Focused ultrasound delivery of a selective TrkA agonist rescues cholinergic function in a mouse model of Alzheimer's disease.

The degeneration of cholinergic neurons is a prominent feature of Alzheimer's disease (AD). In animal models of injury and aging, nerve growth factor (NGF) enhances cholinergic cell survival and function, contributing to improved memory. In the presence of AD pathology, however, NGF-related therapeutics have yet to fulfill their regenerative potential. We propose that stimulating the TrkA receptor, without p75NTR activation, is key for therapeutic efficacy. Supporting this hypothesis, the selective TrkA agonist D3 rescued neurotrophin signaling in TgCRND8 mice, whereas NGF, interacting with both TrkA and p75NTR, did not. D3, delivered intravenously and noninvasively to the basal forebrain using MRI-guided focused ultrasound (MRIgFUS)-mediated blood-brain barrier (BBB) permeability activated TrkA-related signaling cascades and enhanced cholinergic neurotransmission. Recent clinical trials support the safety and feasibility of MRIgFUS BBB modulation in AD patients. Neuroprotective agents targeting TrkA, combined with MRIgFUS BBB modulation, represent a promising strategy to counter neurodegeneration in AD.

Ultrasound-responsive droplets for therapy: A review.

The last two decades have seen the development of acoustically activated droplets, also known as phase-change emulsions, from a diagnostic tool to a therapeutic agent. Through bubble effects and triggered drug release, these superheated agents have found potential applications from oncology to neuromodulation. The aim of this review is to summarise the key developments in therapeutic droplet design and use, to discuss the current challenges slowing clinical translation, and to highlight the new frontiers progressing towards clinical implementation. The literature is summarised by addressing the droplet design criteria and by carrying out a multiparametric study of a range of droplet formulations and their associated vaporisation thresholds.

Gene delivery to the spinal cord using MRI-guided focused ultrasound.

Non-invasive gene delivery across the blood-spinal cord barrier (BSCB) remains a challenge for treatment of spinal cord injury and disease. Here, we demonstrate the use of magnetic resonance image-guided focused ultrasound (MRIgFUS) to mediate non-surgical gene delivery to the spinal cord using self-complementary adeno-associated virus serotype 9 (scAAV9). scAAV9 encoding green fluorescent protein (GFP) was injected intravenously in rats at three dosages: 4 × 10(8), 2 × 10(9) and 7 × 10(9) vector genomes per gram (VG g(-1)). MRIgFUS allowed for transient, targeted permeabilization of the BSCB through the interaction of focused ultrasound (FUS) with systemically injected Definity lipid-shelled microbubbles. Viral delivery at 2 × 10(9) and 7 × 10(9) VG g(-1) leads to robust GFP expression in FUS-targeted regions of the spinal cord. At a dose of 2 × 10(9) VG g(-1), GFP expression was found in 36% of oligodendrocytes, and in 87% of neurons in FUS-treated areas. FUS applications to the spinal cord could address a long-term goal of gene therapy: delivering vectors from the circulation to diseased areas in a non-invasive manner.

The targeting accuracy of a preclinical MRI-guided focused ultrasound system.

