Contrast-enhanced Ultrasound Measurements Research Articles

The Size Differences of Breast Cancer and Benign Tumors Measured by Two-Dimensional Ultrasound and Contrast-Enhanced Ultrasound.

Ultrasound (US) imaging has been observed to underestimate tumor size in clinical practice. This study aims to compare the size measurements of breast cancer and benign tumors using two-dimensional ultrasound (2DUS) and contrast-enhanced ultrasound (CEUS). The study included 42 clinically confirmed breast cancer and 47 benign breast tumors. Two experienced physicians independently measured the maximal longitudinal and transverse diameters of the masses in 2DUS and CEUS. All analyses were performed in R (4.2.2) and GraphPad Prism 6. The maximal longitudinal and transverse diameters of breast cancer measured by CEUS were 26.61 ± 0.21% and 26.24 ± 0.19% larger compared with 2DUS, and benign breast tumors had an 11.74 ± 0.21% and 11.06 ± 0.14% increase in size compared with 2DUS. The area under the curve (AUC) of the receiver operating characteristic curve (ROC) for the difference between 2DUS and CEUS was 0.870 for longitudinal diameters (95% CI: 0.795-0.945, sensitivity 0.842, specificity 0.783, threshold value 0.215), and 0.863 for transverse diameters (95% CI: 0.785-0.942, sensitivity 0.667, specificity 0.936, threshold value 0.203). The size measurements of both breast cancer and benign tumors were larger in CEUS compared with 2DUS, with CEUS measurements of breast cancer being more pronounced than those of benign breast tumors. These findings suggest that CEUS may provide a more precise assessment of tumor size, which is crucial for determining optimal treatment strategies and improving patient outcomes in breast cancer management.

ASSESSMENT OF ABNORMAL SKELETAL MUSCLE PERFUSION BY CONTRAST-ENHANCED ULTRASOUND WITH PARAMETRIC IMAGING IN RATS AFTER SEVERE INJURY, HEMORRHAGIC SHOCK, AND WHOLE BLOOD RESUSCITATION.

Background: After severe injury, optical measures of microvascular blood flow (MBF) decrease and do not normalize with resuscitation to normal blood pressure. These changes are associated with organ dysfunction, coagulopathy, and death. However, the pathophysiology is not well understood. Several possible pathways could also contribute to the development of trauma-induced coagulopathy (TIC). A small-animal model of trauma-related MBF derangement that persists after resuscitation and includes TIC would facilitate further study. Parametric contrast-enhanced ultrasound (CEUS) is particularly advantageous in this setting, because it noninvasively assesses MBF in large, deep vascular beds. We sought to develop such a model, measuring MBF with CEUS. Methods: Sixteen male Sprague-Dawley rats were anesthetized, ventilated, and cannulated. Rats were subjected to either no injury (sham group) or a standardized polytrauma and pressure-targeted arterial catheter hemorrhage with subsequent whole blood resuscitation (trauma group). At prespecified time points, CEUS measurements of uninjured quadriceps muscle, viscoelastic blood clot strength, and complete blood counts were taken. Results: After resuscitation, blood pressure normalized, but MBF decreased and remained low for the rest of the protocol. This was primarily driven by a decrease in blood volume with a relative sparing of blood velocity. Viscoelastic blood clot strength and platelet count also decreased and remained low throughout the protocol. Conclusion: We present a rat model of MBF derangement in uninjured skeletal muscle and coagulopathy after polytrauma that persists after resuscitation with whole blood to normal macrohemodynamics. Parametric CEUS analysis shows that this change is primarily due to microvascular obstruction. This platform can be used to develop a deeper understanding of this important process.

