Validation of ultrasound velocimetry and computational fluid dynamics for flow assessment in femoral artery stenotic disease.

Purpose: To investigate the accuracy of high-framerate echo particle image velocimetry (ePIV) and computational fluid dynamics (CFD) for determining velocity vectors in femoral bifurcation models through comparison with optical particle image velocimetry (oPIV). Approach: Separate femoral bifurcation models were built for oPIV and ePIV measurements of a non-stenosed (control) and a 75%-area stenosed common femoral artery. A flow loop was used to create triphasic pulsatile flow. In-plane velocity vectors were measured with oPIV and ePIV. Flow was simulated with CFD using boundary conditions from ePIV and additional duplex-ultrasound (DUS) measurements. Mean differences and 95%-limits of agreement (1.96*SD) of the velocity magnitudes in space and time were compared, and the similarity of vector complexity (VC) and time-averaged wall shear stress (TAWSS) was assessed. Results: Similar flow features were observed between modalities with velocities up to 110 and 330 cm/s in the control and the stenosed model, respectively. Relative to oPIV, ePIV and CFD-ePIV showed negligible mean differences in velocity (<3 cm/s), with limits of agreement of ±25 cm/s (control) and ±34 cm/s (stenosed). CFD-DUS overestimated velocities with limits of agreements of 13±40 and 16.1±55 cm/s for the control and stenosed model, respectively. VC showed good agreement, whereas TAWSS showed similar trends but with higher values for ePIV, CFD-DUS, and CFD-ePIV compared to oPIV. Conclusions: EPIV and CFD-ePIV can accurately measure complex flow features in the femoral bifurcation and around a stenosis. CFD-DUS showed larger deviations in velocities making it a less robust technique for hemodynamical assessment. The applied ePIV and CFD techniques enable two- and three-dimensional assessment of local hemodynamics with high spatiotemporal resolution and thereby overcome key limitations of current clinical modalities making them an attractive and cost-effective alternative for hemodynamical assessment in clinical practice.

A unifying Rayleigh-Plesset-type equation for bubbles in viscoelastic media.

Understanding the ultrasound pressure-driven dynamics of microbubbles confined in viscoelastic materials is relevant for multiple biomedical applications, ranging from contrast-enhanced ultrasound imaging to ultrasound-assisted drug delivery. The volumetric oscillations of spherical bubbles are analyzed using the Rayleigh-Plesset equation, which describes the conservation of mass and momentum in the surrounding medium. Several studies have considered an extension of the Rayleigh-Plesset equation for bubbles embedded into viscoelastic media, but these are restricted to a particular choice of constitutive model and/or to small deformations. Here, we derive a unifying equation applicable to bubbles in viscoelastic media with arbitrary complex moduli and that can account for large bubble deformations. To derive this equation, we borrow concepts from finite-strain theory. We validate our approach by comparing the result of our model to previously published results and extend it to show how microbubbles behave in arbitrary viscoelastic materials. In particular, we use our viscoelastic Rayleigh-Plesset model to compute the bubble dynamics in benchmarked viscoelastic liquids and solids.

An anthropomorphic thyroid phantom for ultrasound-guided radiofrequency ablation of nodules.

BACKGROUND: Needle-based procedures, such as fine needle aspiration and thermal ablation, are often applied for thyroid nodule diagnosis and therapeutic purposes, respectively. With blood vessels and nerves nearby, these procedures can pose risks in damaging surrounding critical structures. PURPOSE: The development and validation of innovative strategies to manage these risks require a test object with well-characterized physical properties. For this work, we focus on the application of ultrasound-guided thermal radiofrequency ablation. METHODS: We have developed a single-use anthropomorphic phantom mimicking the thyroid and surrounding anatomical and physiological structures that are relevant to ultrasound-guided thermal ablation. The phantom was composed of a mixture of polyacrylamide, water, and egg white extract and was cast using molds in multiple steps. The thermal, acoustical, and electrical characteristics were experimentally validated. The ablation zones were analyzed via non-destructive T2 -weighted magnetic resonance imaging scans utilizing the relaxometry changes of coagulated egg albumen, and the temperature distribution was monitored using an array of fiber Bragg grating sensors. RESULTS: The physical properties of the phantom were verified both on ultrasound as well as in terms of the phantom response to thermal ablation. The final temperature achieved (92°C), the median percentage of the nodule ablated (82.1%), the median volume ablated outside the nodule (0.8 mL), and the median number of critical structures affected (0) were quantified. CONCLUSION: An anthropomorphic phantom that can provide a realistic model for development and training in ultrasound-guided needle-based thermal interventions for thyroid nodules has been presented.