PURPOSE: Assess the accuracy, precision, and sources of error using a preclinical MR-guided focused ultrasound system. METHODS: A preclinical focused ultrasound system, described previously [Chopra et al., Med. Phys. 36(5), 1867-1874 (2009)], was tested on a benchtop and with 3T GE, 3T Philips, and 7 T Bruker MR scanners for spatial targeting accuracy and precision. Randomly distributed water-filled holes drilled into a polystyrene plate were imaged using MRI and targeted using treatment planning software. The ultrasound focus of a 72 mm, f-number 0.8, 1.16 MHz transducer was aimed at the target locations, and 1-2 s continuous-wave sonications were performed on clear polystyrene plates to create localized spots of melted plastic. The distance between target and observed locations was measured and analyzed. Retrospective analysis of targeting accuracy was performed on preclinical data obtained from other experiments at their institution using the same system. RESULTS: The results suggest that the sources of targeting error under MR guidance can be roughly separated into three components--normally distributed random error; constant shift from inaccuracy in detection of the initial ultrasound focus; and angular misalignment between MR and focused ultrasound (FUS) coordinates. The lower bound on the targeting error was estimated to be 0.25 ± 0.13 mm, while the maximum observed targeting error did not exceed 2 mm. Measures required to reduce errors and improve targeting were developed to reduce the registration and misalignment errors such that maximum error was reduced to 0.36 ± 0.14 mm. Retrospective in vivo analysis indicated that the error was 1.02 ± 0.43 mm, including error extrinsic to the system. CONCLUSIONS: The FUS system, as described, is capable of precise and accurate sonications. The largest source of error--misregistration of the coordinate systems of the scanner and ultrasound system--was addressed which reduced the error to 0.36 ± 0.14 mm, sufficient for many preclinical applications.

Modeling the pattern of contrast extravasation in acute intracerebral hemorrhage using dynamic contrast-enhanced MR.

BACKGROUND: Contrast extravasation (CE) in spontaneous intracerebral hemorrhage (ICH), coined the spot sign, predicts hematoma expansion (HE) and poor clinical outcome. The dynamic relationship between CE and the mode of ICH growth are poorly understood. We characterized the in vivo pattern and rate of HE using a novel animal model of acute ICH. METHODS: Basal ganglia ICH was created in 14 Yorkshire swine utilizing a novel MRI integrated model, permitting real-time CE observation using dynamic contrast-enhanced (DCE) MRI. Computerized planimetry measured CE volume at each time point. Spatial vector analysis along three orthogonal axes determined distance vectors. Maximizing and minimizing the coefficient of determination defined the temporal phases of growth and stability, respectively. CE rate was calculated using a Patlak model. RESULTS: Asymmetric growth and variable rates of expansion characterized HE defining three distinct growth phases and patterns. A primary growth phase (duration 160 s; IQR 50-130) demonstrated rapid linear growth (0.04 mm/s IQR 0.01-0.10) accounting for 85 ± 15 % of total HE. The stationary phase demonstrated stability (duration 145 s; IQR 0-655). A secondary growth phase (duration 300; 130-600 s) accounted for 23 ± 8 % of total HE. In the primary and secondary growth phase, asymmetric growth occurred in the anterior-posterior (AP) planes (0.056 mm/s; p = 0.026 and 0.0112 mm/s; p = 0.03). Monophasic 2 (14 %), biphasic 4 (35 %) (primary followed by secondary growth), and triphasic 8 (56 %) patterns (primary, stationary, and secondary growth phase) were observed. CONCLUSIONS: A novel model of ICH provides real-time study of the dynamics and rate of CE. This data facilitates the understanding of pattern and rate of ICH formation.

An in vivo, MRI-integrated real-time model of active contrast extravasation in acute intracerebral hemorrhage.

BACKGROUND AND PURPOSE: The "spot sign" or contrast extravasation is strongly associated with hematoma formation and growth. An animal model of contrast extravasation is important to test existing and novel therapeutic interventions to inform present and future clinical studies. The purpose of this study was to create an animal model of contrast extravasation in acute intracerebral hemorrhage. MATERIALS AND METHODS: Twenty-eight hemispheres of Yorkshire male swine were insonated with an MR imaging-guided focused sonography system following lipid microsphere infusion and mean arterial pressure elevation. The rate of contrast leakage was quantified by using dynamic contrast-enhanced MR imaging and was classified as contrast extravasation or postcontrast leakage by using postcontrast T1. Hematoma volume was measured on gradient recalled-echo MR imaging performed 2 hours postprocedure. Following this procedure, sacrificed brain was subjected to histopathologic examination. Power level, burst length, and blood pressure elevation were correlated with leakage rate, hematoma size, and vessel abnormality extent. RESULTS: Median (intracerebral hemorrhage) contrast extravasation leakage was higher than postcontrast leakage (11.3; 6.3-23.2 versus 2.4; 1.1-3.1 mL/min/100 g; P<.001). Increasing burst length, gradient recalled-echo hematoma (ρ=0.54; 95% CI, 0.2-0.8; P=.007), and permeability were correlated (ρ=0.55; 95% CI, 0.1-0.8; P=.02). Median permeability (P=.02), gradient recalled-echo hematoma (P=.02), and dynamic contrast-enhanced volumes (P=.02) were greater at 1000 ms than at 10 ms. Within each burst-length subgroup, incremental contrast leakage was seen with mean arterial pressure elevation (ρ=0.2-0.8). CONCLUSIONS: We describe a novel MR imaging-integrated real-time swine intracerebral hemorrhage model of acute hematoma growth and contrast extravasation.