Real-Time Elastography versus Shear Wave Elastography on Evaluating the Timely Radiofrequency Ablation Effect of Rabbit Liver: A Preliminary Experimental Study

to evaluate and monitor the timely thermal ablation changes of rabbit liver by using two elastographic methods-real-time elastography (RTE) and shear wave elastography (SWE)-as compared to contrast-enhanced ultrasound (CEUS) and physical specimens. 20 ablation zones were created in the livers of 20 rabbits using radiofrequency ablation (RFA). After the ablation, RTE and SWE were used to measure the elastic properties of the twenty ablation zones. The consistency of efficacy evaluation for RTE and SWE measurements was analyzed using the Bland-Altman test. The areas of the thermal ablation zones were also measured and compared according to the images provided by RTE, SWE, CEUS, and gross physical specimen measurement. RTE and SWE could clearly display the shape of RFA ablation zones within one hour after the ablation. The average elasticity ratio for the ablation zone measured by RTE was 3.41 ± 0.67 (2.23-4.76); the average elasticity value measured by SWE was 50.7 ± 11.3 kPa (33.2-70.4 kPa). The mean areas of the ablation zones measured with RTE, SWE, gross specimen, and CEUS were 1.089 ± 0.199 cm2, 1.059 ± 0.201 cm2, 1.081 ± 0.201 cm2, and 3.091 ± 0.591 cm2, respectively. The Bland-Altman test showed that RTE and SWE have great consistency. Area measurements by CEUS were significantly larger than those of the other three methods (p < 0.05). RTE and SWE are both able to accurately confirm the range of ablation zones shortly after the ablation for rabbit livers.

Contrast-Enhanced Ultrasound in the Differentiation between the Most Common Benign Parotid Gland Tumors: A Systematic Review and Meta-Analysis.

Recently, contrast-enhanced ultrasound (CEUS) has become a promising tool in distinguishing benign from malignant parotid gland tumors. However, its usefulness in differentiating various benign parotid tumors has not been determined so far. This study aimed to systematically review the literature to determine the utility of CEUS in the preoperative differentiation between pleomorphic adenomas (PAs) and Warthin's tumors (WTs) of the parotid gland. PubMed, Embase, and Cochrane were searched for English-language articles published until 21 July 2022. Fifteen studies were included. On CEUS examination, a significantly greater percentage of PAs displayed heterogeneous enhancement texture compared to WTs. Contrarily, the enhanced lesion size, the enhancement margin, and the presence of the enhancement rim did not differ significantly between the entities. Significantly longer normalized mean transit time (nMTT) and time to peak (TTP) were observed in PAs. Contrarily, the mean values of area under the curve (AUC) and time from peak to one half (TPH) were significantly higher for WTs. Due to the considerable overlap among the qualitative CEUS characteristics of PAs and WTs, the reproducible, investigator-independent quantitative CEUS measurements have a greater potential to distinguish PAs from WTs, which might influence the selection of an appropriate management strategy.

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

BackgroundLocal 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.MethodsTwenty-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.ResultsFifty-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).ConclusionsBlood 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 registrationClinicalTrials.gov, NCT04934501 (retrospectively registered).

Ultrasound Imaging of Pancreatic Perfusion Dynamics Predicts Therapeutic Prevention of Diabetes in Preclinical Models of Type 1 Diabetes.

In type 1 diabetes (T1D), immune-cell infiltration into islets of Langerhans (insulitis) and β-cell decline occur years before diabetes presents. There is a lack of validated clinical approaches for detecting insulitis and β-cell decline, to diagnose eventual diabetes and monitor the efficacy of therapeutic interventions. We previously determined that contrast-enhanced ultrasound measurements of pancreas perfusion dynamics predict disease progression in T1D pre-clinical models. Here, we test whether these measurements predict therapeutic prevention of T1D. We performed destruction-reperfusion measurements with size-isolated microbubbles in non-obese diabetic (NOD)-severe combined immunodeficiency (SCID) mice receiving an adoptive transfer of diabetogenic splenocytes. Mice received vehicle control or the following treatments: (i) anti-CD3 to block T-cell activation; (ii) anti-CD4 to deplete CD4+ T cells; (iii) verapamil to reduce β-cell apoptosis; or (iv) tauroursodeoxycholic acid (TUDCA) to reduce β-cell endoplasmic reticulum stress. We compared measurements of pancreas perfusion dynamics with subsequent progression to diabetes. Anti-CD3, anti-CD4, and verapamil delayed diabetes development. Blood flow dynamics was significantly altered in treated mice with delayed/absent diabetes development compared with untreated mice. Conversely, blood flow dynamics in treated mice with unchanged diabetes development was similar to that in untreated mice. Thus, measurement of pancreas perfusion dynamics predicts the successful prevention of diabetes. This strategy may provide a clinically deployable predictive marker for therapeutic prevention in asymptomatic T1D.

