Martin Blomley

Microbubble contrast agents: a new era in ultrasound

Contrast agents are widely used in imaging, but until recently they had little place in ultrasonography. This has changed with the introduction of microbubbles—small (typically 3 μm in diameter) gas filled bubbles that are usually injected intravenously. Injecting a gas into the circulation may seem potentially hazardous, but extensive clinical experience has shown that the tiny volume of air or gas given (under 200 μl) is not dangerous, and the safety of microbubbles compares well to that of conventional agents in radiography and magnetic resonance imaging.1 Although microbubbles were originally designed simply to improve conventional ultrasound scanning, recent discoveries have opened up powerful emerging applications. This article describes some of these applications in radiology and cardiology and discusses the potential of microbubbles for therapy. We prepared this review from contributions from researchers with special knowledge of the use of microbubbles in radiology, cardiology, and treatment. We combined our personal experience in research over several years with a review of recent literature on the subject. Microbubbles work by resonating in an ultrasound beam, rapidly contracting and expanding in response to the pressure changes of the sound wave. By a fortunate coincidence, they vibrate particularly strongly at the high frequencies used for diagnostic ultrasound imaging. This makes them several thousand times more reflective than normal body tissues. In this way they enhance both grey scale images and flow mediated Doppler signals. As well as being useful in itself, the resonance that microbubbles produce has several special properties that can be exploited to improve diagnoses. Just as with a musical instrument, multiple harmonic signals—or overtones—are produced. Ultrasound scanners can be tuned to “listen” to these harmonics, producing strong preferential imaging of the microbubbles in an image. The selective excitation produced can also destroy microbubbles relatively easily, an effect that can be useful …

Non-invasive diagnosis of hepatic cirrhosis by transit-time analysis of an ultrasound contrast agent

BACKGROUND Hepatic cirrhosis is accompanied by several haemodynamic changes including arterialisation of the liver, intrahepatic shunts, pulmonary arteriovenous shunts, and a hyperdynamic circulatory state. We postulated that the hepatic first pass of a bolus of an ultrasound contrast agent injected into a peripheral vein is accelerated in patients with cirrhosis. We investigated this first pass in patients with diffuse liver disease and in normal controls to assess whether it provides useful differential diagnostic information. METHODS We enrolled 15 patients with biopsy-proven cirrhosis, 12 patients with biopsy-proven non-cirrhotic diffuse liver disease, and 11 normal controls. We carried out continuous spectral doppler ultrasonography of a hepatic vein from 20 s before to 3 min after a peripheral intravenous bolus injection of 2.5 g Levovist. The intensity of the doppler signal was measured and used to plot time-intensity curves. FINDINGS Patients with cirrhosis showed a much earlier onset of enhancement (arrival time; mean 18.3 s) and peak enhancement (mean 55.5 s) than controls (49.8 s and 97.5 s) or patients with non-cirrhotic diffuse liver disease (35.8 s and 79.7 s). All patients with cirrhosis had an arrival time of the bolus of less than 24 s, whereas the arrival time was 24 s or more in 22 of the 23 other participants. Peak enhancement was higher in patients with cirrhosis (mean 48.7 units) than in the other two groups (12.5 and 12.3 units, respectively). We found highly significant differences between the patients with cirrhosis and each of the other two groups for all variables (p<0.005), whereas we found no significant differences between non-cirrhotic patients and controls. INTERPRETATION Our preliminary study suggests that analysis of liver transit time of a bolus of ultrasound contrast agent provides useful information about haemodynamic changes in patients with cirrhosis. Measurement of the arrival time of the bolus allows discrimination of patients with cirrhosis from controls and from patients with non-cirrhotic diffuse liver disease, and has potential as a non-invasive test for cirrhosis.

Liver Perfusion Studied with Ultrafast CT.