Ultrasound Particle Image Velocimetry to Investigate Potential Hemodynamic Causes of Limb Thrombosis After Endovascular Aneurysm Repair With the Anaconda Device.

PURPOSE: To identify potential hemodynamic predictors for limb thrombosis (LT) following endovascular aneurysm repair with the Anaconda endograft in a patient-specific phantom. MATERIALS AND METHODS: A thin-walled flow phantom, based on a patient's aortic anatomy and treated with an Anaconda endograft, that presented with a left-sided LT was fabricated. Contrast-enhanced ultrasound particle image velocimetry was performed to quantify time-resolved velocity fields. Measurements were performed in the same phantom with and without the Anaconda endograft, to investigate the impact of the endograft on the local flow fields. Hemodynamic parameters, namely vector complexity (VC) and residence time (RT), were calculated for both iliac arteries. RESULTS: In both limbs, the vector fields were mostly unidirectional during the peak systolic and end-systolic velocity phases before and after endograft placement. Local vortical structures and complex flow fields were observed at the diastolic and transitional flow phases. The average VC was higher (0.11) in the phantom with endograft, compared to the phantom without endograft (0.05). Notably, in both left and right iliac arteries, the anterior wall regions corresponded to a 2- and 4-fold increase in VC in the phantom with endograft, respectively. RT simulations showed values of 1.3 to 6 seconds in the phantom without endograft. A higher RT (up to 25 seconds) was observed in the phantom with endograft, in which the left iliac artery, with LT in follow-up, showed 2 fluid stasis regions. CONCLUSION: This in vitro study shows that unfavorable hemodynamics were present mostly in the limb that thrombosed during follow-up, with the highest VC and longest RT. These parameters might be valuable in predicting the occurrence of LT in the future. CLINICAL IMPACT: This in-vitro study aimed to identify potential hemodynamic predictors for limb thrombosis following EVAR using ultrasound particle image velocimetry (echoPIV) technique. It was shown that unfavorable hemodynamic norms were present mostly in the thrombosed limb. Owing to the in-vivo feasibility of the echoPIV, future efforts should focus on the evaluation of these hemodynamic norms in clinical trials. Thereafter, using echoPIV as a bedside technique in hospitals becomes more promising. Performing echoPIV in pre-op phase may provide valuable insights for surgeons to enhance treatment planning. EchoPIV is also applicable for follow-up sessions to evaluate treatment progress and avoid/predict complications.

Dependence of sonoporation efficiency on microbubble size: An in vitro monodisperse microbubble study.

Sonoporation is the process where intracellular drug delivery is facilitated by ultrasound-driven microbubble oscillations. Several mechanisms have been proposed to relate microbubble dynamics to sonoporation including shear and normal stress. The present work aims to gain insight into the role of microbubble size on sonoporation and thereby into the relevant mechanism(s) of sonoporation. To this end, we measured the sonoporation efficiency while varying microbubble size using monodisperse microbubble suspensions. Sonoporation experiments were performed in vitro on cell monolayers using a single ultrasound pulse with a fixed frequency of 1 MHz while the acoustic pressure amplitude and pulse length were varied at 250, 500, and 750 kPa, and 10, 100, and 1000 cycles, respectively. Sonoporation efficiency was quantified using flow cytometry by measuring the FITC-dextran (4 kDa and 2 MDa) fluorescence intensity in 10,000 cells per experiment to average out inherent variations in the bioresponse. Using ultra-high-speed imaging at 10 million frames per second, we demonstrate that the bubble oscillation amplitude is nearly independent of the equilibrium bubble radius at acoustic pressure amplitudes that induce sonoporation (≥ 500 kPa). However, we show that sonoporation efficiency is strongly dependent on the equilibrium bubble size and that under all explored driving conditions most efficiently induced by bubbles with a radius of 4.7 µm. Polydisperse microbubbles with a typical ultrasound contrast agent size distribution perform almost an order of magnitude lower in terms of sonoporation efficiency than the 4.7-µm bubbles. We elucidate that for our system shear stress is highly unlikely the mechanism of action. By contrast, we show that sonoporation efficiency correlates well with an estimate of the bubble-induced normal stress.