Welfare Assessment on Healthy and Tumor-Bearing Mice after Repeated Ultrasound Imaging

Introduction: Ultrasound (US) imaging enables tissue visualization in high spatial resolution with short examination times. Thus, it is often applied in preclinical research. Diagnostic US, including contrast-enhanced US (CEUS), is considered to be well-tolerated by laboratory animals although no systematic study has been performed to confirm this claim. Therefore, the aim of this study was to screen for possible effects of US and CEUS examinations on welfare of healthy mice. Additionally, the potential influence of CEUS and molecular CEUS on well-being and therapy response to regorafenib was investigated in breast cancer-bearing mice. Material and Methods: Forty healthy Balb/c mice were randomly assigned for examination with US or CEUS (3×/week) for 4 weeks. Untreated healthy mice and mice receiving only isoflurane anesthesia served as controls (n = 10/group). Ninety-four 4T1 tumor-bearing Balb/c mice were allocated randomly to the following groups: no imaging, isoflurane anesthesia, CEUS, and molecular CEUS. They either received 10 mg/kg regorafenib or vehicle solution daily by oral gavage. Animals were examined three times within 2 weeks. CEUS measurements were performed using phospholipid microbubbles, and phospholipid microbubbles targeting the vascular endothelial growth factor receptor-2 were applied for molecular CEUS. Welfare evaluation was performed by daily observational score sheets, measuring the heart rate, Rotarod performance, and fecal corticosterone metabolites twice a week. On the last day, pathological changes in serum corticosterone concentrations, hemograms, and organ weights were obtained. Moreover, a potential influence of isoflurane anesthesia, CEUS, and molecular CEUS on regorafenib response in tumor-bearing mice was examined. Analysis of variance and Dunnett’s post hoc test were performed as statistical analyses. Results: Severity parameters were not altered after repeated US and CEUS examinations of healthy mice, but spleen sizes were significantly lower after isoflurane anesthesia. In tumor-bearing mice, no effect on animal welfare after repeated CEUS and molecular CEUS could be observed. However, leukocyte counts and spleen weights of tumor-bearing mice were significantly lower in animals examined with CEUS and molecular CEUS compared to the control groups. This effect was not visible in regorafenib-treated animals. Conclusions: Repeated US and (molecular) CEUS have no detectable impact on animal welfare in healthy and tumor-bearing mice. However, CEUS and molecular CEUS in combination with isoflurane anesthesia might attenuate immunological processes in tumor-bearing animals and may consequently affect responses to antitumor therapy.

Super-resolution ultrasound imaging : Methods and applications

The microvasculature plays an important role in many pathologic conditions but cannot be characterized in high resolution via conventional ultrasound methods. Doppler-based techniques, contrast-enhanced sonography as well as dynamic contrast-enhanced computed tomography (CT) and magnetic resonance imaging (MRI) are commonly used to characterize tissue vascularization. However, these techniques cannot visualize the microvasculature adequately. Ultrasound localization microscopy (ULM) consists of contrast-enhanced ultrasound measurements in combination with acomplex post-processing algorithm which detects microbubbles with high precision. The vasculature can then be visualized by accumulating the microbubble positions in afinal image. Compared to conventional ultrasound techniques, ULM improves the image resolution by afactor of more than10. This currently results in resolutions down to 10 µm and allows, therefore, the visualization of capillaries and the assessment of their perfusion. Also, this does not lead to areduction of the penetration depth or the signal-to-noise ratio (SNR). The method enables the visualization of vascular structures in unsurpassed detail and has the potential to offer new possibilities for the diagnosis of various diseases and for gaining insights into physiological processes. However, ULM is not commercially available yet but is intensely being tested in clinical studies. ULM could potentially be applied to all fields in which the vasculature is relevant. Current fields of application include oncology, nephrology, and neurological research.

