Controlled tissue emulsification produced by high intensity focused ultrasound shock waves and millisecond boiling

In high intensity focused ultrasound (HIFU) applications, tissue may be thermally necrosed by heating, emulsified by cavitation, or, as was recently discovered, emulsified using repetitive millisecond boiling caused by shock wave heating. Here, this last approach was further investigated. Experiments were performed in transparent gels and ex vivo bovine heart tissue using 1, 2, and 3 MHz focused transducers and different pulsing schemes in which the pressure, duty factor, and pulse duration were varied. A previously developed derating procedure to determine in situ shock amplitudes and the time-to-boil was refined. Treatments were monitored using B-mode ultrasound. Both inertial cavitation and boiling were observed during exposures, but emulsification occurred only when shocks and boiling were present. Emulsified lesions without thermal denaturation were produced with shock amplitudes sufficient to induce boiling in less than 20 ms, duty factors of less than 0.02, and pulse lengths shorter than 30 ms. Higher duty factors or longer pulses produced varying degrees of thermal denaturation combined with mechanical emulsification. Larger lesions were obtained using lower ultrasound frequencies. The results show that shock wave heating and millisecond boiling is an effective and reliable way to emulsify tissue while monitoring the treatment with ultrasound.     

Effects of adjacent bubble on spatiotemporal evolutions of mechanical stresses surrounding bubbles oscillating in tissues

The cavitation dynamics and mechanical stress in viscoelastic tissues, as the primary mechanisms of some ultrasound therapies, are extremely complex due to the interactions of cavitation bubble with adjacent bubbles and surrounding tissues. Therefore, the cavitation dynamics and resultant mechanical stress of two-interacting bubbles in the viscoelastic tissues are numerically investigated, especially focusing on the effects of the adjacent bubble. The results demonstrate that the mechanical stress is highly dependent on the bubble dynamics. The compressive stress and tensile stress are generated at the stage of bubble expansion and collapse stage, respectively. Furthermore, within the initial parameters examined in this paper, the effects of the adjacent bubble will distinctly suppress the radial expansion of the small bubble and consequently lead its associated stresses to decrease. Owing to the superimposition of two stress fields, the mechanical stresses surrounding the small bubble in the direction of the neighboring bubble are smaller than those in other directions. For two interacting cavitation bubbles, the suppression effects of the nearby bubble on both the cavitation dynamics and the stresses surrounding the small bubble increase as the ultrasound amplitude and the initial radius of the large bubble increase, whereas they decrease with the inter-bubble distance increasing. Moreover, increasing the tissue viscoelasticity will reduce the suppression effects of the nearby bubble, except in instances where the compressive stress and tensile stress first increase and then decrease with the tissue elasticity and viscosity increasing respectively. This study can provide a further understanding of the mechanisms of cavitation-associated mechanical damage to the adjacent tissues or cells.     

Bubble dynamics and size distributions during focused ultrasound insonation

The deposition of ultrasonic energy in tissue can cause tissue damage due to local heating. For pressures above a critical threshold, cavitation will occur, inducing a much larger thermal energy deposition in a local region. The present work develops a nonlinear bubble dynamics model to numerically investigate bubble oscillations and bubble-enhanced heating during focused ultrasound (HIFU) insonation. The model is applied to calculate two threshold-dependent phenomena occurring for nonlinearly oscillating bubbles: Shape instability and growth by rectified diffusion. These instabilities in turn are shown to place physical boundaries on the time-dependent bubble size distribution, and thus the thermal energy deposition.     

超声造影剂微泡非线性动力学响应的机理及相关应用

随着生命科学及现代医学的发展, 一体化无创精准诊疗已经日益成为人们关注的焦点问题, 而关于超声造影剂微泡的非线性效应的相关机理、动力学建模及其在超声医学领域中的应用研究也得到了极大的推动. 本文对下列课题进行了总结和讨论, 包括: 1)基于Mie散射技术和流式细胞仪对造影剂微泡参数进行定征的一体化解决方案; 2)通过对微泡包膜的黏弹特性进行非线性修正, 构建新的包膜微泡动力学模型; 3)探索造影剂惯性空化阈值与其包膜参数之间的相关性; 以及4)研究超声联合造影剂微泡促进基因/药物转染效率并有效降低其生物毒性的相关机理.     