Buckling of lipidic ultrasound contrast agents under quasi-static load.

Collapse of lipidic ultrasound contrast agents under high-frequency compressive load has been historically interpreted by the vanishing of surface tension. By contrast, buckling of elastic shells is known to occur when costly compressible stress is released through bending. Through quasi-static compression experiments on lipidic shells, we analyse the buckling events in the framework of classical elastic buckling theory and deduce the mechanical characteristics of these shells. They are then compared with that obtained through acoustic characterization. This article is part of the theme issue 'Probing and dynamics of shock sensitive shells'.

Ultrasound imaging in thyroid nodule diagnosis, therapy, and follow-up: Current status and future trends.

Ultrasound, the primary imaging modality in thyroid nodule management, suffers from drawbacks including: high inter- and intra-observer variability, limited field-of-view and limited functional imaging. Developments in ultrasound technologies are taking place to overcome these limitations, including three-dimensional-Doppler, -elastography, -nodule characteristics-extraction, and novel machine-learning algorithms. For thyroid ablative treatments and biopsies, perioperative use of three-dimensional ultrasound opens a new field of research. This review provides an overview of the current and future applications of ultrasound, and discusses the potential of new developments and trends that may improve the diagnosis, therapy, and follow-up of thyroid nodules.

Computational Fluid Dynamics for the Prediction of Endograft Thrombosis in the Superficial Femoral Artery.

PURPOSE: Contemporary diagnostic modalities, including contrast-enhanced computed tomography (CTA) and duplex ultrasound, have been insufficiently able to predict endograft thrombosis. This study introduces an implementation of image-based computational fluid dynamics (CFD), by exemplification with 4 patients treated with an endograft for occlusive disease of the superficial femoral artery (SFA). The potential of personalized CFD for predicting endograft thrombosis is investigated. MATERIALS AND METHODS: Four patients treated with endografts for an occluded SFA were retrospectively included. CFD simulations, based on CTA and duplex ultrasound, were compared for patients with and without endograft thrombosis to investigate potential flow-related causes of endograft thrombosis. Time-averaged wall shear stress (TAWSS) was computed, which highlights areas of prolonged residence times of coagulation factors in the graft. RESULTS: CFD simulations demonstrated normal TAWSS (>0.4 Pa) in the SFA for cases 1 and 2, but low levels of TAWSS (<0.4 Pa) in cases 3 and 4, respectively. Primary patency was achieved in cases 1 and 2 for over 2 year follow-up. Cases 3 and 4 were complicated by recurrent endograft thrombosis. CONCLUSION: The presence of a low TAWSS was associated with recurrent endograft thrombosis in subjects with otherwise normal anatomic and ultrasound assessment and a good distal run-off.

High-frame-rate contrast-enhanced ultrasound particle image velocimetry in patients with a stented superficial femoral artery: a feasibility study.

BACKGROUND: Local blood flow affects vascular disease and outcomes of endovascular treatment, but quantifying it is challenging, especially inside stents. We assessed the feasibility of blood flow quantification in native and stented femoral arteries, using high-frame-rate (HFR) contrast-enhanced ultrasound (CEUS) particle image velocimetry (PIV), also known as echoPIV. METHODS: Twenty-one patients with peripheral arterial disease, recently treated with a stent in the femoral artery, were included. HFR CEUS measurements were performed in the native femoral artery and at the inflow and outflow of the stent. Two-dimensional blood flow was quantified through PIV analysis. EchoPIV recordings were visually assessed by five observers and categorised as optimal, partial, or unfeasible. To evaluate image quality and tracking performance, contrast-to-tissue ratio (CTR) and vector correlation were calculated, respectively. RESULTS: Fifty-eight locations were measured and blood flow quantification was established in 49 of them (84%). Results were optimal for 17/58 recordings (29%) and partial for 32 recordings (55%) due to loss of correlation (5/32; 16%), short vessel segment (8/32; 25%), loss of contrast (14/32; 44%), and/or shadows (18/32; 56%). In the remaining 9/58 measurements (16%) no meaningful flow information was visualised. Overall, CTR and vector correlation were lower during diastole. CTR and vector correlation were not different between stented and native vessel segments, except for a higher native CTR at the inflow during systole (p = 0.037). CONCLUSIONS: Blood flow quantification is feasible in untreated and stented femoral arteries using echoPIV. Limitations remain, however, none of them related to the presence of the stent. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04934501 (retrospectively registered).