The Association Between Contrast-Enhanced Ultrasound and Near-Infrared Spectroscopy-Derived Measures of Calf Muscle Microvascular Responsiveness in Older Adults

Contrast-enhanced ultrasound (CEUS) measures of post-occlusion skeletal muscle microvascular responsiveness demonstrate the microvascular dysfunction associated with ageing and age-related disease. However, the accessibility of CEUS is limited by the need for intravenous administration of ultrasound contrast agents and sophisticated imaging analysis. Alternative methods are required for the broader assessment ofmicrovascular dysfunction in research and clinical settings. Therefore, we aimed to evaluate the level of association and agreement between CEUS and near-infrared spectroscopy (NIRS)-derived measures of post-occlusion skeletal muscle microvascular responsiveness in older adults. During supine rest, participants (n=15, 67±11 years) underwent 5 minutes of thigh cuff-occlusion (200 mmHg). Post-occlusion CEUS measures of calf muscle microvascular responsiveness were made, including time to 95% peak acoustic intensity (TTP95 AI) and the rate of rise (slope AI). Simultaneous measures, including time to 95% peak oxygenated haemoglobin (TTP95 O2Hb) and slope O2Hb, were made using continuous-wave NIRS in the same muscle region. There were strong correlations between TTP95 measures derived from CEUS and NIRS (r=0.834, p=<0.001) and the corresponding measures of slope (r=0.735, p=0.004). The limits of agreement demonstrated by Bland Altman plot analyses for CEUS and NIRS-derived measures of TTP95 (-9.67-1.98 s) and slope (-1.29-5.23%. s-1) were smaller than the minimum differences expected in people with microvascular dysfunction. The strong correlations and level of agreement in the present study support the use of NIRS as a non-invasive, portable and cost-effective method for assessing post-occlusion skeletal muscle microvascular responsiveness in older adults.

Multiparametric ultrasound imaging for the assessment of normal versus steatotic livers

Liver disease is increasing in prevalence across the globe. We present here a multiparametric ultrasound (mpUS) imaging approach for assessing nonalcoholic fatty liver disease (NALFD). This study was performed using rats (N = 21) that were fed either a control or methionine and choline deficient (MCD) diet. A mpUS imaging approach that includes H-scan ultrasound (US), shear wave elastography, and contrast-enhanced US measurements were then performed at 0 (baseline), 2, and 6 weeks. Thereafter, animals were euthanized and livers excised for histological processing. A support vector machine (SVM) was used to find a decision plane that classifies normal and fatty liver conditions. In vivo mpUS results from control and MCD diet fed animals reveal that all mpUS measures were different at week 6 (P < 0.05). Principal component analysis (PCA) showed that the H-scan US data contributed the highest percentage to the classification among the mpUS measurements. The SVM resulted in 100% accuracy for classification of normal and high fat livers and 92% accuracy for classification of normal, low fat, and high fat livers. Histology findings found considerable steatosis in the MCD diet fed animals. This study suggests that mpUS examinations have the potential to provide a comprehensive estimation of the main components of early stage NAFLD.

Contrast-Enhanced Brain Ultrasound Perfusion Metrics in the EXTra-Uterine Environment for Neonatal Development (EXTEND): Correlation With Hemodynamic Parameters.

Contrast-enhanced ultrasound (CEUS) can provide quantitative perfusion metrics and may be useful to detect cerebral pathology in neonates and premature infants, particularly in extrauterine environments. The effect of hemodynamics on cerebral perfusion metrics is unknown, which limits the clinical application of this technology. We aimed to determine associations between systemic hemodynamics and concurrently measured brain perfusion parameters in an animal model of extrauterine support. Nine fetal lambs were transferred to an extrauterine support device. Lumason® ultrasound contrast (0.1-0.3 ml) was administered via the umbilical vein and 90-second cine clips were obtained. Time-intensity-curves (TICs) were generated and time-dependent and area-under-curve (AUC) parameters were derived. Associations between brain perfusion metrics and hemodynamics including heart rate (HR) and mean arterial pressure (MAP) were evaluated by multilevel linear mixed-effects models. Eighty-six ultrasound examinations were performed and 72 examinations were quantifiable. Time-dependent measurements were independent of all hemodynamic parameters (all p ≥.05). Oxygen delivery and mean blood flow were correlated with AUC measurements (all p ≤.01). Physiologic HR and MAP were not correlated with any measurements (all p ≥.05). Detected aberrations in time-dependent CEUS measurements are not correlated with hemodynamic parameters and are thought to reflect the changes in cerebral blood flow, thus providing a promising tool for evaluation of brain perfusion. CEUS brain perfusion parameters are not correlated with physiologic HR and MAP, but AUC-dependent measurements are correlated with oxygen delivery and blood flow, suggesting that CEUS offers additional value over standard monitoring. Overall, these findings enhance the applicability of this technology.

