Use of ultrasound pulses combined with Definity for targeted blood-brain barrier disruption: a feasibility study

We have developed a method to use low-intensity focused ultrasound pulses combined with an ultrasound contrast agent to produce temporary blood-brain barrier disruption (BBBD). This method could provide a means for the targeted delivery of drugs or imaging agents into the brain. In all our previous work, we used Optison as the ultrasound contrast agent. The purpose of this study was to test the feasibility of using the contrast agent Definity for BBBD. A total of 36 non-overlapping locations were sonicated through a craniotomy in experiments in the brains of nine rabbits (four locations per rabbit; ultrasound [US] frequency: 0.69 MHz; burst: 10 ms; pulse repetition frequency (PRF): 1 Hz; duration: 20 s). The peak negative pressure amplitude ranged from 0.2 to 1.5 MPa. An additional 11 locations were sonicated using Optison at pressure amplitude of 0.5 MPa. Definity and Optison dosages were the same as those used clinically for ultrasound imaging: 10 and 50 microl/kg, respectively. The probability for BBBD (determined using MRI contrast agent enhancement) as a function of pressure amplitude was similar to that found earlier with Optison. For both agents, the probability was estimated to be 50% at 0.4 MPa using probit regression. Histologic examination revealed small, isolated areas of extravasated erythrocytes in some locations. At 0.8 MPa and higher, these areas were sometimes accompanied by tiny (dimensions of 100 microm or less) regions of damaged brain parenchyma. The magnitude of the BBBD was larger with Optison than with Definity at 0.5 MPa (signal enhancement: 13.3% +/- 4.4% vs. 8.4% +/- 4.9%; p = 0.04). In addition, more areas with extravasated erythrocytes were observed with Optison (5.0 +/- 3.5 vs. 1.4 +/- 1.9 areas with extravasation in histology section with largest effect; p = 0.03). We concluded that BBBD is possible using Definity at the dosage of contrast agent and the acoustic parameters tested in this study. The probability for BBBD as a function of pressure amplitude and the type of acute tissue effects were similar to what has been observed using Optison. However, under the experimental conditions used in this study, Optison produced a larger effect for the same acoustic pressure amplitude.     

Creating Brain Lesions with Low-Intensity Focused Ultrasound with Microbubbles: A Rat Study at Half a Megahertz

Low-intensity focused ultrasound was applied with microbubbles (Definity, Lantheus Medical Imaging, North Billerica, MA, USA; 0.02 mL/kg) to produce brain lesions in 50 rats at 558 kHz. Burst sonications (burst length: 10 ms; pulse repetition frequency: 1 Hz; total exposure: 5 min; acoustic power: 0.47–1.3 W) generated ischemic or hemorrhagic lesions at the focal volume revealed by both magnetic resonance imaging and histology. Shorter burst time (2 ms) or shorter sonication time (1 min) reduced the probability of lesion production. Longer pulses (200 ms, 500 ms and continuous wave) caused significant near-field damage. Using microbubbles with focused ultrasound significantly reduced acoustic power levels and, therefore, avoided skull heating issues and potentially can extend the treatable volume of transcranial focused ultrasound to brain tissues close to the skull.     

磁共振引导聚焦超声在中枢神经系统疾病中的应用

磁共振引导聚焦超声是一种先进的无创治疗技术, 在通过靶向热消融及开放血脑屏障治疗中枢神经系统疾病方面具有较大潜力, 目前已被应用于特发性震颤、帕金森病、颅内肿瘤、阿尔兹海默病、强迫症、神经病理性疼痛等疾病的研究与治疗中。磁共振引导聚焦超声因其精准、无电离辐射、实时监测靶点温度等优势可能成为诸多疾病的潜在替代疗法, 为患者提供一种新的治疗选择。本文就目前磁共振引导聚焦超声在中枢神经系统疾病方面的临床应用及科学研究予以综述。     

A review of magnetic resonance imaging-guided focused ultrasound surgery of uterine fibroids

Uterine fibroids are a significant cause of morbidity for women of reproductive age. Over the past decade, minimally invasive treatment options are becoming increasingly popular. A new, Food and Drug Administration-approved noninvasive treatment option is magnetic resonance-guided focused ultrasound surgery, which has the potential to become a treatment of choice for selected patients. We review the technical aspects of the procedure of magnetic resonance-guided focused ultrasound surgery for treatment of uterine fibroids, potential difficulties with treatment planning, and clinical trial results to date. We also describe current developments in treatment imaging and treatment optimization.     

