Mechanisms of microbubble-vessel interactions and induced stresses: a numerical study.

Oscillating microbubbles within microvessels could induce stresses that lead to bioeffects or vascular damage. Previous work has attributed vascular damage to the vessel expansion or bubble jet. However, ultra-high speed images of recent studies suggest that it could happen due to the vascular invagination. Numerical simulations of confined bubbles could provide insight into understanding the mechanism behind bubble-vessel interactions. In this study, a finite element model of a coupled bubble/fluid/vessel system was developed and validated with experimental data. Also, for a more realistic study viscoelastic properties of microvessels were assessed and incorporated into this comprehensive numerical model. The wall shear stress (WSS) and circumferential stress (CS), metrics of vascular damage, were calculated from these simulations. Resultant amplitudes of oscillation were within 15% of those measured in experiments (four cases). Among the experimental cases, it was numerically found that maximum WSS values were between 1.1-18.3 kPa during bubble expansion and 1.5-74 kPa during bubble collapse. CS was between 0.43-2.2 MPa during expansion and 0.44-6 MPa while invaginated. This finding confirmed that vascular damage could occur during vascular invaginations. Predicted thresholds in which these stresses are higher during vessel invagination were calculated from simulations.

The Characteristics of Vascular Growth in VX2 Tumor Measured by MRI and Micro-CT.

Blood supply is crucial for rapid growth of a malignant tumor; medical imaging can play an important role in evaluating the vascular characterstics of tumors. Magnetic resonance imaging (MRI) and micro-computed tomography (CT) are able to detect tumors and measure blood volumes of microcirculation in tissue. In this study, we used MR imaging and micro-CT to assess the microcirculation in a VX2 tumor model in rabbits. MRI characterization was performed using the intravascular contrast agent Clariscan (NC100150-Injection); micro-CT with Microfil was used to directly depict blood vessels with diameters as low as 17 um in tissue. Relative blood volume fraction (rBVF) in the tumor rim and blood vessel density (rBVD) over the whole tumor was calculated using the two imaging methods. Our study indicates that rBVF is negatively related to the volume of the tumor measured by ultrasound (R = 0.90). rBVF in the tissue of a VX2 tumor measured by MRI in vivo was qualitatively consistent with the rBVD demonstrated by micro-CT in vitro (R = 0.97). The good correlation between the two methods indicates that MRI studies are potentially valuable for assessing characteristics or tumor vascularity and for assessing response to therapy noninvasively.

Hepatic perfusion imaging using factor analysis of contrast enhanced ultrasound.

Contrast enhanced ultrasound imaging provides a real-time tool for evaluating vasculature in the liver. Primary liver cancer is known to be perfused exclusively by blood from the hepatic artery, whereas normal liver is also supplied by the portal vein. Visual separation of two different phases of enhancement from the independent feeding vessels is important for diagnosis but remains a challenge. This paper presents a method of using factor analysis for extracting distinct time-intensity curves. A key component to this extraction is the clustering of measured bolus curves and their projection onto a positivity domain to obtain nonnegative curves. This technique provides complementary images representing spatial loadings on each curve. As little as 1% of the data is required to contain unmixed signals to extract time-intensity curves that correlate well with true curves. A method of combining this information to display a regional hepatic perfusion image is proposed, and results are tested on a set of 10 patients. Region of interest analysis suggests it is possible to detect changes in the hepatic perfusion index of liver lesions relative to normal liver parenchyma using contrast ultrasound.

High-frequency subharmonic pulsed-wave Doppler and color flow imaging of microbubble contrast agents.

A recent study has shown the feasibility of subharmonic (SH) flow imaging at a transmit frequency of 20 MHz. This paper builds on these results by examining the performance of SH flow imaging as a function of transmit pressure. Further, we also investigate the feasibility of SH pulsed-wave Doppler (PWD) imaging. In vitro flow experiments were performed with a 1-mm-diameter wall-less vessel cryogel phantom using the ultrasound contrast agent Definity and an imaging frequency of 20 MHz. The phantom results show that there is an identifiable pressure range where accurate flow velocity and power estimates can be made with SH imaging at 10 MHz (SH10), above which velocity estimates are biased by radiation force effects and unstable bubble behavior, and below which velocity and power estimates are degraded by poor SNR. In vivo validation of SH PWD was performed in an arteriole of a rabbit ear, and blood velocity estimates compared well with fundamental (F20) mode PWD. The ability to suppress tissue signals using SH signals may enable the use of higher frame rates and improve sensitivity to microvascular flow or slow velocities near large vessel walls by reducing or eliminating the need for clutter filters.