Application of contrast-enhanced ultrasound-assisted superficial inferior epigastric artery perforator flap to repair oral and maxillofacial defects

To investigate the application value of contrast-enhanced ultrasound (CEUS) technique to assist the repair of oral and maxillofacial defects by superficial inferior epigastric artery perforator flap. Sixteen oral cancer patients, 10 males and 6 females, who were to undergo superficial inferior epigastric artery perforator flap repair between June 2018 and February 2020, were selected, with an average age of 55.8 years (range, 24-77 years). There were 13 cases of squamous cell carcinoma, 2 cases of adenoid cystic carcinoma, and 1 case of mucinous epidermis-like carcinoma. The color Doppler ultrasound (CDUS) and CEUS were used to screen the superficial inferior epigastric artery, assisted in the design of the flap, and compared it with the actual intraoperative exploration. The sensitivity, specificity, positive predictive value, and negative predictive value of CEUS and CDUS examinations were analyzed. Fourteen of 16 patients were repaired with superficial inferior epigastric artery perforator flap, and 2 patients were repaired with superficial iliac artery flap because the source artery was not found. After surgery, regular follow-up was performed to check for disease recurrence and metastasis and to evaluate the appearance of the patien's donor area, the recovery of transoral feeding function, and the presence of complications. Comparison of preoperative CDUS and CEUS findings and intraoperative exploration showed that CEUS had 100% sensitivity, specificity, positive predictive value, and negative predictive value for vascular exploration of the superficial inferior epigastric artery perforator flap, compared with 57%, 100%, 100%, and 25% for CDUS. The preoperative CDUS identified 25 penetrating vessels in 14 cases repaired with superficial inferior epigastric artery flaps. All vessel signals were enhanced by CEUS enhancement, and an additional 11 penetrating vessels were identified confirmed intraoperatively. The preoperative CEUS measurements of the initial diameter of superficial arteries in the abdominal wall were significantly higher than both CDUS and intraoperative measurements ( P<0.05); the difference in peak systolic velocity between CEUS and CDUS measurements was significant ( t=3.708, P=0.003). One case of superficial epigastric artery perforator flap developed venous embolism crisis at 48 hours after operation, the wound healing delayed. The other incisions in donor sites healed by first intention. All the patients were followed up 3-12 months, with an average of 8 months. No recurrence or metastasis appeared during the follow-up. There was no serious complications such as abdominal wall hernia, the location of abdominal scarring was hidden, and transoral feeding was resumed. The superficial inferior epigastric artery perforator flap with small injury in supply area and hidden scar location is a better choice for repairing oral and maxillofacial defects. The use of CEUS technique to assist the preoperative design of the superficial inferior epigastric artery perforator flap has good feasibility and high accuracy.

Influence of cuff-occlusion duration on contrast-enhanced ultrasound assessments of calf muscle microvascular blood flow responsiveness in older adults.