New clinical applications of magnetic resonance-guided focused ultrasound

Magnetic resonance-guided ultrasound delivers destructive energy into deep body structures with great accuracy and repeatability with an excellent safety profile. The use of this technology for the treatment of uterine fibroids is already becoming widespread. This article reviews the further areas of magnetic resonance-guided focused ultrasound application that are evolving and how they will be applied to other parts of the body.     

Combination of thermal and cavitation effects to generate deep lesions with an endocavitary applicator using a plane transducer: ex vivo studies

In the high-intensity focused ultrasound (US), or HIFU, field, it is well-known that the cavitation effect can be used to induce lesions of larger volume. The principle is based on the increase in the equivalent attenuation coefficient of the tissue in the presence of the bubbles created by cavitation. The elementary lesions produced by combination of cavitation and thermal effects, using focused transducers, were spherical and developed upstream of the focal point. This paper presents a method that combines cavitation with a thermal effect to obtain deeper lesions using a plane transducer, rather than a focused one. The cavitation effect was produced by delivering intensities of 60 W/cm2 at the face of the transducer for 0.5 s. The applicator was then rotated through 90 degrees at a constant speed of between 0.5 and 1.5 degrees /s. During this rotation, ex vivo tissues were exposed continuously to an acoustic intensity of 14 W/cm2 to combine the cavitation effect with a thermal effect. The necroses were, on average, twice as deep when the cavitation effect was used as those obtained with a thermal effect alone. Observed macroscopically, the lesions have a very well-delimited geometry. Temperature measurements made at different angles of treatment have shown that they were coagulation necroses.     

Pre-Clinical Study of In Vivo Magnetic Resonance-Guided Bubble-Enhanced Heating in Pig Liver

Bubble-enhanced heating (BEH) can be exploited to increase heating efficiency in treatment of liver tumors with non-invasive high-intensity focused ultrasound (HIFU). The objectives of this study were: (i) to demonstrate the feasibility of increasing the heating efficiency of sonication exploiting BEH in pig liver in vivo using a clinical platform; (ii) to determine the acoustic threshold for such effects with real-time, motion-compensated magnetic resonance-guided thermometry; and (iii) to compare the heating patterns and thermal lesion characteristics resulting from continuous sonication and sonication including a burst pulse. The threshold acoustic power for generation of BEH in pig liver in vivo was determined using sonication of 0.5-s duration (“burst pulse”) under real-time magnetic resonance thermometry. In a second step, experimental sonication composed of a burst pulse followed by continuous sonication (14.5 s) was compared with conventional sonication (15 s) of identical energy (1.8 kJ). Modification of the heating pattern at the targeted region located at a liver depth between 20 and 25 mm required 600–800 acoustic watts. The experimental group exhibited near-spherical heating with 40% mean enhancement of the maximal temperature rise as compared with the conventional sonication group, a mean shift of 7 ± 3.3 mm toward the transducer and reduction of the post-focal temperature increase. Magnetic resonance thermometry can be exploited to control acoustic BEH in vivo in the liver. By use of experimental sonication, more efficient heating can be achieved while protecting tissues located beyond the focal point.     