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.

A three-dimensional model of an ultrasound contrast agent gas bubble and its mechanical effects on microvessels.

Ultrasound contrast agents inside a microvessel, when driven by ultrasound, oscillate and induce mechanical stresses on the vessel wall. These mechanical stresses can produce beneficial therapeutic effects but also induce vessel rupture if the stresses are too high. Therefore, it is important to use sufficiently low pressure amplitudes to avoid rupturing the vessels while still inducing the desired therapeutic effects. In this work, we developed a comprehensive three-dimensional model of a confined microbubble inside a vessel while considering the bubble shell properties, blood viscosity, vessel wall curvature and the mechanical properties of the vessel wall. Two bubble models with the assumption of a spherical symmetric bubble and a simple asymmetrical bubble were simulated. This work was validated with previous experimental results and enabled us to evaluate the microbubbles' behaviour and the resulting mechanical stresses induced on the vessel walls. In this study, the fluid shear and circumferential stresses were evaluated as indicators of the mechanical stresses. The effects of acoustical parameters, vessel viscoelasticity and rigidity, vessel/bubble size and off-centre bubbles on bubble behaviour and stresses on the vessel were investigated. The fluid shear and circumferential stresses acting on the vessel varied with time and location. As the frequency changed, the microbubble oscillated with the highest amplitude at its resonance frequency which was different from the resonance frequency of an unbound bubble. The bubble resonance frequency increased as the rigidity of a flexible vessel increased. The fluid shear and circumferential stresses peaked at frequencies above the bubble's resonance frequency. The more rigid the vessels were, the more damped the bubble oscillations. The synergistic effect of acoustic frequency and vessel elasticity had also been investigated since the circumferential stress showed either an increasing trend or a decreasing one versus the vessel rigidity at different acoustic frequencies. When the acoustic pressure was increased from 52 to 680 kPa, the maximum bubble radius increase by 2.5 fold, and the maximum shear and circumferential stress increased by 15.7 and 18.3 fold, respectively. The shear stress was largest when the acoustic frequency was higher (3.25 MHz) and the ratio of the vessel radius to the bubble radius was lower. The circumferential stress was largest when the bubble wall was closer to the vessel wall. An oscillating off-centre bubble forms a mushroom shape with the most damping on the points closest to the vessel wall.

A scanned, focused, multiple transducer ultrasonic system for localized hyperthermia treatments. 1987.

A commercial diagnostic ultrasound scanner (Octoson) was modified for performing hyperthermia treatments. The temperature elevations were induced in tissues by four large, focused ultrasonic transducers whose common focal zone was scanned along a computer controlled path as determined from B-scan images. The system is described and the results of preliminary tests demonstrating some of its capabilities are given. Extensive tests with canine thighs and kidneys were performed. The blood flow to the kidneys was controllable, and thus tumours having different blood perfusion rates could be simulated. The results showed that the system is capable of inducing a local temperature maximum deep in tissues (up to 10 cm was tested) and that tissues with high perfusion rates could be heated.

Computational aspects in high intensity ultrasonic surgery planning.