Objective Our goal was to quantify absolute hepatic arterial and portal venous perfusion noninvasively in patients with and without liver disease using ultrafast CT. Materials and Methods A single slice through the porta hepatis was repeatedly scanned after bolus injection of 25 ml of iohexol 300 mg I/ml, followed by a 25 ml saline “chaser” intravenously at 10 ml/s. Thirty-nine controls, 7 cirrhotic patients, and 5 patients with known metastases on the slice plane were studied; hepatic arterial perfusion was determined in 41 patients and portal venous perfusion in 24. Time–attenuation curves from regions of interest drawn over the liver, spleen, aorta, and portal vein were analysed. Hepatic arterial perfusion was calculated by dividing the peak gradient of the liver time–attenuation curve prior to the time of peak splenic attenuation by the peak aortic CT number increase. Splenic perfusion was calculated by dividing the peak gradient of the splenic time–attenuation curve by the peak aortic CT number increase. Portal perfusion was derived by scaling the splenic time–attenuation curve by the ratio of hepatic arterial/splenic perfusion. This scaled curve was subtracted from the liver time–attenuation curve to give a portal curve. The peak up-slope of this curve was divided by the peak rise in splenic or portal vein density. Results Hepatic arterial perfusion averaged 0.19 ml/min/ml (n = 31) in controls and was raised in cirrhosis to 0.25 ml/min/ml (n = 6) and metastases 0.43 ml/min/ml (n = 4). Portal venous perfusion was 0.93 ml/min/ml (n = 19) in controls and 0.43 ml/min/ml (n = 4) in cirrhosis. Reproducibility has been confirmed. Conclusion Dynamic ultrafast CT shows potential in quantifying arterial and portal hepatic perfusion. The technique may be adaptable to dynamic bolus MRI.

Developments in ultrasound contrast media

Abstract Ultrasound microbubble contrast agents are effective and safe echo enhancers. An ingenious array of methods are employed to achieve stability and provide a clinically useful enhancement period. Microbubbles enhance ultrasound signals by up to 25 dB (greater than 300-fold increase) due to resonant behaviour. This is used to rescue failed Doppler studies and may be extended to image the microcirculation of tumours and the myocardium using non-linear modes. Functional studies open up a whole range of applications by using a variety of active and passive quantitation techniques to derive indices from the transit of contrast through a tissue of interest. This has been especially successful in the detection of liver metastases and cirrhosis and shows great promise as a clinical tool. It also has great potential in measuring microcirculatory flow velocity. The demonstration that some microbubbles are not just pure blood pool agents but have a hepatosplenic specific phase has extended the versatility of ultrasound. Imaging of this stationary phase with non-linear modes such as phase inversion and stimulated acoustic emission, has improved the sensitivity and specificity of ultrasound in the detection and characterisation of focal liver lesions to rival that of CT and MR.

Hepatic vein transit times using a microbubble agent can predict disease severity non-invasively in patients with hepatitis C

Background and aims: A reliable non-invasive assessment of the severity of diffuse liver disease is much needed. We investigated the utility of hepatic vein transit times (HVTT) for grading and staging diffuse liver disease in a cohort of patients with hepatitis C virus (HCV) infection using an ultrasound microbubble contrast agent as a tracer. Materials and methods: Eighty five untreated patients with biopsy proven HCV induced liver disease were studied prospectively. All were HCV RNA positive on polymerase chain reaction testing. Based on their histological fibrosis (F) and necroinflammatory (NI) scores, untreated patients were divided into mild hepatitis (F ⩽2/6, NI ⩽3/18), moderate/severe hepatitis (3 ⩽F <6 or NI ⩾4), and cirrhosis (F = 6/6) groups. In addition, 20 age matched healthy volunteers were studied. After an overnight fast, a bolus of contrast agent (Levovist) was injected into an antecubital vein and spectral Doppler signals were recorded from both the right and middle hepatic veins for analysis. HVTTs were calculated as the time from injection to a sustained rise in Doppler signal >10% above baseline. The Doppler signals from the carotid artery were also measured in 60 patients and carotid delay times (CDT) calculated as the difference between carotid and hepatic vein arrival times. The earliest HVTT in each patient was used for analysis. Results: Mean (SEM) HVTT for the control, mild hepatitis, moderate/severe hepatitis, and cirrhosis groups showed a monotonic decrease of 38.1 (2.8), 38.8 (2.4), 26.0 (2.4), and 15.8 (0.8) seconds, respectively. Mean (SEM) CDT for the control, mild hepatitis, moderate/severe hepatitis, and cirrhosis patients again showed progressive shortening of 30.3 (2.6), 25.9 (2.6), 14.8 (2.1), and 5.6 (1.2) seconds, respectively. There were significant differences between the groups for HVTT (ANOVA, p<0.001) and CDT (ANOVA, p<0.001). There was 100% sensitivity and 80% specificity for diagnosing cirrhosis and 95% sensitivity and 86% specificity for differentiating mild hepatitis from more severe liver disease. Conclusion: We have shown, for the first time, that HVTT using an ultrasound microbubble contrast agent can assess HCV related liver disease with clear differentiation between mild hepatitis and cirrhosis. There were significant differences between these two groups and the moderate/severe hepatitis group. CDT offers no additional benefit or greater differentiation than HVTT and can be omitted, thus simplifying this technique. HVTT may complement liver biopsy and may also be a useful alternative for assessment of liver disease in patients who have contraindications to biopsy.