Effect of temperature on rectified diffusion during ultrasound-induced heating

Experimental observations of delayed-onset cavitation during ultrasound insonation have been suggested as being caused by a change in the size distribution of the bubble population due to rectified diffusion. To investigate this hypothesis, a single bubble model is used here to explore the effect of heating and the subsequent elevated temperatures on the rectified diffusion process. Numerical solution of the model, which includes the temperature dependences of seven relevant physical parameters, allows quantification of the change in the pressure threshold for rectified diffusion, as well as the importance of the bulk liquid saturation concentration in determining bubble evolution. Although elevated temperatures and liquid supersaturation reduce the rectified diffusion threshold, it remains coincident with the inertial cavitation thresholds at submicron bubble sizes at all temperatures. This observation suggests that changes in the nucleation environment, rather than bubble growth due to rectified diffusion, is a more likely cause of delayed-onset cavitation events.     

Passive cavitation mapping for localization and tracking of bubble dynamics

Current acoustic techniques for studying cavitation dynamics are only readily applicable to single-bubble activity, while optical methods can only be used in transparent media. However, multi-bubble cavitation often occurs in opaque media such as biological tissue. Here, the signals received passively by each of the 64 channels of a diagnostic ultrasound array are used to localize and separate emissions from several bubble clusters cavitating in agar gel, thereby providing a method of observing cavitation dynamics. The method has a high spatiotemporal resolution and is applicable to cavitation in opaque media.     

Estimation of temperature elevation generated by ultrasonic irradiation in biological tissues using the thermal wave method

In most previous models,simulation of the temperature generation in tissue is based on the Pennes bio-heat transfer equation,which implies an instantaneous thermal energy deposition in the medium.Due to the long thermal relaxation time τ(20 s-30 s) in biological tissues,the actual temperature elevation during clinical treatments could be different from the value predicted by the Pennes bioheat equation.The thermal wave model of bio-heat transfer(TWMBT) defines a thermal relaxation time to describe the tissue heating from ultrasound exposure.In this paper,COMSOL Multiphysics 3.5a,a finite element method software package,is used to simulate the temperature response in tissues based on Pennes and TWMBT equations.We further discuss different factors in the bio-heat transfer model on the influence of the temperature rising and it is found that the temperature response in tissue under ultrasound exposure is a rising process with a declining rate.The thermal relaxation time inhibits the temperature elevation at the beginning of ultrasonic heating.Besides,thermal relaxation in TWMBT leads to lower temperature estimation than that based on Pennes equation during the same period of time.The blood flow carrying heat dominates most to the decline of temperature rising rate and the influence increases with temperature rising.On the contrary,heat diffusion,which can be described by thermal conductivity,has little effect on the temperature rising.     

The potential for thermal damage posed by microbubble ultrasound contrast agents

The development of coated microbubble ultrasound contrast agents for use in imaging applications and as carriers in drug and gene delivery applications has intensified the need for a clear understanding of their behaviour and potential bioeffects. Previous studies have focused on the risks posed by unencapsulated bubbles as representing the "worst case scenario". They have concluded that the risk of thermal damage should be minimal provided the threshold for inertial cavitation is not exceeded. However, these treatments have ignored the heating effects due to viscous dissipation in the coatings of contrast agent particles. Simulations indicate that the temperature rise due to this process may be sufficient to generate harmful bioeffects even under conventionally "safe" insonation conditions. The implications of these findings and strategies for addressing the risks posed by contrast agents are discussed.     

Numerical investigation on acoustic cavitation characteristics of an air-vapor bubble: Effect of equation of state for interior gases

The cavitation dynamics of an air-vapor mixture bubble with ultrasonic excitation can be greatly affected by the equation of state (EOS) for the interior gases. To simulate the cavitation dynamics, the Gilmore-Akulichev equation was coupled with the Peng–Robinson (PR) EOS or the Van der Waals (vdW) EOS. In this study, the thermodynamic properties of air and water vapor predicted by the PR and vdW EOS were first compared, and the results showed that the PR EOS gives a more accurate estimation of the gases within the bubble due to the less deviation from the experimental values. Moreover, the acoustic cavitation characteristics predicted by the Gilmore-PR model were compared to the Gilmore-vdW model, including the bubble collapse strength, the temperature, pressure and number of water molecules within the bubble. The results indicated that a stronger bubble collapse was predicted by the Gilmore-PR model rather than the Gilmore-vdW model, with higher temperature and pressure, as well as more water molecules within the collapsing bubble. More importantly, it was found that the differences between both models increase at higher ultrasound amplitudes or lower ultrasound frequencies while decreasing as the initial bubble radius and the liquid parameters (e.g., surface tension, viscosity and temperature of the surrounding liquid) increase. This study might offer important insights into the effects of the EOS for interior gases on the cavitation bubble dynamics and the resultant acoustic cavitation-associated effects, contributing to further optimization of its applications in sonochemistry and biomedicine.     