Time-resolved absolute radius estimation of vibrating contrast microbubbles using an acoustical camera.

Ultrasound (US) contrast agents consist of microbubbles ranging from 1 to 10 µm in size. The acoustical response of individual microbubbles can be studied with high-frame-rate optics or an "acoustical camera" (AC). The AC measures the relative microbubble oscillation while the optical camera measures the absolute oscillation. In this article, the capabilities of the AC are extended to measure the absolute oscillations. In the AC setup, microbubbles are insonified with a high- (25 MHz) and low-frequency US wave (1-2.5 MHz). Other than the amplitude modulation (AM) from the relative size change of the microbubble (employed in Renaud, Bosch, van der Steen, and de Jong (2012a). "An 'acoustical camera' for in vitro characterization of contrast agent microbubble vibrations," Appl. Phys. Lett. 100(10), 101911, the high-frequency response from individual vibrating microbubbles contains a phase modulation (PM) from the microbubble wall displacement, which is the extension described here. The ratio of PM and AM is used to determine the absolute radius, R0. To test this sizing, the size distributions of two monodisperse microbubble populations ( R = 2.1 and 3.5 µm) acquired with the AC were matched to the distribution acquired with a Coulter counter. As a result of measuring the absolute size of the microbubbles, this "extended AC" can capture the full radial dynamics of single freely floating microbubbles with a throughput of hundreds of microbubbles per hour.

Irrigant flow in the root canal during ultrasonic activation: A numerical fluid-structure interaction model and its validation.

AIM: The aim of the study was (a) to develop a three-dimensional numerical model combining the oscillation of a tapered ultrasonic file and the induced irrigant flow along with their two-way interaction in the confinement of a root canal. (b) To validate this model through comparison with experiments and theoretical (analytical) solutions of the flow. METHODOLOGY: Two partial numerical models, one for the oscillation of the ultrasonic file and another one for the irrigant flow inside the root canal around the file, were created and coupled in order to take into account the two-way coupled fluid-structure interaction. Simulations were carried out for ultrasonic K-files and for smooth wires driven at four different amplitudes in air or inside an irrigant-filled straight root canal. The oscillation pattern of the K-files was determined experimentally by Scanning Laser Vibrometry, and the flow pattern inside an artificial root canal was analysed using high-speed imaging together with Particle Image Velocimetry. Analytical solutions were obtained from an earlier study. Numerical, experimental and analytical results were compared to assess the validity of the model. RESULTS: The comparison of the oscillation amplitude and node location of the ultrasonic files and of the irrigant flow field showed a close agreement between the simulations, experiments and theoretical solutions. CONCLUSIONS: The model is able to predict reliably the file oscillation and irrigant flow inside root canals during ultrasonic activation under similar conditions.

Blood Flow Quantification with High-Frame-Rate, Contrast-Enhanced Ultrasound Velocimetry in Stented Aortoiliac Arteries: In Vivo Feasibility.

Local flow patterns influence stent patency, while blood flow quantification in stents is challenging. The aim of this study was to investigate the feasibility of 2-D blood flow quantification using high-frame-rate, contrast-enhanced ultrasound (HFR-CEUS) and particle image velocimetry (PIV), or echoPIV, in patients with aortoiliac stents. HFR-CEUS measurements were performed at 129 locations in 62 patients. Two-dimensional blood flow velocity fields were obtained using echoPIV. Visual inspection was performed by five observers to evaluate feasibility. The contrast-to-background ratio and average vector correlation were calculated and compared between stented and native vessel segments. Flow quantification with echoPIV was feasible in 128 of 129 locations (99%), with optimal quantification in 40 of 129 locations (31%). Partial quantification was achieved in 88 of 129 locations (68%), where one or multiple limiting issues occurred (not related to the stent) including loss of correlation during systole (57/129), short vessel segments (20/129), loss of contrast during diastole (20/129) and shadow regions (20/129). The contrast-to-background ratio and vector correlation were lower downstream in the imaged blood vessel, independent of the location of the stent. In conclusion, echoPIV was feasible in stents placed in the aortoiliac region, and the stents did not adversely affect flow tracking.