Pulse-inversion mode imaging of liver specific microbubbles : improved detection of subcentimetre metastases

Pulse-inversion mode (a new ultrasound mode) can be used to image the late liver-specific parenchymal phase of the microbubble contrast-agent Levovist. Scanning in pulse-inversion mode after Levovist improves the detection of liver metastases and reveals more lesions of smaller size than conventional ultrasonography and computed tomography.

Liver microbubble transit time compared with histology and Child-Pugh score in diffuse liver disease: a cross sectional study.

Background: A previous pilot study showed that early arrival time of a microbubble in a hepatic vein is a sensitive indicator of cirrhosis. Aim: To see if this index can also grade diffuse liver disease. Patients: Thirty nine fasted patients with histologically characterised disease were studied prospectively. Nine patients had no evidence of liver fibrosis, 10 had fibrosis without cirrhosis, and 20 had cirrhosis (five Child’s A, seven Child’s B, and eight Child’s C). Methods: Bolus injections of a microbubble (Levovist; Schering, Berlin) were given intravenously, followed by a saline flush. Time intensity curves of hepatic vein and carotid artery spectral Doppler signals were analysed. Hepatic vein transit time (HVTT) was calculated as the time after injection at which a sustained signal increase >10% of baseline was seen. Carotid delay time (CDT) was calculated as the difference between carotid and hepatic vein enhancement. Results: Diagnostic studies were achieved in 38/39 subjects. Both HVTT and CDT became consistently shorter with worsening disease, as follows (means (SD)): HVTT: no fibrosis 44 (25) s, fibrosis 26 (8) s, Child’s A 21 (1) s, Child’s B 16 (3) s, and Child’s C 16 (2) s; CDT: no fibrosis 31 (29) s, fibrosis 14 (6) s, Child’s A 8 (1) s, Child’s B 4 (4) s, and Child’s C 3 (3) s. These differences were highly significant (p<0.001, ANOVA comparison). A HVTT <24 s and a CDT <10 s were 100% sensitive for cirrhosis (20/20 and 18/18, respectively) but not completely specific: 2/8 subjects with fibrosis had CDT values <10 s and 3/9 had HVTT <24 s. Conclusion: This minimally invasive test shows promise not only in diagnosing cirrhosis but also in assessing disease severity.

Multi-centre clinical study evaluating the efficacy of SonoVue™ (BR1), a new ultrasound contrast agent in Doppler investigation of focal hepatic lesions

OBJECTIVES SonoVue is a new ultrasound contrast agent, which consists of stabilised microbubbles of a sulphur hexafluoride gas. The aim of the study was to assess its efficacy in the Doppler investigation of focal hepatic lesions. MATERIALS AND METHODS Seventy patients with focal liver tumours were studied. Four doses (0.3, 0.6, 1.2 and 2.4 ml) of SonoVue were administered intravenously with at least 10 min delay between each injection. A complete colour/power and spectral Doppler imaging investigation of the lesions was performed at baseline pre-dosing and after each SonoVue injection. All examinations were recorded on SVHS videotapes. Baseline and post contrast videotapes were reviewed by the on-site (un-blinded) investigators and by two off-site blinded readers (a) to grade the global quality of the Doppler scans of the focal lesions vascularity and the normal parenchymal vessels (b) to measure the duration of clinically useful Doppler signal enhancement and (c) to determine the diagnostic accuracy and performance of the enhanced versus unenhanced scans using histopathology, tumour markers, CT and/or MR as the reference standard. RESULTS A statistically significant improvement was observed at all four SonoVue doses in the off site assessment of global quality of the Doppler examination of tumoral and normal parenchymal vessels in comparison with the baseline (P < 0.05). The median duration of clinically useful enhancement was significantly increased with increasing doses (P < 0.001), ranging between 1.4-2.2 min for the lowest dose and 3.2-3.8 min for the highest dose for the off-site readers. On-site assessment of diagnostic accuracy showed a significant increase in the specificity of the Doppler diagnoses (P < 0.0016) with an increase in the positive and negative predictive values and in the likelihood ratio in differentiating between benign and malignant lesions. Off-site evaluation showed a significant increase in the accuracy of enhanced Doppler diagnosis in comparison with the baseline performance. CONCLUSION The results suggest that SonoVue is effective in improving the display of tumoral vascularisation and may be useful in the characterisation of focal liver lesions.