Laser-enhanced thermal effects during focused ultrasound radiation on biological tissues

The thermal effects induced by a moderate intensity focused ultrasound and enhanced by combined laser pulses for bio-tissues and tissue-phantom are studied experimentally and theoretically. At first, the heating effects of bio-tissues and tissue-phantom induced by ultrasound and enhanced by laser are measured experimentally. The heating processes induced by attenuations of focused ultrasonic waves and cavitation effects of the focused ultrasound and combined laser are analyzed theoretically. By analyzing the mechanisms of these effects, it is found that the laser nucleation makes the cavitation bubble generation more easily, which can effectively enhance the ultrasonic cavitation effects, and then enhance the thermal effects of the samples. On the other hand, to evaluate quantitatively the heating processes induced by the focused ultrasound and enhanced by the pulsed laser, by fitting the theoretical calculations to the experimental results, the corresponding cavitation bubbles and rising temperatures induced by the focused ultrasound with and without laser can be estimated approximately.     

激光协同聚焦超声辐照生物样品增强热效应的实验和理论研究

对中等强度聚焦超声在生物样品中产生的热效应以及激光协同超声增强热效应进行了实验和理论研究。实验上,对生物和仿生样品在超声作用和激光协同超声作用下加热情况进行测量,通过对比表明,激光协同超声作用于生物样品,引起空化效应以及温度升高更为明显。同时,理论上对聚焦超声在生物样品中衰减产生的热效应、超声空化以及激光协同超声增强空化及其产生热效应进行机理分析。通过对机理的分析表明,激光引起的光致核化使超声空化更易于产生,有效的增强空化效应,进而增强热效应。为对具体实验给出量化分析和估算,通过理论与实验结果相拟合,对超声传播引起的温度升高进行计算,并估算超声和激光协同超声产生空化微泡对加热效应的不同贡献,为空化效应在超声治疗中的贡献提供参考数据。      

A multi-gate time-of-flight technique for estimation of temperature distribution in heated tissue: theory and computer simulation

Non-invasive determination of temperature distribution in biological media is important in many heating-related studies, such as thermal treatment. In this paper, we present an in vitro ultrasound technique for estimation of temperature distribution in heated tissue. Our technique consists of two major steps: (1) using multiple time gates to track echo signals scattered from tissue regions at different depths; (2) estimating temperature distribution based on heating-induced changes of arrival times of echo signals scattered from the targeted tissue regions. We use the conventional cross-correlation approach to track echoes. For temperature estimation, we have developed an iterative method that takes into account the influences of thermal expansion and heating-induced change in the speed of sound on the time of flight. We have introduced a concept of thermal sensitivity of the time of flight and used it to derive a theoretical formula that relates the achievable accuracy on the estimation of tissue temperature to seven parameters. The seven parameters are tissue thermal sensitivity of the time of flight, signal-to-noise ratio, bandwidth and center frequency of the signal, degree of signal decorrelation induced by changes in tissue physical properties during tissue heating, and widths and spacing of the time gates. We tested our technique by computer simulation, using a random discrete scatterer model and temperature distribution data acquired in our laser heating experiments on prostate tissue of live dog. Simulation results showed that our technique could accurately estimate the temperature distribution in the heated tissue. Our technique is fast in terms of computation and could be used as a research tool for in vitro real-time monitoring of temperature distribution in tissue under hyperthermal heating.     

A reduced-order, single-bubble cavitation model with applications to therapeutic ultrasound

Cavitation often occurs in therapeutic applications of medical ultrasound such as shock-wave lithotripsy (SWL) and high-intensity focused ultrasound (HIFU). Because cavitation bubbles can affect an intended treatment, it is important to understand the dynamics of bubbles in this context. The relevant context includes very high acoustic pressures and frequencies as well as elevated temperatures. Relative to much of the prior research on cavitation and bubble dynamics, such conditions are unique. To address the relevant physics, a reduced-order model of a single, spherical bubble is proposed that incorporates phase change at the liquid-gas interface as well as heat and mass transport in both phases. Based on the energy lost during the inertial collapse and rebound of a millimeter-sized bubble, experimental observations were used to tune and test model predictions. In addition, benchmarks from the published literature were used to assess various aspects of model performance. Benchmark comparisons demonstrate that the model captures the basic physics of phase change and diffusive transport, while it is quantitatively sensitive to specific model assumptions and implementation details. Given its performance and numerical stability, the model can be used to explore bubble behaviors across a broad parameter space relevant to therapeutic ultrasound.     

