Microbubbles and ultrasound: from diagnosis to therapy.

The development of ultrasound contrast agents, containing encapsulated microbubbles, has increased the possibilities for diagnostic imaging. Ultrasound contrast agents are currently used to enhance left ventricular opacification, increase Doppler signal intensity, and in myocardial perfusion imaging. Diagnostic imaging with contrast agents is performed with low acoustic pressure using non-linear reflection of ultrasound waves by microbubbles. Ultrasound causes bubble destruction, which lowers the threshold for cavitation, resulting in microstreaming and increased permeability of cell membranes. Interestingly, this mechanism can be used for delivery of drugs or genes into tissue. Microbubbles have been shown to be capable of carrying drugs and genes, and destruction of the bubbles will result in local release of their contents. Recent studies demonstrated the potential of microbubbles and ultrasound in thrombolysis. In this article, we will review the recent advances of microbubbles as a vehicle for delivery of drugs and genes, and discuss possible therapeutic applications in thrombolysis.     

Targeted ultrasound imaging using microbubbles

Targeted ultrasound imaging uses acoustically active contrast agents bearing a ligand on the surface that binds to a function-specific molecule. These ultrasound contrast agents are typically gas-filled microbubbles, nongaseous liposomes, or lipid-encapsulated perfluorocarbon emulsions. Binding of the contrast agent to the target results in persistent contrast enhancement during ultrasound imaging. This approach has been applied to the ultrasound imaging of pathophysiologic processes such as inflammation associated with ischemia reperfusion, heart transplant rejection, atherosclerotic plaque, thrombus, and apoptosis.     

Thinking outside the "box"-the ultrasound contrast controversy

On October 10, 2007, the U.S. Food and Drug Administration (FDA) announced a new "black box" warning for the perflutren-containing ultrasound contrast agents, contraindicating their use in patients with acute coronary syndromes, acute myocardial infarction, and worsening or clinically unstable heart failure. These warnings ignore the proven efficacy of ultrasound contrast agents, the previously established safety of these compounds, the potential risks of alternative procedures, and the likely confounding effect of pseudocomplication. We suggest that the FDA Medical Imaging Division convene a panel of cardiologists experienced in a variety of imaging modalities to fully assess the adverse events that have been attributed to these agents and that any future FDA warnings acknowledge the possible influence of pseudocomplication, the proven efficacy of the modality in question for early and accurate diagnosis of cardiovascular disease, and the known and theoretical risks of alternative testing that may be necessary.     

Contrast echocardiography

Myocardial contrast echocardiography (MCE) is a noninvasive imaging technique that relies on the ultrasound detection of microbubble contrast agents. These agents are confined to the intravascular space thereby producing signal enhancement from the blood pool. This review encompasses many of the key concepts regarding the clinical application of MCE. The first section focuses on the composition, safety, and biokinetics of ultrasound contrast agents. Then we discuss new ultrasound imaging methodology that has been developed to enhance detection of contrast agent and to assess perfusion at the tissue level. Next, the clinical applications of contrast ultrasound are reviewed. These include enhancement of the cardiac chambers for better assessment of cardiac function and masses, myocardial perfusion imaging for the detection of coronary artery disease, and the assessment of myocardial viability and microvascular reflow. Finally, we discuss some of the future applications for MCE, which include molecular imaging of disease and drug/gene delivery. The overall aim of the review is to update the clinician on state-of-the-art MCE and how it can be applied in patients with cardiovascular disease.     

Contrast‐enhanced ultrasound for quantification of tissue perfusion in humans

Contrast‐enhanced ultrasound is an imaging technique that can be used to quantify microvascular blood volume and blood flow of vital organs in humans. It relies on the use of microbubble contrast agents and ultrasound‐based imaging of microbubbles. Over the past decades, both ultrasound contrast agents and experimental techniques to image them have rapidly improved, as did experience among investigators and clinicians. However, these improvements have not yet resulted in uniform guidelines for CEUS when it comes to quantification of tissue perfusion in humans, preventing its uniform and widespread use in research settings. The objective of this review is to provide a methodological overview of CEUS and its development, the influences of hardware and software settings, type and dosage of ultrasound contrast agent, and method of analysis on CEUS‐derived perfusion data. Furthermore, we will discuss organ‐specific imaging challenges, advantages, and limitations of CEUS.     