In vitro characterization of the subharmonic ultrasound signal from Definity microbubbles at high frequencies.

Ultrasound microbubble contrast agents have been demonstrated to scatter subharmonic energy at one-half the driving frequency. At ultrasound frequencies in the 20-40 MHz range, the subharmonic offers the potential to differentiate the blood in the microcirculation from the surrounding tissue. It is unknown whether current contrast agents, manufactured to be resonant between 2 and 12 MHz, are ideal for subharmonic imaging at higher frequencies. We performed numerical simulations of the Keller-Miksis model for the behavior of a single bubble and experimental investigations of Definity microbubbles in water. The results supported the hypothesis that off-resonant bubbles, excited at their second harmonic, may be primarily responsible for the observed subharmonic energy. For frequencies between 20 and 32 MHz and 32 and 40 MHz, the optimal bubble diameters for the generation of subharmonics in vitro were determined experimentally to be 1.2-5 microm and less than 1.2 microm, respectively. Definity may be a suitable ultrasound contrast agent for subharmonic imaging at 20 MHz with peak-negative pressures between 380 and 590 kPa and pulses greater than or equal to four cycles in duration.

Liver mass evaluation with ultrasound: the impact of microbubble contrast agents and pulse inversion imaging.

Liver mass evaluation includes two essential elements--lesion detection and lesion characterization. Both of these are greatly improved on sonography with the addition of contrast agents and the use of specialized imaging techniques, particularly pulse inversion imaging. Ultrasound contrast agents are comprised of tiny microbubbles of gas that interact with the ultrasound beam producing an enhancement of the Doppler signal from blood. Pulse inversion imaging allows preferential detection of the signal from the microbubble agents with suppression of the signal from background tissue. Two imaging techniques include a low mechanical index (MI) nondestructive method to show lesional vascularity and a high MI destructive mode that produces disruption of the bubbles in a single frame. The latter allows for quantitative assessment of the relative enhancement of a lesion as compared with the adjacent liver parenchyma, which is a reflection of the relative vascular volumes. Vascular imaging has shown characteristic and reproducible features of common liver masses, including hemangioma, focal nodular hyperplasia, hepatocellular carcinoma, and liver metastases. Delayed postvascular enhancement of the normal liver, a phenomenon that is unique to certain classes of microbubble contrast agents, allows detection of more and smaller malignant lesions than on baseline.

Tissue harmonic imaging: is it a benefit for bile duct sonography?

OBJECTIVE: Our purpose was to compare tissue harmonic imaging with conventional sonography of the biliary tract. SUBJECTS AND METHODS: Eighty patients with suspect biliary disease had conventional sonography and tissue harmonic imaging with an ATL 3000 or 5000 scanner in a 6-month interval. Final diagnoses included malignant biliary obstruction (n = 30), choledocholithiasis (n = 16), sclerosing cholangitis (n = 4), normal or nonobstructed ducts (n = 16), and miscellaneous conditions (n = 14). Similar images were taken with each technique in terms of projection, field of view, focal zone selection, and evidence of disease. Two separate observers blinded to patient data and technique reviewed and graded images individually for the appearance of the lumen of the bile ducts, the length of the visible duct, the appearance of the duct wall, the presence of any intraluminal masses, and the appearance of associated acoustic shadows. Images were graded from zero to 3, with 3 being the best. RESULTS: The median of the 546 tissue harmonic images was one grade higher than the median for the corresponding conventional images (p < 0.0001). Improvements with tissue harmonic imaging included better sharpness of the duct walls (p < 0.01), a clearer lumen (p < 0.0001), identification of a longer length of the common bile duct (p < 0.0001), and improved detection of intraluminal masses (p < 0.006). Acoustic shadows were better defined and blacker with tissue harmonic imaging (p < 0.007). CONCLUSION: Improvement in contrast and reduction of side lobe artifacts with tissue harmonic imaging enhance visualization of the biliary ducts. Tissue harmonic imaging is now our routine technique for bile duct examination.