Effects of dissolved gases and an echo contrast agent on ultrasound mediated in vitro gene transfection

The effects of acoustic cavitation on in vitro transfection by ultrasound were investigated. HeLa cells were exposed to 1.0 MHz continuous ultrasound in culture media containing the luciferase gene. Transfection efficiency was elevated when an echo contrast agent, Levovist was added or air was dissolved in the medium. When cells were sonicated in medium saturated with Ar, N2 or N2O which have different gamma values (Cp/Cv), or were saturated with He, Ar or Ne with different thermal conductivities, the effectiveness for the dissolved gases in the ultrasound mediated transfection was Ar > N2 > N2O or Ar > Ne > He, respectively. When free radical formation in water by ultrasound was monitored as a measure of inertial cavitation, it was similarly affected by dissolved gases. These results indicate that the efficiency of ultrasound mediated transfection was significantly affected either by occurrence of or by modification of inertial cavitation due to various gases.     

Effect of acoustic nonlinearity on heating of biological tissue by high-intensity focused ultrasound

Effect of strong acoustic nonlinearity on the efficiency of heating of a biological tissue by high-intensity focused ultrasound in the modes of operation used in real clinical setups is studied. The spatial distributions of thermal sources and the corresponding temperature increments caused by ultrasonic absorption are analyzed. Numerical algorithms are developed for simulating the nonlinear focusing of ultrasound in the calculations of both the heat sources on the basis of the Khokhlov-Zabolotskaya-Kuznetsov-type equations and the temperature field in a tissue on the basis of an inhomogeneous thermal conduction equation with a relaxation term. It is demonstrated that in the mode of operation typical of acoustic surgery, the nonlinearity improves the locality of heating and leads to an increase in the power of thermal sources in the focus by approximately an order of magnitude. The diffusion phenomena in the tissue lead to a smoothing of the spatial temperature distributions, as compared to the distributions of thermal sources. In the case of one-second exposure in the nonlinear mode of focusing, the maximal temperature in the focus exceeds the values obtained in the approximation of linear wave propagation by a factor of three.     

Sonochemiluminescence observation and acoustic detection of cavitation induced by pulsed HIFU at a tissue–fluid interface

The aim of this study is to investigate the mechanism of the erosion process induced by 1.2 MHz pulsed high-intensity focused ultrasound (pulsed HIFU). By using Sonochemiluminescence (SCL) photograph, the initiation and maintenance of active cavitation were observed. In order to understand the role of both inertial cavitation and stable cavitation, a passive cavitation detection (PCD) transducer was used. Since the exposure variables of HIFU are important in the controlled ultrasound tissue erosion, the influence of pulse length (PL) and duty cycle (DC, T_on:T_off) has been examined. The results of tissue hole, SCL observation and acoustic detection revealed that the erosion was highly efficient for shorter PL. For higher DCs, the area of SCL increased with increasing PL. For lower DCs, the area of SCL increased with increasing PL from 10 to 20 μs and then kept constant. For all PLs, the intensity of SCL decreased with lower DC. For all DCs, the intensity of SCL per unit area (the ratio of SCL intensity to SCL area) also decreased with increasing PL from 10 to 80 μs, which suggested that the higher the intensity of SCL is, the higher the efficiency of tissue erosion is. At DC of 1:10, the position of the maximum pixel in SCL pictures was distant from the tissue–fluid interface with the increasing PL because of shielding effect. By the comparison of inertial cavitation dose (ICD) and the stable cavitation dose (SCD), the mechanisms associated with inertial cavitation are very likely to be the key factor of the erosion process.     