What is the central question of the study? Cuff-occlusion duration may influence contrast-enhanced ultrasound (CEUS) assessments of skeletal muscle microvascular blood flow responsiveness: what are the effects of 1, 3 and 5min cuff-occlusion on the magnitude and reliability of calf muscle microvascular blood flow responsiveness in older adults? What is the main finding and its importance? Calf muscle microvascular blood flow responsiveness was enhanced following 5min cuff-occlusion compared with 1min. The reliability of post-occlusion CEUS measurements was also improved following 5min occlusion. The use of a standardized 5min occlusion period should therefore be considered in future studies and clinical practice. Contrast-enhanced ultrasound (CEUS) is increasingly used in assessments of skeletal muscle microvascular blood flow responsiveness. In response to limb cuff-occlusion, some studies have reported significant impairments in CEUS measurements of microvascular blood flow in older adults with cardiovascular or metabolic disease, whereas others have failed to detect significant between-group differences, which has brought the reliability of the technique into question. In the absence of a standardized CEUS protocol, there is variance in the duration of cuff-occlusion used, which is likely to influence post-occlusion measurements of muscle microvascular blood flow. We aimed to determine the effect of cuff-occlusion duration by comparing the magnitude and reliability of CEUS measurements of calf muscle microvascular blood flow responsiveness in older adults (n=15, 67±11years) following 1, 3 and 5min occlusion periods. Microvascular blood flow (=microvascular volume × microvascular velocity) within the calf muscle was measured using real-time destruction-replenishment CEUS. Measurements were made following thigh cuff-occlusion (200mmHg) periods of 1, 3 and 5min in a random order. Microvascular blood flow was higher following 3min (3.71±1.46aUs-1 ) and 5min (3.47±1.48aUs-1 ) compared with 1min (2.42±1.27aUs-1 , P=0.002), which corresponded with higher microvascular volumes after 3 and 5min compared with 1min. Reliability was good following 5min (intraclass correlation coefficient (ICC) 0.49) compared with poor following 1 min (ICC 0.34) and 3 min (ICC 0.35). This study demonstrates that the magnitude and reliability of calf muscle microvascular responsiveness is enhanced using a 5min cuff-occlusion protocol compared with 1 min in older adults.

724 Contrast-Enhanced Ultrasound Measures of Skeletal Muscle Post-Occlusive Hyperaemia: What is the Effect of Occlusion Duration?

724 Contrast-Enhanced Ultrasound Measures of Skeletal Muscle Post-Occlusive Hyperaemia: What is the Effect of Occlusion Duration?

Perfusion controls muscle glucose uptake by altering the rate of glucose dispersion in vivo.

These studies test, using intravital microscopy (IVM), the hypotheses that perfusion effects on insulin-stimulated muscle glucose uptake (MGU) are 1) capillary recruitment independent and 2) mediated through the dispersion of glucose rather than insulin. For experiment 1, capillary perfusion was visualized before and after intravenous insulin. No capillary recruitment was observed. For experiment 2, mice were treated with vasoactive compounds (sodium nitroprusside, hyaluronidase, and lipopolysaccharide), and dispersion of fluorophores approximating insulin size (10-kDa dextran) and glucose (2-NBDG) was measured using IVM. Subsequently, insulin and 2[14C]deoxyglucose were injected and muscle phospho-2[14C]deoxyglucose (2[C14]DG) accumulation was used as an index of MGU. Flow velocity and 2-NBDG dispersion, but not perfused surface area or 10-kDa dextran dispersion, predicted phospho-2[14C]DG accumulation. For experiment 3, microspheres of the same size and number as are used for contrast-enhanced ultrasound (CEU) studies of capillary recruitment were visualized using IVM. Due to their low concentration, microspheres were present in only a small fraction of blood-perfused capillaries. Microsphere-perfused blood volume correlated to flow velocity. These findings suggest that 1) flow velocity rather than capillary recruitment controls microvascular contributions to MGU, 2) glucose dispersion is more predictive of MGU than dispersion of insulin-sized molecules, and 3) CEU measures regional flow velocity rather than capillary recruitment.