Techniques for perfusion imaging with microbubble contrast agents.

The acoustic properties of ultrasound contrast agents vary widely with agent composition and insonation conditions. For contrast imaging, methods are required to match RF and Doppler processing to each combination of transmission parameters and agent and tissue properties. We propose a method that uses the measured or modeled echoes from agent and tissue to specify directly the characteristics of RF and Doppler filters for contrast imaging. The proposed method is sufficiently general to cover most common imaging techniques including harmonic greyscale, Doppler, and pulse inversion imaging. Using this method, sample filters were designed to detect myocardial perfusion with the contrast agent Optison (Mallinckrodt Medical, St. Louis, MO) under selected imaging conditions. Simplified power Doppler filtering, using a weighted sum of the Doppler samples, matched the performance of more complicated matrix filters. By coordinating the selection of RF and Doppler filters rather than designing these filters sequentially, agent-to-tissue contrast was increased by up to 3.9 dB. Under some conditions, fundamental RF filtering outperformed harmonic filtering for intermittent Doppler imaging.

Predicting the acoustic response of a microbubble population for contrast imaging in medical ultrasound.

Although the behavior of a bubble in an acoustic field has been studied extensively, few theoretical treatments to date have been applied to simulate the acoustic response of a real population of variably sized microbubbles in a finite-width sound beam. In this paper, we present a modified Trilling equation for single bubble dynamics that has been solved numerically for different conditions. Radiated waveforms from a large number of such bubbles are combined, reflecting their size distribution and location and the shape of a real acoustic beam. The resulting time-domain pressure waveforms can be compared with those obtained experimentally. The dependence of second-harmonic radiation on incident focal amplitude at different frequencies is presented. This model is particularly suited to the study of interaction between a medical ultrasound beam and microbubble contrast agents in aqueous media.

Contrast echocardiography: current and future applications.

Recent updates in the field of echocardiography have resulted in improvements in image quality, especially in those patients whose ultrasonographic (ultrasound) evaluation was previously suboptimal. Intravenous contrast agents are now available in the United States and Europe for the indication of left ventricular opacification and enhanced endocardial border delineation. The use of contrast enables acquisition of ultrasound images of improved quality. The technique is especially useful in obese patients and those with lung disease. Patients in these categories comprise approximately 10% to 20% of routine echocardiographic examinations. Stress echocardiography examinations can be even more challenging, as the image acquisition time factor is critically important for accurate detection of coronary disease. Improvements in image quality with intravenous contrast agents can facilitate image acquisition and enhance delineation of regional wall motion abnormalities at the peak level of exercise. Recent phase III clinical trial data on the use of Optison and several other agents (currently under evaluation) have revealed that for approximately half of patients, image quality substantively improves, which enables the examination to be salvaged and/or increases diagnostic accuracy. For the "difficult-to-image" patient, this added information results in (1) enhanced laboratory efficiency, (2) a reduction in downstream testing, and (3) possible improvements in patient outcome. In addition, substantial research efforts are underway to use ultrasound contrast agents for assessment of myocardial perfusion. The detection of myocardial perfusion during echocardiographic examinations will permit the simultaneous assessment of global and regional myocardial structure, function, and perfusion-all of the indicators necessary to enable the optimal noninvasive assessment of coronary artery disease. Despite the added benefit in improved efficacy of testing, few data exist regarding the long-term effectiveness of these agents. Currently under evaluation are the clinical and economic outcome implications of intravenous contrast agent use for daily clinical decision making in a variety of patient subsets. Until these data are known, this document offers a preliminary synthesis of available evidence on the value of intravenous contrast agents for use in rest and stress echocardiography. At present, it is the position of this guideline committee that intravenous contrast agents demonstrate substantial value in the difficult-to-image patient with comorbid conditions limiting an ultrasound evaluation of the heart. For such patients, the use of intravenous contrast agents should be encouraged as a means to provide added diagnostic information and to streamline early detection and treatment of underlying cardiac pathophysiology. As with all new technology, this document will require updates and revisions as additional data become available.

High-frequency color flow imaging of the microcirculation.