Resonance behavior of a compliant piezo-driven inkjet channel with an entrained microbubble.

Microbubbles entrained in a piezo-driven drop-on-demand printhead disturb the acoustics of the microfluidic ink channel and, thereby, the jetting behavior. Here, the resonance behavior of an ink channel as a function of the microbubble size and number of bubbles is studied through theoretical modeling and experiments. The system is modeled as a set of two coupled harmonic oscillators: one corresponds to the compliant ink channel and the other corresponds to the microbubble. The predicted and measured eigenfrequencies are in excellent agreement. It was found that the resonance frequency is independent of the bubble size as long as the compliance of the bubble dominates over that of the piezo actuator. An accurate description of the eigenfrequency of the coupled system requires the inclusion of the increased inertance of the entrained microbubble due to confinement. It is shown that the inertance of a confined bubble can be accurately obtained by using a simple potential flow approach. The model is further validated by the excellent agreement between the modeled and measured microbubble resonance curves. The present work, therefore, provides physical insight into the coupled dynamics of a compliant ink channel with an entrained microbubble.

The response of dual-species bacterial biofilm to 2% and 5% NaOCl mixed with etidronic acid: A laboratory real-time evaluation using optical coherence tomography.

AIM: The addition of etidronic acid (HEDP) to sodium hypochlorite (NaOCl) could increase the antibiofilm potency of the irrigant, whilst maintaining the benefits of continuous chelation. Studies conducted so far have shown that mixing HEDP with NaOCl solutions of relatively low concentration does not compromise the antibiofilm efficacy of the irrigant. However, the working lifespan of NaOCl may decrease resulting in a reduction of its antibiofilm efficacy over time (efficiency). In this regard, continuous irrigant replenishment needs to be examined. This study investigated the response of a dual-species biofilm when challenged with 2% and 5% NaOCl mixed with HEDP for a prolonged timespan and under steady laminar flow. METHODOLOGY: Dual-species biofilms comprised of Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 were grown on human dentine discs in a constant depth film fermenter (CDFF) for 96 h. Biofilms were treated with 2% and 5% NaOCl, alone or mixed with HEDP. Irrigants were applied under steady laminar flow for 8 min. Biofilm response was evaluated by means of optical coherence tomography (OCT). Biofilm removal, biofilm disruption, rate of biofilm loss and disruption as well as bubble formation were assessed. One-way anova, Wilcoxon's signed-rank test and Kruskal-Wallis H test were performed for statistical analysis of the data. The level of significance was set at a ≤.05. RESULTS: Increasing NaOCl concentration resulted in increased biofilm removal and disruption, higher rate of biofilm loss and disruption and increased bubble formation. Mixing HEDP with NaOCl caused a delay in the antibiofilm action of the latter, without compromising its antibiofilm efficacy. CONCLUSIONS: NaOCl concentration dictates the biofilm response irrespective of the presence of HEDP. The addition of HEDP resulted in a delay in the antibiofilm action of NaOCl. This delay affects the efficiency, but not the efficacy of the irrigant over time.

Super-Resolved Microbubble Localization in Single-Channel Ultrasound RF Signals Using Deep Learning.