The threshold for thermally significant cavitation in dog's thigh muscle in vivo

In this study the threshold of thermally significant transient cavitation in vivo in dog's thigh muscle was investigated as a function of frequency from 0.246 MHz to 1.68 MHz. Cavitation, evidenced by strong emission of wide band noise monitored by a hydrophone, appeared to increase the energy absorption in tissue at the focal zone of a focused ultrasound beam as measured with an embedded thermocouple. This was indicated by a significant increase in the temperature, a loss of smooth temperature rise during the 1 s sound pulse and a significant reduction in the acoustic power transmitted through the thigh. This thermal phenomenon was associated with a strong emission of wide band noise which was monitored by a hydrophone. In addition, strong echoes appeared in ultrasound images during the pulses that caused the noise emission and the thermal effect. These echoes appeared preferentially at locations where there was acoustic heterogeneity. The measured cavitation pressure amplitude threshold was found to depend almost linearly on frequency with a slope of about 5.3 MPa MHz-1. (The extrapolated static pressure threshold was 0.6 MPa). When these measured levels are compared to those typical of clinical application, it appears that the transient cavitation can be avoided when perfusion independent high temperature hyperthermia is induced with focused and pulsed ultrasound fields. However, intensities required during scanned focused ultrasound hyperthermia, where sharply focused transducers are used to heat large tumors at low frequencies (1 MHz or below), could rise above the threshold. Thus, care should be taken when focused ultrasound systems are designed so that the maximum peak pressure is below the threshold in order to avoid unpredictable biological effects induced by transient cavitation. Finally it is unlikely that the present diagnostic ultrasound units which operate at higher frequencies and in pulsed mode could cause transient cavitation in vivo.     

Simulation of Intracranial Acoustic Fields in Clinical Trials of Sonothrombolysis

Two clinical trials have used ultrasound to improve tPA thrombolysis in patients with acute ischemic stroke. The Combined Lysis of Thrombus in Brain Ischemia Using Transcranial Ultrasound and Systemic tPA (CLOTBUST) trial reported accelerated recanalisation of the middle cerebral artery (MCA) in patients with symptoms of MCA infarction, which were monitored with 2-MHz transcranial Doppler. In CLOTBUST, there was no increased bleeding as evidenced by cranial computed tomography. The Transcranial Low-Frequency Ultrasound-Mediated Thrombolysis in Brain Ischemia (TRUMBI) trial, which employed magnetic resonance imaging (MRI) before and after tPA thrombolysis, was discontinued prematurely because of an increased number of secondary hemorrhages, possibly related to the use of low frequency 300-kHz ultrasound. The purpose of our work is to help identify possible mechanisms of intracerebral hemorrhage resulting from sonothrombolysis by applying a simulation tool that estimates the pressure levels in the human brain that are produced with different sonothrombolysis devices. A simulation software based on a finite difference time domain (FDTD) three-dimensional (3D) scheme was developed to predict acoustic pressures in the brain. This tool numerically models the wave propagation through the skull and reproduces both ultrasound protocols of CLOTBUST and TRUMBI for analysis of the distribution of acoustic pressure in the brain during stroke treatment. For the simulated TRUMBI trial, we analyzed both a “high” and “low” hypothesis according to published parameters (for high and low amplitude excitations). For these hypotheses, the mean peak rarefactional pressures in the brain were 0.26 ± 0.2 MPa (high hypothesis) and 0.06 ± 0.05 MPa (low hypothesis), with maximal local values as high as 1.2 MPa (high hypothesis) and 0.27 MPa (low hypothesis) for configurations modelled in this study. The peak rarefactional pressure was thus higher than the inertial acoustic cavitation threshold in the presence of a standing wave in large areas of the brain, even outside the targeted clot. For the simulated CLOTBUST trial, the maximum peak negative pressure was less than 0.07 MPa. This simulated pressure is below the threshold for both inertial and stable acoustic cavitation but likewise lower than any acoustic pressure that has been reported as sufficient for effective sonothrombolysis. Simulating the pressure field of ultrasound protocols for clinical trials of sonothrombolysis may help explain mechanisms of adverse effects. Such simulations could prove useful in the initial design and optimization of future protocols for this promising therapy of ischemic stroke. (E-mail: norabelic@yahoo.fr)     