Ultrasound molecular imaging of cardiovascular disease

Myocardial contrast echocardiography utilizes intravenously injected gas-filled microspheres as acoustically active red blood cell tracers. During ultrasound imaging, unimpeded microsphere transit through the intramyocardial microcirculation causes transient myocardial opacification, which can be mapped and quantified as myocardial perfusion. Ultrasound molecular imaging utilizes similar acoustically active microspheres, which are modified to bear a receptor-specific ligand on the surface, conferring microsphere binding to a disease-specific endothelial epitope. Because the microspheres adhere to the endothelium, ultrasound imaging reveals a persistent, rather than transient, contrast effect, indicating the presence and location of the molecule of interest in real time. Molecular contrast echocardiography has been developed to detect upregulated leukocyte adhesion molecules during microvascular inflammation, such as occurs in cardiac transplant rejection and ischemia-reperfusion. Principles of microsphere targeting and ultrasound imaging of microvascular epitopes have been extended to larger vessels to image molecular markers of atherosclerosis. This Article summarizes the current status of cardiovascular ultrasound molecular imaging. Experimental proofs of concept will be outlined and the clinical extension of these concepts to the molecular imaging of cardiovascular disease using clinical ultrasound technology will be discussed.     

Improved myocardial contrast with second harmonic transient ultrasound response imaging in humans using intravenous perfluorocarbon-exposed sonicated dextrose albumin

Objectives. The objectives of this study were to determine whether a new method of ultrasound imaging (transient response imaging) could improve the myocardial contrast after intravenous injections of perfluorocarbon-exposed sonicated dextrose albumin microbubble contrast medium in humans.Background. We have shown in animals that very low doses of intravenous contrast medium can produce transient but significantly better myocardial contrast when diagnostic ultrasound pulses are interrupted (delivered only once per cardiac cycle) instead of conventional 25- to 30-Hz frame rate imaging.Methods. In 14 patients with normal rest wall motion, the peak myocardial contrast produced by transient response imaging was compared with that produced by conventional harmonic ultrasound imaging after injections of low doses (0.0025 to 0.01 ml/kg) of intravenous contrast medium. All studies were performed with second harmonic imaging (2.0 to 2.5 MHz-transmitted frequency). Blood pressure, oxygen saturation, respiratory rate and pulse were monitored before and after each injection.Results. The intravenous contrast medium in the doses given produced no hemodynamic changes and no significant side effects in any patients. Overall, the mean (± SD) anterior and posterior myocardial contrast produced was significantly greater with transient response imaging than with conventional harmonic ultrasound imaging (anterior: 37 ± 20 U transient response imaging vs. 18 ± 14 U conventional harmonic imaging, posterior: 17 ± 14 U transient response imaging vs. 5 ± 5 U conventional; p < 0.01). With the sample size of 14 patients, the study had 80% power to detect a tree difference of 18 U for anterior myocardial contrast and 90% power to detect a difference of 12 U for posterior contrast. Visually evident anterior or apical myocardial contrast was observed in 14 of 15 patients with transient response imaging but in only 7 patients with conventional harmonic imaging. Posterior or basal myocardial contrast was evident in 10 patients with transient response imaging but in only 1 patient with conventional harmonic imaging.Conclusions. Transient response imaging produces significantly better myocardial contrast than conventional harmonic imaging in humans and can be produced safely with minute quantities of intravenous perfluorocarbon.     