Therapeutic ultrasound treatment planning is discussed and computational aspects regarding it are reviewed. Nonlinear ultrasound simulations were solved with a combined frequency domain Rayleigh and KZK model. Ultrasonic simulations were combined with thermal simulations and were used to compute heating of muscle tissue in vivo for four different focused ultrasound transducers. The simulations were compared with measurements and good agreement was found for large F-number transducers. However, at F# 1.9 the simulated rate of temperature rise was approximately a factor of 2 higher than the measured ones. The power levels used with the F# 1 transducer were too low to show any nonlinearity. The simulations were used to investigate the importance of nonlinarities generated in the coupling water, and also the importance of including skin in the simulations. Ignoring either of these in the model would lead to larger errors. Most notably, the nonlinearities generated in the water can enhance the focal temperature by more than 100%. The simulations also demonstrated that pulsed high power sonications may provide an opportunity to significantly (up to a factor of 3) reduce the treatment time. In conclusion, nonlinear propagation can play an important role in shaping the energy distribution during a focused ultrasound treatment and it should not be ignored in planning. However, the current simulation methods are accurate only with relatively large F-numbers and better models need to be developed for sharply focused transducers.

Clinical applications of focused ultrasound-the brain.

This paper provides a historic and contemporary overview of the use of focused ultrasound for treating brain disorders.

Longitudinal and shear mode ultrasound propagation in human skull bone.

Recent studies have attempted to dispel the idea of the longitudinal mode being the only significant mode of ultrasound energy transport through the skull bone. The inclusion of shear waves in propagation models has been largely ignored because of an assumption that shear mode conversions from the skull interfaces to the surrounding media rendered the resulting acoustic field insignificant in amplitude and overly distorted. Experimental investigations with isotropic phantom materials and ex vivo human skulls demonstrated that, in certain cases, a shear mode propagation scenario not only can be less distorted, but at times allowed for a substantial (as much as 36% of the longitudinal pressure amplitude) transmission of energy. The phase speed of 1.0-MHz shear mode propagation through ex vivo human skull specimens has been measured to be nearly half of that of the longitudinal mode (shear sound speed = 1500 +/- 140 m/s, longitudinal sound speed = 2820 +/- 40 m/s), demonstrating that a closer match in impedance can be achieved between the skull and surrounding soft tissues with shear mode transmission. By comparing propagation model results with measurements of transcranial ultrasound transmission obtained by a radiation force method, the attenuation coefficient for the longitudinal mode of propagation was determined to between 14 Np/m and 70 Np/m for the frequency range studied, while the same for shear waves was found to be between 94 Np/m and 213 Np/m. This study was performed within the frequency range of 0.2 to 0.9 MHz.

Local frequency dependence in transcranial ultrasound transmission.

The development of large-aperture multiple-source transducer arrays for ultrasound transmission through the human skull has demonstrated the possibility of controlled and substantial acoustic energy delivery into the brain parenchyma without the necessitation of a craniotomy. The individual control of acoustic parameters from each ultrasound source allows for the correction of distortions arising from transmission through the skull bone and also opens up the possibility for electronic steering of the acoustic focus within the brain. In addition, the capability to adjust the frequency of insonation at different locations on the skull can have an effect on ultrasound transmission. To determine the efficacy and applicability of a multiple-frequency approach with such a device, this study examined the frequency dependence of ultrasound transmission in the range of 0.6-1.4 MHz through a series of 17 points on four ex vivo human skulls. Effects beyond those that are characteristic of frequency-dependent attenuation were examined. Using broadband pulses, it was shown that the reflected spectra from the skull revealed information regarding ultrasound transmission at specific frequencies. A multiple-frequency insonation with optimized frequencies over the entirety of five skull specimens was found to yield on average a temporally brief 230% increase in the transmitted intensity with an 88% decrease in time-averaged intensity transmission within the focal volume. This finding demonstrates a potential applicability of a multiple-frequency approach in transcranial ultrasound transmission.

Targeted disruption of the blood-brain barrier with focused ultrasound: association with cavitation activity.