Quantification of blood flow

Abstract. Traditionally, Doppler ultrasound has been used to estimate blood flow as the mean velocity multiplied by the vessel area, but this is subject to significant errors and may be difficult to perform accurately. Microbubbles, developed as contrast agents for ultrasound, were initially envisaged as useful for increasing the intensity of echoes and thus rescuing Doppler studies that were technical failures because of attenuated signals or very slow flow. However, they can act as tracers and, by analogy with isotope techniques, can be used to measure blood flow with transit-time methods which exploit both arterial and venous time–intensity data. An acceptable compromise is to acquire both a tissue intensity curve and one from the feeding artery. The transit of microbubbles across an organ or tissue can be used to estimate haemodynamic alterations, e.g. the arterialisation of the supply to the liver in malignancies and cirrhosis and the delayed arterio-venous transit in the transplant kidney during rejection. The fragility of microbubbles can be turned to advantage by being exploited to create a negative bolus by exposing a tissue slice to a high power beam. The rate of refilling of this slice by circulating microbubbles can then be followed with a low-intensity monitoring beam and the resulting rising exponential curve analysed to extract indices of both the reperfusion rate (the slope) and the fractional vascular volume (the asymptote). The product of these is a measure of true tissue perfusion.

Contrast Bolus Dynamic Computed Tomography for the Measurement of Solid Organ Perfusion

RATIONALE AND OBJECTIVES. The authors have investigated the aortic responses to various intravenous bolus injections of nonionic and ionic contrast media and have presented data illustrating the potential of ultrafast computed tomography (CT) to quantify perfusion in the kidney, liver, and spleen.METHODS. Bolus Dynamics Study: Performed in 3 healthy dogs (weight: 35kg to 36 kg). In 2 dogs, 15 mL of the nonionic agent iohexol and the ionic agent sodium-meglumine diatrizoate were injected at 5, 10 and 20 mL/sec via a venous catheter placed in the superior vena cava; the order of injection was alternated between the 2 dogs. In the third dog, 25 mL of iohexol 300 mg I/mL was compared with diatrizoate 370 mg I/mL with injection rates of 10 and 20 mL/sec. Computed tomography scanning at the level of the midabdominal aorta was performed using an ultrafast CT scanner. Time-density curves were drawn for regions of interest over the aorta, and gamma-variate fits performed. Perfusion Studies: Dynamic perfusion scans of the upper abdomen were performed in more than 50 patients. A dose of 25 mL of iohexol 300 mg I/mL was injected at 10 mL/sec via an intravenous cannula in the ante-cubital fossa, followed immediately by 25 mL of saline, at the same rate. Scanning was performed at a single level using an ultrafast CT scanner. Regions of interest were drawn and gamma-variate fits were applied to the vascular time-density curves.RESULTS. Bolus Dynamics: Excellent curve fits for aortic time-density curves were obtained. A 10-mL/sec versus a 5-mL/sec bolus produced an 8% higher peak density. Nonionic contrast increased the peak density by a mean of 6%, increased the area under the corrected time-density curve by a mean of 22%, and lengthened the increase time by a mean of 21%. Perfusion Studies: Values obtained were reproducible and correlated well with values predicted from inert gas washout techniques.CONCLUSIONS. Changes in the CT number in a region after an intravenous injection of contrast medium may be used to calculate blood flow per unit volume of tissue. Ultrafast CT offers sufficient data points for accurate calculation. The quality of the aortic bolus is of great importance. Nonionic media offer several important advantages: hemodynamic perturbation is minimized, and they are better tolerated at the high injection rates needed. Low-osmolality nonionic agents produce “better” curves than conventional high-osmolality ionic agents, all other factors being equal. The resulting data are relevant to intravenous digital subtraction angiography and indirect portography as well as to perfusion measurement. The technique of quantitative dynamic CT is theoretically applicable to any cross-sectional modality, notably magnetic resonance.