitted to measured BSC and estimated parameters can provide a meaningful description of the RBC aggregates' structure (i.e., aggregate size and compactness). In most cases, scattering models assumed monodisperse RBC aggregates. This study proposes the Effective Medium Theory combined with the polydisperse Structure Factor Model (EMTSFM) to incorporate the polydispersity of aggregate size. From the measured BSC, this model allows estimating three structural parameters: the mean radius of the aggregate size distribution, the width of the distribution, and the compactness of the aggregates. Two successive experiments were conducted: a first experiment on blood sheared in a Couette flow device coupled with an ultrasonic probe, and a second experiment, on the same blood sample, sheared in a plane-plane rheometer coupled to a light microscope. Results demonstrated that the polydisperse EMTSFM provided the best fit to the BSC data when compared to the classical monodisperse models for the higher levels of aggregation at hematocrits between 10% and 40%. Fitting the polydisperse model yielded aggregate size distributions that were consistent with direct light microscope observations at low hematocrits.

Accuracy of speckle tracking in the context of stress echocardiography in short axis view: An in vitro validation study.

AIM: This study aimed to test the accuracy of a speckle tracking algorithm to assess myocardial deformation in a large range of heart rates and strain magnitudes compared to sonomicrometry. METHODS AND RESULTS: Using a tissue-mimicking phantom with cyclic radial deformation, radial strain derived from speckle tracking (RS-SpT) of the upper segment was assessed in short axis view by conventional echocardiography (Vivid q, GE) and post-processed with clinical software (EchoPAC, GE). RS-SpT was compared with radial strain measured simultaneously by sonomicrometers (RS-SN). Radial strain was assessed with increasing deformation rates (60 to 160 beats/min) and increasing pulsed volumes (50 to 100 ml/beat) to simulate physiological changes occurring during stress echocardiography. There was a significant correlation (R2 = 0.978, P <0.001) and a close agreement (bias ± 2SD, 0.39 ± 1.5%) between RS-SpT and RS-SN. For low strain values (<15%), speckle tracking showed a small but significant overestimation of radial strain compared to sonomicrometers. Two-way analysis of variance did not show any significant effect of the deformation rate. For RS-SpT, the feasibility was excellent and the intra- and inter-observer variability were low (the intraclass correlation coefficients were 0.96 and 0.97, respectively). CONCLUSIONS: Speckle tracking demonstrated a good correlation with sonomicrometry for the assessment of radial strain independently of the heart rate and strain magnitude in a physiological range of values. Though speckle tracking seems to be a reliable and reproducible technique to assess myocardial deformation variations during stress echocardiography, further studies are mandated to analyze the impact of angulated and artefactual out-of-plane motions and inter-vendor variability.

Acoustic radiation force induced resonance elastography of coagulating blood: theoretical viscoelasticity modeling and ex-vivo experimentation.

Deep vein thrombosis is a common vascular disease that can lead to pulmonary embolism and death. The early diagnosis and clot age staging are important parameters for reliable therapy planning. This article presents an acoustic radiation force induced resonance elastography method for the viscoelastic characterization of clotting blood. The physical concept of this method relies on the mechanical resonance of the blood clot occurring at specific frequencies. Resonances are induced by focusing ultrasound beams inside the sample under investigation. Coupled to an analytical model of wave scattering, the ability of the proposed method to characterize the viscoelasticity of a mimicked venous thrombosis in the acute phase is demonstrated. Experiments with a gelatin-agar inclusion sample of known viscoelasticity are performed for validation and establishment of the proof of concept. In addition, an inversion method is applied in-vitro for the kinetic monitoring of the blood coagulation process of six human blood samples obtained from two volunteers. The computed elasticity and viscosity values of blood samples at the end of the 90 min kinetics were estimated at 411 ± 71 Pa and 0.25 ± 0.03 Pa.s for volunteer #1, and 387 ± 35 Pa and 0.23 ± 0.02 Pa.s for volunteer #2, respectively. The proposed method allowed reproducible time-varying thrombus viscoelastic measurements from samples having physiological dimensions.

Noninvasive Vascular Modulography Method for Imaging the Local Elasticity of Atherosclerotic Plaques: Simulation and In Vitro Vessel Phantom Study.

