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.     

纳米包膜造影微泡的小波检测技术研究

用小波变换的方法检测人体组织的微小血流灌注包膜造影微泡回波。结合超声造影剂微泡振动的物理模型,构造了不同外加声场条件下的气泡小波,对原始信号进行小波变换,将变换后得到的小波系数分离;进而从微小血流灌注组织杂波中检测出造影剂微泡回波信号。心肌条件下的计算机模拟以及对灌注组织的仿体实验结果表明:与普通母小波相比,基于气泡振动模型的小波,因为是通过理论模型构造所得,已先验表征了造影剂微泡在声场中的回波特性,因此与实验中产生的回波信号具有更为紧密的相关性,从而经小波变换后,造影剂微泡回波信号与组织杂波产生了更高强度的信噪比,及更明显的可分离和对比效果。同时,构造了一个完备的母小波函数库,进一步提高了本方法的适用性以及健壮性。     

Detection procedures of ultrasound contrast agents

In the early days, it was believed that ultrasound contrast agents (UCA) could be sufficiently detected and imaged with the conventional imaging methods nowadays referred to as fundamental imaging. Newer imaging techniques proved to be more sensitive and are based on specific properties of the UCA. In general, these new characteristics involve non-linear and transient characteristics of contrast agents that appear at the high end of the diagnostic acoustic intensity. Imaging modalities used today for UCA are, besides fundamental imaging, second harmonic imaging, power Doppler, harmonic power Doppler, pulse inversion and pulse inversion Doppler, multi-pulse imaging and subharmonic imaging. Although the results of conventional second harmonic imaging are still not optimal for perfusion imaging applications, in combination with Doppler techniques (colour Doppler, power Doppler) it is one of the most sensitive techniques currently available in terms of agent-to-tissue ratio. Further improvements in current and future detection methods demand a complete understanding of the ultrasound-UCA interaction.     

Microbubble spectroscopy of ultrasound contrast agents

A new optical characterization of the behavior of single ultrasound contrast bubbles is presented. The method consists of insonifying individual bubbles several times successively sweeping the applied frequency, and to record movies of the bubble response up to 25 million frames/s with an ultrahigh speed camera operated in a segmented mode. The method, termed microbubble spectroscopy, enables to reconstruct a resonance curve in a single run. The data is analyzed through a linearized model for coated bubbles. The results confirm the significant influence of the shell on the bubble dynamics: shell elasticity increases the resonance frequency by about 50%, and shell viscosity is responsible for about 70% of the total damping. The obtained value for shell elasticity is in quantative agreement with previously reported values. The shell viscosity increases significantly with the radius, revealing a new nonlinear behavior of the phospholipid coating.     

Direct observations of ultrasound microbubble contrast agent interaction with the microvessel wall

Many thousands of contrast ultrasound studies have been conducted in clinics around the world. In addition, the microbubbles employed in these examinations are being widely investigated to deliver drugs and genes. Here, for the first time, the oscillation of these microbubbles in small vessels is directly observed and shown to be substantially different than that predicted by previous models and imaged within large fluid volumes. Using pulsed ultrasound with a center frequency of 1 MHz and peak rarefactional pressure of 0.8 or 2.0 MPa, microbubble expansion was significantly reduced when microbubbles were constrained within small vessels in the rat cecum (p<0.05). A model for microbubble oscillation within compliant vessels is presented that accurately predicts oscillation and vessel displacement within small vessels. As a result of the decreased oscillation in small vessels, a large resting microbubble diameter resulting from agent fusion or a high mechanical index was required to bring the agent shell into contact with the endothelium. Also, contact with the endothelium was observed during asymmetrical collapse, not during expansion. These results will be used to improve the design of drug delivery techniques using microbubbles.     

Acoustic behaviour of current ultrasound contrast agents

A general law gives the approximate change in signal level obtained in a particular imaging mode when a suitable contrast agent is added. It also shows that reduction of background signals is essential to overcome limitations found mainly in conventional (linear) ultrasound contrast imaging. Contrast agents contain stabilized microbubbles with very helpful non-linear properties. Acoustic methods for non-destructive and destructive testing of microbubbles are briefly discussed. In the main part, the linear and non-linear acoustic behaviour of various types of contrast agent are described. The latter is useful for new applications in diagnostic ultrasound.     