Flash effect of contrast microbubbles by ultrasound exposure

Background Microbubbles used in contrast echo examination are destroyed by exposure to ultrasound but develop a new ultrasound wave on destruction; the so-called flash effect. Factors influencing the magnitude of the new wave have yet to be elucidated. Here we investigate the method of assessing this effect and attempt to clarify the relevant differences between contrast agents. Methods Three contrast agents were used: Albunex, Optison (FS 069), and Levovist. We used fundamental mode (3.75 MHz) and harmonic mode (2.5 to 5.0 MHz) ultrasound produced by a prototype echocardiograph (Toshiba) and measured the video intensity (VI) (256 gray scale) of each contrast agent contained in a thin rubber sack while changing acoustic power from a minimum level to high levels of +10.5 dB +16.5 dB, and +22.5 dB. Results VI was not changed by low acoustic power; however, it increased rapidly for a short time and then decreased rapidly when exposed to high acoustic power. The increase in VI varied with acoustic power: 30 to 60 at +10.5 dB and 70 to 115 at +22.5 dB. The increase in VI was larger in harmonic mode than in fundamental mode. The degree of decrease in VI after the flash effect correlated with the extent of increase in VI produced by the flash effect. Conclusions The flash effect occurred with each of the contrast agents, and its magnitude varied with acoustic power and contrast agent.     

Measurement of acoustic streaming using magnetic resonance

Magnetic resonance imaging (MRI) has been used to explore acoustic streaming caused in water under ultrasonic exposure conditions similar to those used for diagnostic applications. Streaming was established in an enclosed tube with acoustically transparent end windows, using a pulsed, weakly-focused transducer of acoustic frequency 3.5 MHz. Phase-detection MRI was used to image and quantify streaming profiles in the region of the acoustic focus. Acoustic powers in the range 0.4 mW to 100 mW were used. The sensitivity of the technique enabled streaming velocities down to 0. 1 mm s(-1) to be measured, generated by acoustic power less than 1 mW. In addition, acoustic streaming generated within open meshes with minimum pore dimensions of 3.0 mm and 2.0 mm was measured. The flow velocity in the coarser mesh reached 0.9 mm s(-1) at 95 mW total acoustic power. These observations demonstrate that acoustic streaming is probably a much more general phenomenon in diagnostic ultrasound (ultrasound) than previously recognised. The combination of magnetic resonance and ultrasound shows promise as a diagnostic method for the differentiation of cystic lesions in vivo, and for their characterisation, with sensitivity significantly greater than using ultrasound alone.     

Noninvasive, transcranial and localized opening of the blood-brain barrier using focused ultrasound in mice

The feasibility of blood-brain barrier (BBB) opening in the hippocampus of wild-type mice using focused ultrasound (FUS) through the intact skull and skin was investigated. Needle hydrophone measurements through ex vivo skulls revealed minimal attenuation ( approximately 18% of the pressure amplitude), a well-focused beam pattern and minute focus displacement through the parietal bone. In experiments in vivo, the brains of three mice were sonicated transcranially. Pulsed ultrasound sonications at 1.5 MHz and acoustic pressures ranging from 0.8 to 2.7 MPa were used at 20% duty cycle. Before sonication, a bolus of 10 microL of an ultrasound contrast agents (Optison) was injected intravenously. Contrast-enhanced high-resolution magnetic resonance imaging (9.4 T) revealed BBB opening and allowed for the monitoring of the slow permeation of gadolinium in the hippocampus. The region of the brain where BBB opening occurred increased with the pressure amplitude. These findings thus demonstrated the feasibility of locally opening the BBB in mice using FUS through intact skull and skin and serve as the first step in determining and assessing feasibility of drug delivery to specific regions in the mouse brain using FUS.     