Contrast-enhanced ultrasound detects changes in microvascular blood flow in adults with sickle cell disease

In patients with sickle cell disease (SCD), poor outcome measures compromise the potential success of clinical trials. Contrast-enhanced ultrasound (CEUS) is a technique that can non-invasively quantify deep tissue microvascular blood flow. We tested the hypothesis that CEUS of forearm skeletal muscle could be used to: 1) assess microvascular abnormalities that occur during vaso-occlusive crisis; and 2) test new therapies for SCD that are targeted to improving the status of the microcirculation. We performed a prospective study, CEUS perfusion imaging of resting forearm muscle was performed in adults with SCD: 1) during and after a pain episode, and 2) before, during, and after a 24-hour infusion of the investigative agent, regadenoson, an adenosine A2A agonist. CEUS destruction-replenishment time-intensity data were analyzed to measure microvascular blood flow, as well as its components, microvascular blood volume and flux rate. Serial CEUS measurements were obtained in 32 adults with SCD. For the studies during crisis, there was a 30% reduction in microvascular blood flow compared to steady-state (p = 0.031), a reduction that was largely due to microvascular flux rate. For the regadenoson group, a non-significant 25% increase in flux rate and 9% increase in microvascular blood flow compared to baseline were detected during infusion. In a study of adults with SCD, CEUS detected changes in microvascular blood flow associated with vaso-occlusive crises. No changes were found during an infusion of the adenosine A2A agonist, regadenoson. This study provides preliminary evidence that CEUS could detect blood flow changes consistent with SCD physiology.

Comparison of dynamic contrast-enhanced magnetic resonance imaging and contrast-enhanced ultrasound for evaluation of the effects of sorafenib in a rat model of hepatocellular carcinoma

ObjectivesTo compare the accuracy of contrast-enhanced ultrasound (CEUS) and Dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) for the assessment of changes in tissue vascularization as result of sorafenib treatment in a rat model of hepatocellular carcinoma (HCC). MethodsMale Buffalo rats with orthotopic liver tumors treated daily with 7.5 mg/kg sorafenib via oral gavage for 2 weeks (n = 9) were subject to DCE-MRI and CEUS 2 weeks after tumor implantation - right before treatment initiation - and also after treatment completion - right before tumor harvest. Untreated animals (n = 10) were used as control. Tumor tissue sections were stained for hematoxylin-eosin, pimonidazole, and CD34 for quantitative assessment of necrosis, hypoxia, and microvessel density (MVD), respectively. ResultsOf all the DCE-MRI parameters that were evaluated, only volume transfer constant (Ktrans) measurements were significantly lower in sorafenib-treated tumors (0.18 vs 0.33 min−1, p < 0.01), indicating a substantial decrease in vascular permeability caused by the therapy. This reduction was associated with decreased MVD (3.9 vs 10.8% CD34+ cells, p < 0.01), higher tumor necrosis (31.9 vs 21.8%, p < 0.001) and hypoxia (19.7 vs 10.5% pimonidazole binding, p < 0.01). Moreover, statistical analysis demonstrate significant correlation of DCE-MRI Ktrans with histopathologic tissue necrosis (r = −0.537, p < 0.05) and MVD (r = 0.599, p < 0.05). Interestingly, none of the CEUS measurements were significantly different between the control and treatment groups, and did not show statistical correlation with any of the histopathological parameters assessed (p > 0.05). ConclusionsSorafenib-induced reduction in vascular permeability in this preclinical model of HCC is detected more accurately through DCE-MRI than CEUS, and DCE-MRI parameters strongly correlate with histopathological changes in tissue vascularization and tissue necrosis.

Contrast Enhanced Ultrasound with Submicron-Sized Contrast Agents Detects Diabetes Progression in Mouse Models of Type 1 Diabetes