Stability analysis of ultrasound thick-shell contrast agents

The stability of thick shell encapsulated bubbles is studied analytically. 3-D small perturbations are introduced to the spherical oscillations of a contrast agent bubble in response to a sinusoidal acoustic field with different amplitudes of excitation. The equations of the perturbation amplitudes are derived using asymptotic expansions and linear stability analysis is then applied to the resulting differential equations. The stability of the encapsulated microbubbles to nonspherical small perturbations is examined by solving an eigenvalue problem. The approach then identifies the fastest growing perturbations which could lead to the breakup of the encapsulated microbubble or contrast agent.     

Evaluation of synthetic phospholipid ultrasound contrast agents

The echogenic properties of synthetic, phospholipid encapsulated, air-filled microbubbles with various carbon-chain length as ultrasound contrast agents are investigated through the use of a flow-through laboratory ultrasound system. Specifically, we investigate the effect of shell carbon-chain length on the ultrasonic signal for a variety of flow rates. Averaged, integrated backscatter power measurements from the lipid encapsulated agents are benchmarked against those of Albunex (Albunex is a registered trademark of Molecular Biosystems, Inc., San Diego, CA), a commercially available, air-filled protein microbubbles contrast agent, approved for clinical use in echocardiography in the United States by the Food and Drug Administration. We find that the lipid encapsulated agents sustain less damage leading to gas dissolution or particle destruction as compared to Albunex in the slow-flow studies performed. The carbon-chain length of the encapsulating lipid molecule is shown not to observably affect the backscattered amplitude of ultrasound at flow velocities exceeding 7 mm/s.     

Superhydrophobic silica nanoparticles as ultrasound contrast agents

Microbubbles have been widely studied as ultrasound contrast agents for diagnosis and as drug/gene carriers for therapy. However, their size and stability (lifetime of 5–12 min) limited their applications. The development of stable nanoscale ultrasound contrast agents would therefore benefit both. Generating bubbles persistently in situ would be one of the promising solutions to the problem of short lifetime. We hypothesized that bubbles could be generated in situ by providing stable air nuclei since it has been found that the interfacial nanobubbles on a hydrophobic surface have a much longer lifetime (orders of days). Mesoporous silica nanoparticles (MSNs) with large surface areas and different levels of hydrophobicity were prepared to test our hypothesis. It is clear that the superhydrophobic and porous nanoparticles exhibited a significant and strong contrast intensity compared with other nanoparticles. The bubbles generated from superhydrophobic nanoparticles sustained for at least 30 min at a MI of 1.0, while lipid microbubble lasted for about 5 min at the same settings. In summary MSNs have been transformed into reliable bubble precursors by making simple superhydrophobic modification, and made into a promising contrast agent with the potentials to serve as theranostic agents that are sensitive to ultrasound stimulation.     

Principles and recent developments in ultrasound contrast agents

The behaviour of gas bubbles and gas encapsulated spheres as echographic contrast agents is reviewed. Compared with rigid spheres, gas bubbles are superior scattering agents and they offer a number of useful properties which can be exploited in a variety of ways. The analysis of their velocity of sound, back-scatter intensity, second harmonic emission and resonant frequency opens up new perspectives in the development of contrast agents for echocardiographic research with potential clinical applications.     

超声造影剂研究进展

本文综述超声造影剂的现状及新进展，在简要介绍造影剂物理原理的基础上，对超声造影剂在医学超声领域尤其是低速血流测量与组织灌注成像中的应用进行了总结与讨论，并介绍了用于实验研究和第二期临床试验的国际上最新的超声造影剂的特点，制备技术与评价方法。     

Excitation of polymer-shelled contrast agents with high-frequency ultrasound

Few experimental and complementary theoretical studies have investigated high-frequency (>20 MHz) nonlinear responses from polymer-shelled ultrasound contrast agents. Three polymer agents with different shell properties were examined for their single-bubble backscatter when excited with a 40 MHz tone burst. Higher-order harmonic responses were observed for the three agents; however, their occurrence was at least partly due to nonlinear propagation. Only one of the agents (1.1 microm mean diameter) showed a subharmonic response for longer excitations (approximately 10-15 cycles) and midlevel pressure excitations ( 2.5 MPa). Theoretical calculations of the backscattered spectrum revealed behavior similar to the experimental results in specific parameter regimes.     

Experimental study of the preparation of targeted microbubble contrast agents carrying urokinase and RGDS

Purpose

To explore the feasibility and the best combined proportion of the preparation of targeted and thrombolytic contrast agents carrying urokinase (UK) and Arg-Gly-Asp-Ser (RGDS), and to study its effect of thrombolysis in vitro.