Recently, super-resolution ultrasound imaging with ultrasound localization microscopy (ULM) has received much attention. However, ULM relies on low concentrations of microbubbles in the blood vessels, ultimately resulting in long acquisition times. Here, we present an alternative super-resolution approach, based on direct deconvolution of single-channel ultrasound radio-frequency (RF) signals with a one-dimensional dilated convolutional neural network (CNN). This work focuses on low-frequency ultrasound (1.7 MHz) for deep imaging (10 cm) of a dense cloud of monodisperse microbubbles (up to 1000 microbubbles in the measurement volume, corresponding to an average echo overlap of 94%). Data are generated with a simulator that uses a large range of acoustic pressures (5-250 kPa) and captures the full, nonlinear response of resonant, lipid-coated microbubbles. The network is trained with a novel dual-loss function, which features elements of both a classification loss and a regression loss and improves the detection-localization characteristics of the output. Whereas imposing a localization tolerance of 0 yields poor detection metrics, imposing a localization tolerance corresponding to 4% of the wavelength yields a precision and recall of both 0.90. Furthermore, the detection improves with increasing acoustic pressure and deteriorates with increasing microbubble density. The potential of the presented approach to super-resolution ultrasound imaging is demonstrated with a delay-and-sum reconstruction with deconvolved element data. The resulting image shows an order-of-magnitude gain in axial resolution compared to a delay-and-sum reconstruction with unprocessed element data.

US Velocimetry in Participants with Aortoiliac Occlusive Disease.

Background The accurate quantification of blood flow in aortoiliac arteries is challenging but clinically relevant because local flow patterns can influence atherosclerotic disease. Purpose To investigate the feasibility and clinical application of two-dimensional blood flow quantification using high-frame-rate contrast-enhanced US (HFR-CEUS) and particle image velocimetry (PIV), or US velocimetry, in participants with aortoiliac stenosis. Materials and Methods In this prospective study, participants with a recently diagnosed aortoiliac stenosis underwent HFR-CEUS measurements of the pre- and poststenotic vessel segments (August 2018 to July 2019). Two-dimensional quantification of blood flow was achieved by performing PIV analysis, which was based on pairwise cross-correlation of the HFR-CEUS images. Visual inspection of the entire data set was performed by five observers to evaluate the ability of the technique to enable adequate visualization of blood flow. The contrast-to-background ratio and average vector correlation were calculated. In two participants who showed flow disturbances, the flow complexity and vorticity were calculated. Results Thirty-five participants (median age, 67 years; age range, 56-84 years; 22 men) were included. Visual scoring showed that flow quantification was achieved in 41 of 42 locations. In 25 locations, one or multiple issues occurred that limited optimal flow quantification, including loss of correlation during systole (n = 12), shadow regions (n = 8), a short vessel segment in the image plane (n = 7), and loss of contrast during diastole (n = 5). In the remaining 16 locations, optimal quantification was achieved. The contrast-to-background ratio was higher during systole than during diastole (11.0 ± 2.9 vs 6.9 ± 3.4, respectively; P < .001), whereas the vector correlation was lower (0.58 ± 0.21 vs 0.47 ± 0.13; P < .001). The flow complexity and vorticity were high in regions with disturbed flow. Conclusion Blood flow quantification with US velocimetry is feasible in patients with an aortoiliac stenosis, but several challenges must be overcome before implementation into clinical practice. Clinical trial registration no. NTR6980 © RSNA, 2021 Online supplemental material is available for this article.

Hemodynamic Comparison of Stent-Grafts for the Treatment of Aortoiliac Occlusive Disease.

PURPOSE: To compare the flow patterns and hemodynamics of the AFX stent-graft and the covered endovascular reconstruction of aortic bifurcation (CERAB) configuration using laser particle image velocimetry (PIV) experiments. MATERIALS AND METHODS: Two anatomically realistic aortoiliac phantoms were constructed using polydimethylsiloxane polymer. An AFX stent-graft with a transparent cover made with a new method was inserted into one phantom. A CERAB configuration using Atrium's Avanta V12 with transparent covers made with a previously established method was inserted into the other phantom, both modified stent-grafts were suitable for laser PIV, enabling visualization of the flow fields and quantification of time average wall shear stress (TAWSS), oscillatory shear index (OSI), and relative residence time (RRT). RESULTS: Disturbed flow was observed at the bifurcation region of the AFX, especially at the end systolic velocity (ESV) time-point where recirculation was noticeable due to vortical flow. In contrast, predominantly unidirectional flow was observed at the CERAB bifurcation. These observations were confirmed by the quantified hemodynamic results from PIV analysis where mean TAWSS of 0.078 Pa (range: 0.009-0.242 Pa) was significantly lower in AFX as compared with 0.229 Pa (range: 0.013-0.906 Pa) for CERAB (p<0.001). Mean OSI of 0.318 (range: 0.123-0.496) in AFX was significantly higher than 0.252 (range: 0.055-0.472) in CERAB (p<0.001). Likewise, mean RRT of 180 Pa-1 (range: 9-3603 Pa-1) in AFX was also significantly higher than 88 Pa-1 (range: 2-840 Pa-1) in CERAB (p=0.0086). CONCLUSION: In this in vitro study, the flow pattern of a modified AFX stent-graft was found to be more disturbed especially at the end systolic phase, its hemodynamic outcomes less desirable than CERAB configuration. CLINICAL RELEVANCE: While the AFX stent-graft has an advantage over the CERAB configuration in eliminating radial mismatch, and maintaining the anatomical bifurcation for future endovascular intervention, this in vitro study revealed that the associated lower TAWSS, higher OSI and RRT may predispose to thrombosis and are, thus, less desirable as compared to a CERAB configuration. Further investigation is warranted to confirm whether these findings translate into the clinical setting.