Mechanical and morphological characterization of atherosclerotic lesions in carotid arteries remains an essential step for the evaluation of rupture prone plaques and the prevention of strokes. In this paper, we propose a noninvasive vascular imaging modulography (NIV-iMod) method, which is capable of reconstructing a heterogeneous Young's modulus distribution of a carotid plaque from the Von Mises strain elastogram. Elastograms were computed with noninvasive ultrasound images using the Lagrangian speckle model estimator and a dynamic segmentation-optimization procedure to highlight mechanical heterogeneities. This methodology, based on continuum mechanics, was validated in silico with finite-element model strain fields and ultrasound simulations, and in vitro with polyvinyl alcohol cryogel phantoms based on magnetic resonance imaging geometries of carotid plaques. In silico, our results show that the NiV-iMod method: 1) successfully detected and quantified necrotic core inclusions with high positive predictive value (PPV) and sensitivity value (SV) of 81±10% and 91±6%; 2) quantified Young's moduli of necrotic cores, fibrous tissues, and calcium inclusions with mean values of 32±23, 515±30, and 3160±218 kPa (ground true values are 10, 600, and 5000 kPa); and 3) overestimated the cap thickness by . In vitro, the PPV and SV for detecting soft inclusions were 60±21% and 88±9%, and Young's modulus mean values of mimicking lipid, fibrosis, and calcium were 34±19, 193±14, and 649±118 kPa (ground true values are 25±3, 182±21, and 757±87 kPa).

Protocol for Robust In Vivo Measurements of Erythrocyte Aggregation Using Ultrasound Spectroscopy.

Erythrocyte aggregation is a non-specific marker of acute and chronic inflammation. Although it is usual to evaluate this phenomenon from blood samples analyzed in laboratory instruments, in vivo real-time assessment of aggregation is possible with spectral ultrasound techniques. However, variable blood flow can affect the interpretation of acoustic measures. Therefore, flow standardization is required. Two techniques of flow standardization were evaluated with porcine and equine blood samples in Couette flow. These techniques consisted in either stopping the flow or reducing it. Then, the sensibility and repeatability of the retained method were evaluated in 11 human volunteers. We observed that stopping the flow compromised interpretation and repeatability. Conversely, maintaining a low flow provided repeatable measures and could distinguish between normal and high extents of erythrocyte aggregation. Agreement was observed between in vivo and ex vivo measures of the phenomenon (R2 = 82.7%, p value < 0.0001). These results support the feasibility of assessing in vivo erythrocyte aggregation in humans by quantitative ultrasound means.

Influence of erythrocyte aggregation on radial migration of platelet-sized spherical particles in shear flow.

Blood platelets when activated are involved in the mechanisms of hemostasis and thrombosis, and their migration toward injured vascular endothelium necessitates interaction with red blood cells (RBCs). Rheology co-factors such as a high hematocrit and a high shear rate are known to promote platelet mass transport toward the vessel wall. Hemodynamic conditions promoting RBC aggregation may also favor platelet migration, particularly in the venous system at low shear rates. The aim of this study was to confirm experimentally the impact of RBC aggregation on platelet-sized micro particle migration in a Couette flow apparatus. Biotin coated micro particles were mixed with saline or blood with different aggregation tendencies, at two shear rates of 2 and 10s-1 and three hematocrits ranging from 20 to 60%. Streptavidin membranes were respectively positioned on the Couette static and rotating cylinders upon which the number of adhered fluorescent particles was quantified. The platelet-sized particle adhesion on both walls was progressively enhanced by increasing the hematocrit (p<0.001), reducing the shear rate (p<0.001), and rising the aggregation of RBCs (p<0.001). Particle count was minimum on the stationary cylinder when suspended in saline at 2s-1 (57±33), and maximum on the rotating cylinder at 60% hematocrit, 2s-1 and the maximum dextran-induced RBC aggregation (2840±152). This fundamental study is confirming recent hypotheses on the role of RBC aggregation on venous thrombosis, and may guide molecular imaging protocols requiring injecting active labeled micro particles in the venous flow system to probe human diseases.

Staggered Multiple-PRF Ultrafast Color Doppler.

Color Doppler imaging is an established pulsed ultrasound technique to visualize blood flow non-invasively. High-frame-rate (ultrafast) color Doppler, by emissions of plane or circular wavefronts, allows severalfold increase in frame rates. Conventional and ultrafast color Doppler are both limited by the range-velocity dilemma, which may result in velocity folding (aliasing) for large depths and/or large velocities. We investigated multiple pulse-repetition-frequency (PRF) emissions arranged in a series of staggered intervals to remove aliasing in ultrafast color Doppler. Staggered PRF is an emission process where time delays between successive pulse transmissions change in an alternating way. We tested staggered dual- and triple-PRF ultrafast color Doppler, 1) in vitro in a spinning disc and a free jet flow, and 2) in vivo in a human left ventricle. The in vitro results showed that the Nyquist velocity could be extended to up to 6 times the conventional limit. We found coefficients of determination r(2) ≥ 0.98 between the de-aliased and ground-truth velocities. Consistent de-aliased Doppler images were also obtained in the human left heart. Our results demonstrate that staggered multiple-PRF ultrafast color Doppler is efficient for high-velocity high-frame-rate blood flow imaging. This is particularly relevant for new developments in ultrasound imaging relying on accurate velocity measurements.