Acoustic emission was monitored during focused ultrasound exposures in conjunction with an ultrasound contrast agent (Optison) in order to determine if cavitation activity is associated with the induction of blood-brain barrier disruption (BBBD). Thirty-four locations were sonicated (frequency: 260 kHz) at targets 10 mm deep in rabbit brain (N = 9). The sonications were applied at peak pressure amplitudes ranging from 0.11 to 0.57 MPa (burst length: 10 ms; repetition frequency of 1 Hz; duration: 20 s). Acoustic emission was recorded with a focused passive cavitation detector. This emission was recorded at each location during sonications with and without Optison. Detectable wideband acoustic emission was observed only at 0.40 and 0.57 MPa. BBBD was observed in contrast MRI after sonication at 0.29-0.57 MPa. The appearance of small regions of extravasated erythrocytes appeared to be associated with this wideband emission signal. The results thus suggest that BBBD resulting from focused ultrasound pulses in the presence of Optison can occur without indicators for inertial cavitation in vivo, wideband emission and extravasation. If inertial cavitation is not responsible for the BBBD, other ultrasound/microbubble interactions are likely the source. A significant increase in the emission signal due to Optison at the second and third harmonics of the ultrasound driving frequency was found to correlate with BBBD and might be useful as an online method to indicate when the disruption occurs.

Resonance frequency of microbubbles in small blood vessels: a numerical study.

Microbubbles are currently used as ultrasound contrast agents. Their potential therapeutic applications are also under investigation. This work is designed to provide some insight into the mechanisms of energy absorption and deposition by a preformed gas bubble in the microvasculature to optimize its efficacy. In the linear regime, the most favourable condition for the transfer of energy from an ultrasonic field to a gas bubble occurs when the centre frequency of the ultrasonic field equals the resonance frequency of the bubble. The resonance frequency of gas microbubbles has been investigated up to now mainly in unbounded liquids; however when bubbles are confined in small regions, their resonance frequency is strongly affected by the surrounding boundaries. A parametric study on how the resonance frequency of microbubbles in blood vessels is affected by the bubble radius, vessel radius and the bubble position in the vessel is presented. The resonance frequency decreases below its free value with decreasing vessel radius for vessels smaller than 200-300 microm depending on the bubble size. This model suggests the possibility of using ultrasound in a range of frequencies that are, in general, lower than the ones used now for therapeutic and diagnostic applications of ultrasound (a few MHz). When microbubbles oscillate at their resonance frequency they absorb and therefore emit more energy. This energy may allow specific blood vessels to be targeted for both diagnostic and therapeutic applications of ultrasound.

Theoretical and experimental validation of a dual-frequency excitation method for spatial control of cavitation.

Inertial cavitation has been implicated as the primary mechanism for a host of emerging applications. In all these applications, the main concern is to induce cavitation in perfectly controlled locations in the field; this means specifically to be able to achieve the cavitation threshold at the geometrical focus of the transducer without stimulating its near field. In this study, we develop dual-frequency methods to preferentially lower the cavitation threshold at the focus relative to the rest of the field. Two families of dual-frequency driving waveforms are evaluated in a bubble model incorporating rectified diffusion. Results are then verified by experiment. Finally, the performance of the rest of acoustic field in suppressing cavitation when cavitation is induced at the focus is investigated theoretically and checked experimentally. This study shows that dual-frequency phased arrays could be used to precisely control cavitation. The cavitation threshold is proved to be 1.2 times higher in the near field than at the focus. The concept of cavitation field is introduced and complements cavitation studies concentrating on the focal behaviour only.

Ultrasound phase-contrast transmission imaging of localized thermal variation and the identification of fat/tissue boundaries.