In type 1 diabetes (T1D) the decline in beta-cell mass occurs many years prior to clinical presentation. A non-invasive method to diagnose insulitis and beta-cell mass decline prior to diabetes does not exist. During T1D progression, the islet microvasculature increases permeability due to insulitis. Iron oxide nanoparticle MRI contrast accumulation has previously been suggested as a measure of ongoing insulitis. Contrast enhanced ultrasound (CEUS) using gas-filled microbubbles (MBs) measures acoustic backscatter at sub-harmonic frequencies and is clinically approved. Sub-micron sized ‘nanobubbles’ (NBs) have previously been developed and show extravasation in tumors. Here we tested whether these sub-micron sized NBs show disease-mediated extravasation in mouse models of T1D. We performed CEUS measurements following NB infusion in NOD mice and control (NOD;Rag1ko and C57Bl6) mice. We also examined fluorescent coverage (FC) following rhodamine-labeled bubble delivery in these animals, through isolating pancreata for histology and quantifying FC and insulitis scores. We observed progressive increased CEUS signal accumulation in pancreas of 4w and 10w NOD mice, which was absent in NOD kidneys (p &amp;lt; 0.001). Rag1ko controls showed minimal CEUS signal in pancreas and kidney. The FC was limited to the pancreata of NOD mice and was substantially higher in endocrine compared to exocrine tissue (p = 0.002). NOD mouse islets also had substantially more FC compared to Rag1ko (p &amp;lt; 0.0001), and showed a correlation between NOD insulitis score and FC (p=0.043). Thus, CEUS with NB contrast agents can detect disease progression prior to T1D onset, which will be important for early T1D diagnosis and monitoring of disease prevention or reversal. Disclosure D. Ramirez: None. R.K. Benninger: None.

Contrast-enhanced ultrasound measurement of pancreatic blood flow dynamics predicts type 1 diabetes progression in preclinical models

In type 1 diabetes (T1D), immune-cell infiltration into the islets of Langerhans (insulitis) and β-cell decline occurs many years before diabetes clinically presents. Non-invasively detecting insulitis and β-cell decline would allow the diagnosis of eventual diabetes, and provide a means to monitor therapeutic intervention. However, there is a lack of validated clinical approaches for specifically and non-invasively imaging disease progression leading to T1D. Islets have a denser microvasculature that reorganizes during diabetes. Here we apply contrast-enhanced ultrasound measurements of pancreatic blood-flow dynamics to non-invasively and predictively assess disease progression in T1D pre-clinical models. STZ-treated mice, NOD mice, and adoptive-transfer mice demonstrate altered islet blood-flow dynamics prior to diabetes onset, consistent with islet microvasculature reorganization. These assessments predict both time to diabetes onset and future responders to antiCD4-mediated disease prevention. Thus contrast-enhanced ultrasound measurements of pancreas blood-flow dynamics may provide a clinically deployable predictive marker for disease progression in pre-symptomatic T1D and therapeutic reversal.

Quantitative evaluation of real-time maximum liver capacity (LiMAx) and time intensity curve (TIC) analysis in CEUS-based microperfusion.

To compare the diagnostic performance of real-time maximum liver capacity (LiMAx) with dynamic contrast-enhanced ultrasound (CEUS)-based liver microcirculation. 23 patients underwent liver function capacity (LiMAx) test and consecutive or previous CEUS examinations. A bolus injection of 1.4 ml sulfur hexafluoride microbubbles was administered for CEUS measurements (1-6 MHz) and quantitative perfusion analysis (TIC) was performed with an integrated perfusion software using stored cine-loops. Two perfusion-parameters, time to peak (TtoP) and area under the curve (Area), were evaluated in liver parenchyma and portal vein using TIC analysis.To compare quantification parameters, patients were classified in patients representing a healthy population (LiMAx value >315 μg/kg/h) and those representing patients with liver disease (LiMAx value <315 μg/kg/h). Comparing perfusion parameters derived from portal vein measurements, TtoP and Area were higher in patients with normal liver function TtoP: 25.0±8.4 s, Area: 1483±920 a.u. compared to patients with impaired liver function TtoP: 22.4±14.0 s; Area 1351±1212 a.u. This difference however was not statistically significant (p = 0.52, p = 0.48).In parameters derived from measurements in liver parenchyma TtoP was higher (38.5±11.3 s) and Area was lower (999±632 a.u.) in patients with normal liver function compared to patients with impaired liver function (TtoP; 30.6±11.0 s, p = 0.156; Area: 1202±719 a.u.) (p = 0.16, p = 0.56).In a simple linear regression model, none of the perfusion parameters measured in portal vein (TtoP portal, Area portal) and liver parenchyma (TtoP liver, Area liver) correlated significantly with respective LiMAx values (p = 0.194-0.950). Within the framework of this study, CEUS-based perfusion parameters were not able to assess severity of liver disease, assessed with LiMAx- test.