The extension of ultrasound (US) color flow imaging (CFI) techniques to high frequencies (> 20 MHz) has the potential to provide valuable noninvasive tools for scientific and clinical investigations of blood flow in the microcirculation. We describe the development of a slow-scan CFI system operating in the 20-100-MHz range that has been optimized to image the microcirculation. The apparatus has incorporated elements of a previously reported pulsed-wave Doppler system and is capable of operating in either CFI or pulsed-wave mode. The performance of the CFI system was evaluated at a center frequency of 50 MHz using two PVDF transducers with -6-dB beam widths of 43 and 60 microm. The -6 dB-axial resolutions were estimated to be 66 and 72 microm, respectively. In vivo validation experiments conducted using the murine ear model demonstrated the detection of flow in vessels down to 15-20 microm in diameter with flow velocities on the order of mm per s. Further experiments examining experimental murine tumors confirmed the successful detection of flow in the tumor microcirculation.

Pulse inversion imaging of liver blood flow: improved method for characterizing focal masses with microbubble contrast.

RATIONALE AND OBJECTIVES: To create a microbubble contrast image of vessels that lie below the resolution of an ultrasound system, a technique is required that detects preferentially the agent echo, rejecting that from tissue. Harmonic imaging exploits the nonlinear behavior of microbubbles but forces a compromise between image sensitivity and axial resolution. The authors describe and evaluate a new method that overcomes this compromise and improves contrast imaging performance: pulse inversion imaging. METHODS: Sequences of pulses of alternate phase are transmitted into tissue and their echoes summed. A prototype scanner equipped with pulse inversion was used to image phantoms and 16 patients with focal liver masses. RESULTS: Pulse inversion images show contrast sensitivity and resolution superior to that of harmonic images. Vessels can be imaged at an incident power sufficiently low to avoid destroying the agent, allowing unique visualization of tumor vasculature. Distinct patterns were seen in hemangiomas, metastases, and hepatocellular carcinomas. CONCLUSIONS: Pulse inversion imaging is an improved bubble-specific imaging method that extends the potential of contrast ultrasonography.

Infusion versus bolus of an ultrasound contrast agent: in vivo dose-response measurements of BR1.

RATIONALE AND OBJECTIVES: To determine the efficacy of an ultrasound contrast agent infusion using Doppler intensitometry estimation of backscatter enhancement in blood. METHODS: Multiple intravenous injections of BR1 (SonoVue) were performed in chronic dog studies, using bolus (0.05-2 mL) and infusion (3-40 mL/h during 6 minutes) administration. The pulsed Doppler signal from the femoral artery was recorded and analyzed for mean Doppler power and integrated fractional enhancement. RESULTS: For bolus injection, time-intensity curves exhibited a rapid first pass (peak 30 dB for 0.45 mL) followed by a slower washout. Integrated fractional enhancement exhibited a linear relation with the dose (R2 = 0.99). For infusion administration, peak enhancement increased with the infusion rate from 8 to 22 dB. At rates exceeding 30 mL/h, the enhancement was stable with a plateau-like pattern. CONCLUSIONS: Infusion of BR1 is easily achieved and allows the duration of enhancement to be increased as long as desired. Stable enhancement is obtained for rates greater than 30 mL/h.

Ultrasound for the visualization and quantification of tumor microcirculation.

Advances in ultrasound based methods for the non-invasive assessment of the tumor microcirculation are described. Two new ultrasound approaches are highlighted. The first method relies on commercially available ultrasound contrast agents in combination with specialized nonlinear imaging sequences. Nonlinear scattering by microbubble contrast agents provides a unique intravascular signature that can be distinguished from the echoes caused by surrounding tissues. Through destruction-reperfusion experiments it is possible to use microbubble contrast agents as a tracer revealing the kinetics of tumor blood flow. The second ultrasound method for examining the microcirculation involves the use of much higher frequencies. At frequencies on the order of 50 MHz, Doppler processing allows the direct assessment of flow dynamics in realtime in arterioles as small as 15 microm. Three dimensional Doppler maps of flow patterning are presented. The strengths and weaknesses of these new methods are discussed and the potential for their use in preclinical animal drug studies, clinical drug trials, and prognostic studies is described.

Harmonic hepatic US with microbubble contrast agent: initial experience showing improved characterization of hemangioma, hepatocellular carcinoma, and metastasis.