Efficient and controllable thermal ablation induced by short-pulsed HIFU sequence assisted with perfluorohexane nanodroplets

A HIFU sequence with extremely short pulse duration and high pulse repetition frequency can achieve thermal ablation at a low acoustic power using inertial cavitation. Because of its cavitation-dependent property, the therapeutic outcome is unreliable when the treatment zone lacks cavitation nuclei. To overcome this intrinsic limitation, we introduced perfluorocarbon nanodroplets as extra cavitation nuclei into short-pulsed HIFU-mediated thermal ablation. Two types of nanodroplets were used with perfluorohexane (PFH) as the core material coated with bovine serum albumin (BSA) or an anionic fluorosurfactant (FS) to demonstrate the feasibility of this study. The thermal ablation process was recorded by high-speed photography. The inertial cavitation activity during the ablation was revealed by sonoluminescence (SL). The high-speed photography results show that the thermal ablation volume increased by ∼643% and 596% with BSA-PFH and FS-PFH, respectively, than the short-pulsed HIFU alone at an acoustic power of 19.5 W. Using nanodroplets, much larger ablation volumes were created even at a much lower acoustic power. Meanwhile, the treatment time for ablating a desired volume significantly reduced in the presence of nanodroplets. Moreover, by adjusting the treatment time, lesion migration towards the HIFU transducer could also be avoided. The SL results show that the thermal lesion shape was significantly dependent on the inertial cavitation in this short-pulsed HIFU-mediated thermal ablation. The inertial cavitation activity became more predictable by using nanodroplets. Therefore, the introduction of PFH nanodroplets as extra cavitation nuclei made the short-pulsed HIFU thermal ablation more efficient by increasing the ablation volume and speed, and more controllable by reducing the acoustic power and preventing lesion migration.     

Passive imaging with pulsed ultrasound insonations

Previously, passive cavitation imaging has been described in the context of continuous-wave high-intensity focused ultrasound thermal ablation. However, the technique has potential use as a feedback mechanism for pulsed-wave therapies, such as ultrasound-mediated drug delivery. In this paper, results of experiments and simulations are reported to demonstrate the feasibility of passive cavitation imaging using pulsed ultrasound insonations and how the images depend on pulsed ultrasound parameters. The passive cavitation images were formed from channel data that was beamformed in the frequency domain. Experiments were performed in an invitro flow phantom with an experimental echo contrast agent, echogenic liposomes, as cavitation nuclei. It was found that the pulse duration and envelope have minimal impact on the image resolution achieved. The passive cavitation image amplitude scales linearly with the cavitation emission energy. Cavitation images for both stable and inertial cavitation can be obtained from the same received data set.     

Modeling cavitation nucleation from laser-illuminated nanoparticles subjected to acoustic stress

In an earlier work by Farny et al. [ARLO 6, 138-143 (2005).] it was demonstrated that the acoustic cavitation threshold in a tissue mimicking gel phantom can be lowered from 4.5 to ～1 MPa by "seeding" the optically transparent phantom with light absorptive gold nanoparticles and irradiating these absorbers with nanosecond pulses of laser light at intensities less than 10 mJ/cm(2). As a follow-up study, a three-stage numerical model was developed to account for prenucleation heating, the nucleation and formation of the vapor cavity, and the resulting vapor bubble dynamics. Through examination of the radius-time evolution of the cavity, the combined thresholds for laser radiant exposure and acoustic peak pressure required to induce inertial cavitation are deduced. It is found that the threshold pressure decreases when laser exposure increases; but the rate depends on exposure levels and the size of the particle. Investigations of the roles of particle size and laser pulse length are performed and optimum choices for these parameters determined in order to obtain inertial cavitation at the lowest possible acoustic pressure and laser intensity.     

Promoting inertial cavitation by nonlinear frequency mixing in a bifrequency focused ultrasound beam

Enhancing cavitation activity with minimal acoustic intensities could be interesting in a variety of therapeutic applications where mechanical effects of cavitation are needed with minimal heating of surrounding tissues. The present work focuses on the relative efficiency of a signal combining two neighbouring frequencies and a one-frequency signal for initiating ultrasound inertial cavitation. Experiments were carried out in a water tank, using a 550 kHz piezoelectric composite spherical transducer focused on targets with 46 μm roughness. The acoustic signal scattered, either by the target or by the cavitation bubbles, is filtered using a spectral and cepstral-like method to obtain an inertial cavitation activity measurement. The ultrasound excitations consist of 1.8 ms single bursts of single frequency f₀ = 550 kHz excitation, in the monofrequency case, and of dual frequency f₁ = 535 kHz and f₂ = 565 kHz excitation, in the bifrequency case. It is shown that depending on the value of the monofrequency cavitation threshold intensity the bifrequency excitation can increase or reduce the cavitation threshold. The analysis of the thresholds indicates that the mechanisms involved are nonlinear. The progress of the cavitation activity beyond the cavitation threshold is also studied. The slope of the cavitation activity considered as a function of the acoustic intensity is always steeper in the case of the bifrequency excitation. This means that the delimitation of the region where cavitation occurs should be cleaner than with a classical monofrequency excitation.