Magnetic Resonance Imaging for the Exploitation of Bubble-Enhanced Heating by High-Intensity Focused Ultrasound: A Feasibility Study in ex Vivo Liver

Bubble-enhanced heating (BEH) may be exploited to improve the heating efficiency of high-intensity focused ultrasound in liver and to protect tissues located beyond the focal point. The objectives of this study, performed in ex vivo pig liver, were (i) to develop a method to determine the acoustic power threshold for induction of BEH from displacement images measured by magnetic resonance acoustic radiation force imaging (MR-ARFI), and (ii) to compare temperature distribution with MR thermometry for HIFU protocols with and without BEH. The acoustic threshold for generation of BEH was determined in ex vivo pig liver from MR-ARFI calibration curves of local tissue displacement resulting from sonication at different powers. Temperature distributions (MR thermometry) resulting from “conventional” sonications (20 W, 30 s) were compared with those from “composite” sonications performed at identical parameters, but after a HIFU burst pulse (0.5 s, acoustic power over the threshold for induction of BEH). Displacement images (MR-ARFI) were acquired between sonications to measure potential modifications of local tissue displacement associated with modifications of tissue acoustic characteristics induced by the burst HIFU pulse. The acoustic threshold for induction of BEH corresponded to a displacement amplitude of approximately 50 μm in ex vivo liver. The displacement and temperature images of the composite group exhibited a nearly spherical pattern, shifted approximately 4 mm toward the transducer, in contrast to elliptical shapes centered on the natural focal position for the conventional group. The gains in maximum temperature and displacement values were 1.5 and 2, and the full widths at half-maximum of the displacement data were 1.7 and 2.2 times larger than in the conventional group in directions perpendicular to ultrasound propagation axes. Combination of MR-ARFI and MR thermometry for calibration and exploitation of BEH appears to increase the efficiency and safety of HIFU treatment.     

Myocardial Thermal Ablation with a Transesophageal High-Intensity Focused Ultrasound Probe: Experiments on Beating Heart Models

Described here is a study of transesophageal thermal ablation of isolated and perfused beating hearts and non-human primates. An endoscope integrating a transesophageal echocardiography probe and a high-intensity focused ultrasound transducer was built and tested on five Langendorff-isolated hearts and three 30-kg baboons. B-Mode ultrasound, passive elastography and magnetic resonance imaging were performed to monitor thermal lesions. In isolated hearts, continuous and gated sonication parameters were evaluated with acoustic intensities of 9–12 W/cm². Sonication parameters of gated exposures with 12 W/cm² acoustic intensity for 5 min consistently produced visible lesions in the ventricles of isolated hearts. In animals, left atria and ventricles were exposed to repeated continuous sonications (4–15 times for 16 s) at an acoustic intensity at the surface of the transducer of 9 W/cm². Clinical states of the baboons during and after the treatment were good. One suspected lesion in the left ventricle could be evidenced by elastography, but was not confirmed by magnetic resonance imaging. The transesophageal procedure therefore has the potential to create thermal lesions in beating hearts and its safety in clinical practice seems promising. However, further technical exploration of the energy deposition in the target would be necessary before the next pre-clinical experiments.     

Effects of pulsed ultrasound on ocular tissue

Focused ultrasound at a center frequency of 9.8 MHz was used to create lesions of the retina and choroid in the proptosed eye of the anesthetized albino rabbit. Pulsed ultrasound was employed and results were compared to those obtained under continuous-wave (CW) exposures. Short pulses (e.g. 100 μsec) delivered at high repetition frequencies (e.g. 3 KHz) produced the same average intensity threshold values as those found for CW conditions. Longer pulses (e.g. 0.1 sec) delivered at low repetition frequencies (e.g. 2 Hz) produced lesions at lower temporally averaged intensities. The lowered thresholds are related to cyclical blanching occurring in insonified choroidal blood vessels. All lesion-producing intensities (approximately 100 W/cm²) were orders of magnitude larger than diagnostic levels.     