Ultrasound Contrast Materials in Cardiovascular Medicine: from Perfusion Assessment to Molecular Imaging

Ultrasound imaging is widely used in cardiovascular diagnostics. Contrast agents expand the range of tasks that ultrasound can perform. In the clinic in the USA, endocardial border delineation and left ventricle opacification have been an approved indication for more than a decade. However, myocardial perfusion contrast ultrasound studies are still at the clinical trials stage. Blood pool contrast and perfusion in other tissues might be an easier indication to achieve: general blood pool ultrasound contrast is in wider use in Europe, Canada, Japan, and China. Targeted (molecular) contrast microbubbles will be the next generation of ultrasound imaging probes, capable of specific delineation of the areas of disease by adherence to molecular targets. The shell of targeted microbubbles (currently in the preclinical research and early stage clinical trials) is decorated with the ligands (antibodies, peptides or mimetics, hormones, and carbohydrates) that ensure firm binding to the molecular markers of disease.     

Detection of Myocardial Perfusion in Multiple Echocardiographic Windows With One Intravenous Injection of Microbubbles Using Transient Response Second Harmonic Imaging

Objectives. The purpose of this study was to prove that transient response harmonic imaging could detect normal and abnormal myocardial perfusion in multiple echocardiographic windows with one intravenous injection of microbubbles in humans.Background. Myocardial ultrasound contrast can be produced from intravenous perfluorocarbon-exposed sonicated dextrose albumin, and ultrasound can be significantly improved by briefly suspending the interval between frame rates. Whether this contrast can noninvasively quantify myocardial perfusion in humans is unknown.Methods. In 28 patients, harmonic transient response imaging was used to image the heart in multiple different imaging planes after one intravenous injection of ultrasound contrast agent. Twenty-five of these 28 patients had a repeat injection during dipyridamole stress. In the primary view, the ultrasound transmission rate was one frame per cardiac cycle; in secondary and tertiary views, the transmission rate was once every multiple cardiac cycles. Regional myocardial contrast was visually assessed and quantified off-line. Quantitative rest thallium and dipyridamole stress sestamibi imaging was also performed.Results. Perfusion abnormalities were evident in the secondary and tertiary views only with one frame every multiple cardiac cycles. Regional peak myocardial videointensity (PMVI) correlated closely with regional tracer uptake in individual patients both at rest (r = 0.84) and during stress (r = 0.88). A PMVI ratio (abnormal region divided by the region with highest nuclear uptake) <0.6 in any view had a 92% sensitivity and a 84% specificity in identifying a regional nuclear perfusion abnormality.Conclusions. Transient response imaging produces myocardial contrast in multiple views with one intravenous injection of contrast agent and can accurately identify regional myocardial perfusion abnormalities.(J Am Coll Cardiol 1997;29:791–9)     

Disappearing liver metastases from colorectal cancer: impact of modern imaging modalities

BackgroundChemotherapy is often used before a resection for colorectal liver metastases. After chemotherapy, metastases may disappear on cross sectional imaging but residual metastatic disease may still exist. The aim of this retrospective study was to investigate the impact of new advancements in imaging technology such as magnetic resonance imaging (MRI) with liver specific contrast (Gd EOB DTPA) and contrast enhanced intra operative ultrasound (CE IOUS) on disappearing liver metastases (DLM).MethodsTwenty nine patients with one or more DLM undergoing surgical exploration were included. Pre operative imaging consisted of contrast enhanced multi detector computed tomography (MDCT) and/or MRI with liver specific contrast. At surgery, CE IOUS was used when tumours known from pre chemotherapy imaging were not found by inspection or intra operative ultrasound.ResultsPatients presented 66 DLM. At surgical exploration, 42 DLM were identified and treated (64%). CE IOUS detected one additional DLM not found by intra operative ultrasound. For metastases ≤10 mm on histological analysis, imaging sensitivities for MRI and MDCT before surgery but after chemotherapy were 26/49 (53%) and 24/66 (36%), respectively.ConclusionA majority of DLM are identified during surgery using intra operative ultrasound, with only little additional value of CE IOUS. The sensitivities of post chemotherapy imaging modalities for small metastases are low in the setting of DLM. For surgical planning, an optimized pre chemotherapy imaging is essential.     