We present a new ultrasound technique for registering localized temperature changes in soft tissues. Conversely, small temperature changes may be induced in order to image tissue layers. The concept is motivated by the search for a compact, low cost method for guiding noninvasive thermal therapies; however its utility may extend to a wide range of imaging problems such as tumour imaging in the breast. This method combines ultrasound transmission imaging, planar projection techniques and phase-contrast theory. After outlining the theoretical foundation of the technique, its feasibility is tested by simulating localized heating within homogeneous tissue layers. Success of this imaging method is evaluated as a function of the ultrasound-imaging wavelength for a Gaussian-shaped heated region over the frequency range from 0.1 to 2 MHz. Furthermore we simulate two-dimensional image reconstruction from a receiving array. We conclude that thermal phase-contrast imaging in tissues is plausible for detecting the treatment spot in thermal therapies while operating at frequencies below 1 MHz. Additionally, it may also be possible to use the method for noninvasive thermometry. However, thermometry would require operation at higher frequencies at the tradeoff of increased attenuation and higher sensitivity to scattering, which needs to be further explored.

Superresolution ultrasound imaging using back-projected reconstruction.

An ultrasound technique for imaging objects significantly smaller than the source wavelength is investigated. Signals from a focused beam are recorded over an image plane in the acoustic farfield and backprojected in the wave-vector domain to the focal plane. A superresolution image recovery method is then used to analyze the Fourier spatial frequency spectrum of the signal in an attempt to deduce the location and size of objects in this plane. The physical foundation for the method is rooted in the fact that high spatial frequencies introduced by the object in fact affect the lower (nonevanescent) spatial frequencies of the overall signal. The technique achieves this by using a priori measurements of the ultrasound focus in water, which gives full spectral information about the image source. A guess is then made regarding the size and location of the object that distorted the field, and this is convolved with the a priori measurement, thus creating a candidate image. A large number of candidates are generated and the one whose spectrum best matches the uncorrected image is accepted. The method is demonstrated using 0.34- and 0.60-mm wires with a focused 1.05-MHz ultrasound signal and then a human hair (approximately 0.03 mm) with a 4.7-MHz signal.

Forced linear oscillations of microbubbles in blood capillaries.

A theoretical investigation of the forced linear oscillations of a gas microbubble in a blood capillary, whose radius is comparable in size to the bubble radius is presented. The natural frequency of oscillation, the thermal and viscous damping coefficients, the amplitude resonance, the energy resonance, as well as the average energy absorbed by the system, bubble plus vessel, have been computed for different kinds of gas microbubbles, containing air, octafluropropane, and perflurobutane as a function of the bubble radius and applied frequency. It has been found that the bubble behavior is isothermal at low frequencies and for small bubbles and between isothermal and adiabatic for larger bubbles and higher frequencies, with the viscous damping dominating over the thermal damping. Furthermore, the width of the energy resonance is strongly dependent on the bubble size and the natural frequency of oscillation is affected by the presence of the vessel wall and position of the bubble in the vessel. Therefore, the presence of the blood vessel affects the way in which the bubble absorbs energy from the ultrasonic field. The motivation of this study lies in the possibility of using gas microbubbles as an aid to therapeutic focused ultrasound treatments.

Enhanced ultrasound transmission through the human skull using shear mode conversion.

A new transskull propagation technique, which deliberately induces a shear mode in the skull bone, is investigated. Incident waves beyond Snell's critical angle experience a mode conversion from an incident longitudinal wave into a shear wave in the bone layers and then back to a longitudinal wave in the brain. The skull's shear speed provides a better impedance match, less refraction, and less phase alteration than its longitudinal counterpart. Therefore, the idea of utilizing a shear wave for focusing ultrasound in the brain is examined. Demonstrations of the phenomena, and numerical predictions are first studied with plastic phantoms and then using an ex vivo human skull. It is shown that at a frequency of 0.74 MHz the transskull shear method produces an amplitude on the order of--and sometimes higher than--longitudinal propagation. Furthermore, since the shear wave experiences a reduced overall phase shift, this indicates that it is plausible for an existing noninvasive transskull focusing method [Clement, Phys. Med. Biol. 47(8), 1219-1236 (2002)] to be simplified and extended to a larger region in the brain.