Multi-timescale Microscopy Methods for the Characterization of Fluorescently-labeled Microbubbles for Ultrasound-Triggered Drug Release.

Microbubble contrast agents hold great promise for drug delivery applications with ultrasound. Encapsulating drugs in nanoparticles reduces systemic toxicity and increases circulation time of the drugs. In a novel approach to microbubble-assisted drug delivery, nanoparticles are incorporated in or on microbubble shells, enabling local and triggered release of the nanoparticle payload with ultrasound. A thorough understanding of the release mechanisms within the vast ultrasound parameter space is crucial for efficient and controlled release. This set of presented protocols is applicable to microbubbles with a shell containing a fluorescent label. Here, the focus is on microbubbles loaded with poly(2-ethyl-butyl cyanoacrylate) polymeric nanoparticles, doped with a modified Nile Red dye. The particles are fixed within a denatured casein shell. The microbubbles are produced by vigorous stirring, forming a dispersion of perfluoropropane gas in the liquid phase containing casein and nanoparticles, after which the microbubble shell self-assembles. A variety of microscopy techniques are needed to characterize the nanoparticle-stabilized microbubbles at all relevant timescales of the nanoparticle release process. Fluorescence of the nanoparticles enables confocal imaging of single microbubbles, revealing the particle distribution within the shell. In vitro ultra-high-speed imaging using bright-field microscopy at 10 million frames per second provides insight into the bubble dynamics in response to ultrasound insonation. Finally, nanoparticle release from the bubble shell is best visualized by means of fluorescence microscopy, performed at 500,000 frames per second. To characterize drug delivery in vivo, the triggered release of nanoparticles within the vasculature and their extravasation beyond the endothelial layer is studied using intravital microscopy in tumors implanted in dorsal skinfold window chambers, over a timescale of several minutes. The combination of these complementary characterization techniques provides unique insight into the behavior of microbubbles and their payload release at a range of time and length scales, both in vitro and in vivo.

Blood Flow Quantification in Peripheral Arterial Disease: Emerging Diagnostic Techniques in Vascular Surgery.

The assessment of local blood flow patterns in patients with peripheral arterial disease is clinically relevant, since these patterns are related to atherosclerotic disease progression and loss of patency in stents placed in peripheral arteries, through mechanisms such as recirculating flow and low wall shear stress (WSS). However, imaging of vascular flow in these patients is technically challenging due to the often complex flow patterns that occur near atherosclerotic lesions. While several flow quantification techniques have been developed that could improve the outcomes of vascular interventions, accurate 2D or 3D blood flow quantification is not yet used in clinical practice. This article provides an overview of several important topics that concern the quantification of blood flow in patients with peripheral arterial disease. The hemodynamic mechanisms involved in the development of atherosclerosis and the current clinical practice in the diagnosis of this disease are discussed, showing the unmet need for improved and validated flow quantification techniques in daily clinical practice. This discussion is followed by a showcase of state-of-the-art blood flow quantification techniques and how these could be used before, during and after treatment of stenotic lesions to improve clinical outcomes. These techniques include novel ultrasound-based methods, Phase-Contrast Magnetic Resonance Imaging (PC-MRI) and Computational Fluid Dynamics (CFD). The last section discusses future perspectives, with advanced (hybrid) imaging techniques and artificial intelligence, including the implementation of these techniques in clinical practice.