The ultrasound contrast imaging properties of lipid microbubbles loaded with urokinase in dog livers and their thrombolytic effects when combined with low-frequency ultrasound in vitro

A new microbubble loaded with urokinase (uPA-MB) was explored in a previous study. However, its zeta potential and ultrasound contrast imaging properties and its thrombolytic effects when combined with low-frequency ultrasound (LFUS) were unclear. The zeta potential and ultrasound contrast imaging property of 5 uPA-MBs loading with 50,000 IU uPA was respectively detected using a Malvern laser particle analyzer and a Logiq 9 digital premium ultrasound system. Its ultrasound contrast imaging property was performed on the livers of two healthy dogs to compare with SonoVue. And the clot mass loss rate, D-dimer concentration and surface morphology of the clot residues were measured to evaluate the thrombolytic effect after treatment with three doses of 5 uPA-MBs combined with LFUS in vitro. The zeta potential of 5 uPA-MBs (?27.0 ± 2.40 mV) was higher than that of normal microbubbles (?36.95 ± 1.77 mV). Contrast-enhanced imaging of the hepatic vessels using 5 uPA-MBs was similar to SonoVue, while the imaging duration of 5 uPA-MBs (10 min) was longer than SonoVue (6 min). The thrombolytic effect of three doses of uPA-MBs combined with LFUS was significantly better than that of the control group and showed dose dependence. The 5 uPA-MBs have a negative charge on their surface and good echogenicity as ultrasound contrast agents. The 5 uPA-MBs combined with LFUS can promote thrombolysis in a dose-dependent manner.     

Contrast echocardiography: An update

Despite the advent of tissue harmonic imaging, echocardiography fails to produce diagnostically useful images in a significant proportion of patients. This often leads to inaccurate assessment of left ventricular function, necessitating the use of other, more expensive and laborious imaging techniques, purely for diagnostic purposes. This has facilitated the development of microbubbles, which together with ultrasound, produce opacification of the left ventricular cavity, thus enabling clear visualization and accurate quantification of left ventricular function. Contrast agents have also facilitated the development of myocardial contrast echocardiography. This allows assessment of cardiac anatomy, function, and perfusion, all in one sitting, by the bedside. Contrast ultrasound imaging also has now been applied to newer techniques (eg, real-time three-dimensional echocardiography) and is also showing promise in other cardiovascular scans (eg, carotid ultrasound for intima-media thickness). Thus, contrast agents play a pivotal role in noninvasive cardiovascular imaging and its use worldwide is likely to increase.     

Real-Time Contrast Echo Assessment of Myocardial Perfusion at Low Emission Power: First Experimental and Clinical Results Using Power Pulse Inversion Imaging

Power pulse inversion (PPI) has been developed for echocontrast specific imaging in order to reduce destruction of microbubbles. The purpose of this study was to evaluate PPI for real-time contrast echocardiography. Therefore, in vitro studies in a physiological flow-phantom and clinical examinations in patients with coronary artery disease were performed. The in vitro rersults of this study indicate that PPI allows real-time imaging at low emission power and is almost nondestructive to contrast microbubbles of Definity. At this low emission power a strong linear relationship between the dosage of the contrast agent and the resulting PPI signal intensity was found (R = 0.998, p < 0.001). In the clinical examinations real-time imaging using low mechanical index PPI resulted in strong myocardial signals and a complete filling of the cavities indicating absence of bubble destruction. Most striking was the ability of PPI to display myocardial thickening and wall motion simultaneously with the assessment of myocardial contrast replenishment following ultrasound induced bubble destruction by high power frames. We conclude that PPI allows nondestructive contrast imaging both in experimental and clinical settings. Therefore, real-time imaging of myocardial perfusion and real-time assessment of contrast replenishment following ultrasound induced destruction of microbubbles is feasible. Moreover, PPI allows simultaneous assessment of perfusion and myocardial function.     