PURPOSE: To characterize blood flow in focal hepatic lesions with harmonic ultrasonographic (US) imaging and a microbubble contrast agent. MATERIALS AND METHODS: Thirty patients with known hepatic masses were examined after injection of a perfluorocarbon microbubble agent. Tumor vascularity was assessed with continuous, harmonic gray-scale imaging with a low mechanical index (MI). Tumor vascular volume was assessed with brief, high-MI insonation called interval-delay imaging, which caused microbubble destruction. As the total contrast agent volume in the liver reflects the total vascular volume, quantitation of lesion enhancement relative to normal hepatic enhancement helped determine the vascular volume of the tumor relative to that of normal parenchyma. RESULTS: Low-MI continuous harmonic imaging showed lesional vessels in hepatocellular carcinomas, minimal or no vessels in hemangiomas, and variable vascularization in metastases. High-MI interval-delay imaging showed greater enhancement in hepatocellular carcinomas than in normal liver (P <.02) and showed less enhancement in hemangiomas than in normal liver (P <.02). Enhancement in metastases was greater in the margins than in the center; as a result, the lesions appeared smaller (P <.03) and less well defined on the interval-delay images. CONCLUSION: Contrast-enhanced harmonic imaging appears superior to conventional Doppler US for hepatic mass characterization. Low-MI continuous and high-MI interval-delay imaging can help assess tumor vascular pattern and microvascular volume.

A specific sign of pneumoperitoneum on sonography: enhancement of the peritoneal stripe.

OBJECTIVE: Failure to reveal pneumoperitoneum is a recognized weakness of abdominal sonography. Our objective is to describe a reliable and reproducible sign of pneumoperitoneum that was first identified in an animal model and then confirmed in patients who had undergone laparoscopy. SUBJECTS AND METHODS: We injected 300 ml of degassed water into the peritoneal cavity of a 15-kg anesthetized pig. Sonographic images were obtained of the anterior peritoneal area after intraperitoneal injection of a single bubble, a series of bubbles, and, subsequently, a 10-ml bolus of air. Later, abdominal sonography was performed in nine patients who had undergone laparoscopy. Close attention was paid to the anterior peritoneal area and signs of free air observed in the animal model. Ten healthy volunteers functioned as a control group. RESULTS: In the pig, minute amounts of intraperitoneal air showed on sonography as enhancement of the peritoneal stripe. Larger volumes of intraperitoneal air showed as enhancement of the peritoneal stripe associated with dirty shadowing or distal multiple reflection artifacts. The stripe enhanced each time it appeared in the reflection artifact. Intraluminal gas was associated with a normal thin peritoneal stripe, superficial and distinct from the underlying gas artifact. The patients who had undergone laparoscopy showed findings suggestive of small and large pockets of free air, as we saw in the pig model. The control group showed findings consistent with intraluminal gas only. CONCLUSION: On sonography, enhancement of the peritoneal stripe alone or with reflection artifacts involving the peritoneal stripe is an accurate sign of pneumoperitoneum.

Gas at abdominal US: appearance, relevance, and analysis of artifacts.

PURPOSE: To describe the spectrum of ultrasonographic (US) appearances of intraluminal gas, including two clinically relevant gas artifacts. MATERIALS AND METHODS: Observations were made in patients and reproduced in an animal model, an ex vivo gut preparation, and a tissue-mimicking phantom. Appearances were classified according to a physical model of the interaction between sound and collections of gas. RESULTS: Free bubbles of gas appeared as bright echogenic foci extending artifactually owing to lateral and axial blooming. This causes bubbles that abut the gut wall to enhance the layer one echo, which corresponds to the interface between the mucosa and the luminal contents. Such bubbles can also falsely appear to be within the gut wall itself owing to elevation averaging and thereby cause the artifact pseudo-pneumatosis intestinalis. Isolated groups of small bubbles created a characteristic periodicity and tapering of the distal echo pattern. In the extreme case, in which many such echoes are superimposed, "dirty shadowing" occurs. A contiguous pocket of gas may cause the gut wall to appear artifactually thickened (i.e., pseudo-thickened gut). This was shown to be a form of mirror image artifact. CONCLUSION: Classification of the effects of gas on US images according to their physical characteristics may aid in their interpretation. Appreciating two previously undescribed artifacts, pseudo-pneumatosis intestinalis and pseudo-thickened gut, will improve the usefulness of abdominal US.