Thermal Fixation of Swine Liver Tissue after Magnetic Resonance-Guided High-Intensity Focused Ultrasound Ablation

The aim of this study was to investigate experimental conditions for efficient and controlled in vivo liver tissue ablation by magnetic resonance (MR)-guided high-intensity focused ultrasound (HIFU) in a swine model, with the ultimate goal of improving clinical treatment outcome. Histological changes were examined both acutely (four animals) and 1 wk after treatment (five animals). Effects of acoustic power and multiple sonication cycles were investigated. There was good correlation between target size and observed ablation size by thermal dose calculation, post-procedural MR imaging and histopathology, when temperature at the focal point was kept below 90°C. Structural histopathology investigations revealed tissue thermal fixation in ablated regions. In the presence of cavitation, mechanical tissue destruction occurred, resulting in an ablation larger than the target. Complete extra-corporeal MR-guided HIFU ablation in the liver is feasible using high acoustic power. Nearby large vessels were preserved, which makes MR-guided HIFU promising for the ablation of liver tumors adjacent to large veins.     

MR引导下聚焦超声术治疗子宫肌瘤的研究进展

当前已有多种临床手段治疗子宫肌瘤,但大多数为侵入性方法。近年来,MR引导下聚焦超声术(MRgFUS)作为子宫肌瘤的一种微创治疗方法在国外已成研究热点,且应用前景广阔,而在国内报道甚少,作者对这一新技术进行综述。     

Thermal effects of focused ultrasound energy on bone tissue.

The effects of focused ultrasound (US) at therapeutic acoustic power levels were studied in vivo on the bone-muscle interface in rabbit thighs. The purpose of this study was to provide direction in establishing safety guidelines for treating tissue masses using focused US on or near bone. A positioning device was used to manipulate a focused US transducer (1.5 MHz) in a magnetic resonance imaging (MRI) scanner. This system was used to sonicate the femurs of 10 rabbits at acoustic power levels of 26, 39, 52 and 65 W for 10 s. The rabbits were euthanized either 4 h or 28 days after the sonications and the bone samples were harvested for histology examinations. In the femurs studied, acoustic power levels from 39 to 65 W resulted in soft tissue damage characterized grossly by coagulated tissue and bone damage depicted by yellow discoloration. Histologic examination of lesions from sonications from 39 to 65 W demonstrated that osteocyte damage and necrosis, characterized by pyknotic cells and empty lacunae, occurred within the ablation area extending through the bone. The follow-up MR images demonstrated an increase in the amount of damage in the femurs at 28 days posttreatment in comparison to images taken immediately after treatment. Focused US directed at the femur caused immediate significant thermal damage to bone in the form of osteocyte necrosis extending through the (approximately) 1 cm bone in this study. The results suggest that, when focused US energy is directed at or near bone-muscle interfaces, precautions should be taken to avoid thermal damage to the bone that can compromise its strength for extended periods.     

Depletion of hepatic UDP-glucuronic acid by drugs that are glucuronidated

Salicylamide, clofibric acid, valproic acid and chloramphenicol are all known to be glucuronidated. The effects of these compounds on the hepatic concentration of UDP-glucuronic acid, the cosubstrate for glucuronidation, were studied in mice and found to lower hepatic UDP-glucuronic acid in a dose- and time-dependent fashion. Valproic acid, chloramphenicol, salicylamide and clofibric acid depleted hepatic UDP-glucuronic acid significantly at dosages as low as 0.5, 0.5, 0.75 and 4.0 mmol/kg, respectively. Hepatic UDP-glucuronic acid was decreased by 90% by valproic acid, 91% by chloramphenicol, 98% by salicylamide and 41% by clofibric acid (after dosages of 4, 2, 1 and 5 mmol/kg, respectively). UDP-glucuronic acid was depleted maximally by 15 after drug administration. Salicylamide also was used as a model compound to study the effect of drug loading on the hepatic concentrations of UDP-glucose and glycogen, precursors of UDP-glucuronic acid. It was found that, in addition to UDP-glucuronic acid, salicylamide (4 mmol/kg) also depleted UDP-glucose and glycogen by about 50%. These data suggest that large drug loads cause an increase flux through the glucuronic acid pathway and that hepatic UDP-glucuronic acid is consumed more rapidly than it is produced.