Molecular ultrasound assessment of tumor angiogenesis

Angiogenesis, the growth of new blood vessels, plays a critical role in progression of tumor growth and metastasis, making it an attractive target for both cancer imaging and therapy. Several molecular markers, including those that are involved in the angiogenesis signaling pathway and those unique to tumor angiogenic vessels, have been identified and can be used as targets for molecular imaging of cancer. With the introduction of ultrasound contrast agents that can be targeted to those molecular markers, targeted contrast-enhanced ultrasound (molecular ultrasound) imaging has become an attractive imaging modality to non-invasively assess tumor angiogenesis at the molecular level. The advantages of molecular ultrasound imaging such as high temporal and spatial resolution, non-invasiveness, real-time imaging, relatively low cost, lack of ionizing irradiation and wide availability among the imaging community will further expand its roles in cancer imaging and drug development both in preclinical research and future clinical applications.     

Contrast echocardiography for myocardial perfusion imaging using intravenous agents: progress and promises.

AIMS: This article is a convenient overview to assist the interested echocardiographist towards acquiring his own experience in the field of myocardial perfusion imaging using intravenous contrast agents. This goal is now pursued in many centres, since contrast echo holds the advantages of cardiac ultrasound (non-invasiveness, high spatial and temporal resolution, wide availability, use of non-ionizing radiation), and because a variety of transpulmonary agents-together with a spectrum of imaging modalities-are becoming available. METHODS AND RESULTS: Many technical considerations need to be addressed for optimal myocardial perfusion imaging: characteristics of the contrast medium (air-filled or perfluorocarbon filled and/or encapsulated agents), modality of administration (bolus injection or continuous infusion) and interaction between microbubbles and ultrasound (dependency on power output). Moreover, intermittent harmonic imaging, intermittent harmonic power Doppler, pulse inversion and amplitude modulation imaging have all been developed to enhance microbubble detection over myocardial tissue. These new acquisition modalities also yield specific artifacts impacting on myocardial perfusion assessment. Finally, acute myocardial infarction and chronic ischaemic heart disease (at baseline and during stress) are the most studied clinical models for perfusion imaging with contrast echo, and are reviewed in this article. CONCLUSION: Perfusion imaging with intravenous contrast agents has never been as close to widespread clinical use as it is today, but many methodological issues remain unsettled before the wish of the contrast echocardiographist comes true: that is, a cheap, user-friendly and widely available technology that would disclose new information in echocardiography.     

Functionalized multiwalled carbon nanotubes as ultrasound contrast agents

Ultrasonography is a fundamental diagnostic imaging tool in everyday clinical practice. Here, we are unique in describing the use of functionalized multiwalled carbon nanotubes (MWCNTs) as hyperechogenic material, suggesting their potential application as ultrasound contrast agents. Initially, we carried out a thorough investigation to assess the echogenic property of the nanotubes in vitro. We demonstrated their long-lasting ultrasound contrast properties. We also showed that ultrasound signal of functionalized MWCNTs is higher than graphene oxide, pristine MWCNTs, and functionalized single-walled CNTs. Qualitatively, the ultrasound signal of CNTs was equal to that of sulfur hexafluoride (SonoVue), a commercially available contrast agent. Then, we found that MWCNTs were highly echogenic in liver and heart through ex vivo experiments using pig as an animal model. In contrast to the majority of ultrasound contrast agents, we observed in a phantom bladder that the tubes can be visualized within a wide variety of frequencies (i.e., 5.5–10 MHz) and 12.5 MHz using tissue harmonic imaging modality. Finally, we demonstrated in vivo in the pig bladder that MWCNTs can be observed at low frequencies, which are appropriate for abdominal organs. Importantly, we did not report any toxicity of CNTs after 7 d from the injection by animal autopsy, organ histology and immunostaining, blood count, and chemical profile. Our results reveal the enormous potential of CNTs as ultrasound contrast agents, giving support for their future applications as theranostic nanoparticles, combining diagnostic and therapeutic modalities.     

Contrast-enhanced endoscopic ultrasound with low mechanical index: a new technique

AIM: Recent advances in technology have supported the development of new endoscopic ultrasound systems making it possible to use low MI contrast-enhanced imaging techniques (wide band harmonic imaging done with endoscopic ultrasound is currently at a preliminary stage). We now report on the first use of contrast-enhanced, low mechanical index, real-time endoscopic ultrasound (CELMI-EUS) in six patients using prototype technology. MATERIALS AND METHODS: CELMI-EUS was performed using an electronic echo-endoscope HITACHI/Pentax EG-3830UT and adapted dynamic contrast harmonic wide-band pulsed inversion software with low mechanical index (MI = 0.09 - 0.25) before and up to 180 seconds after injection of SonoVue (4.8 mL) in six patients. RESULTS: Adequate visualisation of the arterial and portal venous phases was achieved in all patients. The pancreas and liver were studied thereafter. In contrast to the satisfactory visualisation of these vessels, enhancement of the left liver lobe was sufficient only in 4 patients. In the remaining 2 patients with liver cirrhosis, the enhancement was less pronounced in contrast to the strong enhancement of the hepatic artery and portal vein. CONCLUSION: Recent advances in technology have supported the development of new echo-endoscopic systems making it possible to use real-time, low mechanical index, contrast-enhanced imaging techniques with endoscopic ultrasound. We have preliminarily shown that arterial, portal venous and parenchymal contrast enhancement is possible.     

One stop shop approach for the diagnosis of liver hemangioma

Hepatic hemangioma is usually detected on a routine ultrasound examination because of silent clinical behaviour. The typical ultrasound appearance of hemangioma is easily recognizable and quickly guides the diagnosis without the need for further investigation. But there is also an entire spectrum of atypical and uncommon ultrasound features and our review comes to detail these particular aspects. An atypical aspect in standard ultrasound leads to the continuation of explorations with an imaging investigation with contrast substance [ultrasound/ computed tomography/or magnetic resonance imaging (MRI)]. For a clinician who practices ultrasound and has an ultrasound system in the room, the easiest, fastest, non-invasive and cost-effective method is contrast enhanced ultrasound (CEUS). Approximately 85% of patients are correctly diagnosed with this method and the patient has the correct diagnosis in about 30 min without fear of malignancy and without waiting for a computer tomography (CT)/MRI appointment. In less than 15% of patients CEUS does not provide a conclusive appearance; thus, CT scan or MRI becomes mandatory and liver biopsy is rarely required. The aim of this updated review is to synthesize the typical and atypical ultrasound aspects of hepatic hemangioma in the adult patient and to propose a fast, non-invasive and cost-effective clinical-ultrasound algorithm for the diagnosis of hepatic hemangioma.     

Ultrasound of acute abdomen

Ultrasound is an important tool in the diagnosis of acute abdominal pain, which is mainly caused by gastrointestinal diseases. This article gives an overview on the important differential diagnosis related to pain localization. Sonomorphological signs and their sensitivity and specificity are discussed. In contrast to other imaging methods ultrasound is hand guided. The present article differentiates the diagnostic capability of ultrasonic diagnosis depending on technical equipment and sonographer's educational level. These facts are important to stabilize the position of ultrasound in the ensemble of other imaging methods. The use of mobile ultrasound machines and an improved training (e.g. by computer assisted sonosimulator systems) will lead to an increasing importance of ultrasound.     

Grundlagen der Sonographie und deren Relevanz für die Innere Medizin

This article gives an overview of the technical possibilities and clinical applications of modern ultrasound. B-mode and Doppler imaging are already essential in daily routine. Advanced technology, such as harmonic imaging, improves image quality. Tissue harmonic imaging provides better resolution by reducing noise, whereas the introduction of contrast agents increases the perceptibility of focal lesions, e.g., liver tumors. Meanwhile the diagnostic accuracy of contrast-enhanced ultrasound imaging is comparable with CT imaging. Other tools, e.g., elastography, ameliorate the detection of liver fibrosis and tumors. Currently, application of three-dimensional and four-dimensional imaging is of low clinical relevance. Panoramic imaging provides images close to CT or MRI.