Methods

Urokinase and RGDS were combined to the surface of SonoVue by direct conjugation method. According to the different ratio of urokinase and RGDS when mixed, they were divided into three groups whose urokinase/RGDS were 1:1, 2:1 and 1:2, respectively. To measure the binding rate of microbubbles and urokinase as well as RGDS by flow cytometry. To detect the effect of thrombolysis by thrombolytic experiment in vitro. To detect the targeting effects by experiment in vivo.

Results

The results of flow cytometry detection showed that the binding rates of urokinase and RGDS of three groups were, respectively, 73.4 ± 11.0% and 67.1 ± 10.9%, 8.8 ± 7.2% and 7.8 ± 6.9%, 49.7 ± 21.3% and 45.9 ± 21.7% after standing for 2 h. In vitro thrombolysis experiment indicated that the urokinase had activity in the prepared contrast agent which the binding rates of urokinase and RGDS were the highest. And it was injected intravenous, the contrast agent aggregated on the surface of the thrombus of the rabbit femoral arterial. The thrombus emitted fluorescence.

Conclusions

The binding rate of the targeted contrast agent prepared by 1:1 of urokinase/RGDS was the highest. It had thrombolysis ability in vitro, and it had thrombo-targeting effect in vivo.     

Physical phenomena affecting quantitative imaging of ultrasound contrast agents

Microbubbles stabilized by a surfactant or polymer coating are the most effective form of contrast agent available for ultrasound imaging. They have shown great potential as a means of quantifying tissue perfusion, in particular determining physiologically significant parameters such as relative vascular volume and flow velocity. Clinical implementation of quantitative imaging procedures, however, has been hindered by poor characterisation of the complex relationship between microbubble concentration, scattering and image intensity. The aim of this paper is to describe theoretical and experimental investigations of the physical phenomena underlying these effects, such as the time, pressure and frequency dependence of microbubble behaviour, the influence of the bubble coating, size distribution and concentration; and to discuss the challenges involved in developing accurate quantitative imaging protocols.     

Experimental investigation of the pulse inversion technique for imaging ultrasound contrast agents

The application of ultrasound contrast agents aims to detect low velocity blood flow in the microcirculation. To enhance discrimination between tissue and blood containing the contrast agent, harmonic imaging is used. Harmonic imaging requires the application of narrow-band signals and is obscured by high levels of native harmonics generated in an intervening medium. To improve discrimination between contrast agent and native harmonics, a pulse inversion technique has been proposed. Pulse inversion allows wide-band signals, thus preserving the axial resolution. The present study examines the interference of native harmonics and discusses the practical difficulties of wide-band pulse inversion measurements of harmonics by a single transducer. Native harmonics are not eliminated by pulse inversion. Furthermore, only even harmonics remain and are amplified by 6 dB, alleviating the requirement for selective filtering. Finally, it is shown that the contaminating third harmonic contained in the square wave activation signal leaks through in the emitted signal. The spectral location of the contaminating third harmonic is governed by the transducer spectral characteristics while the location of the native and contrast agent second harmonics is not. Thus the contaminating third harmonic and the native and contrast agent second harmonics may overlap and interfere. Optimal discrimination requires a balance between maximal sensitivity for the second harmonic at reception and minimal interference from the contaminating third harmonic.     

A theoretical investigation of chirp insonification of ultrasound contrast agents

A theoretical investigation of second harmonic imaging of an Ultrasound Contrast Agent (UCA) under chirp insonification is considered. By solving the UCA’s dynamical equation analytically, the effect that the chirp signal parameters and the UCA shell parameters have on the amplitude of the second harmonic frequency are examined. This allows optimal parameter values to be identified which maximise the UCA’s second harmonic response. A relationship is found for the chirp parameters which ensures that a signal can be designed to resonate a UCA for a given set of shell parameters. It is also shown that the shell thickness, shell viscosity and shell elasticity parameter should be as small as realistically possible in order to maximise the second harmonic amplitude. Keller-Herring, Second Harmonic, Chirp, Ultrasound Contrast Agent.     

Study on the multiple scattering effects of ultrasound contrast agents

In this paper, the multiple scattering of interacting encapsulated microbubbles in suspensions is calculated using two novel approaches--Kargl's effective medium approach and Ye and Ding's approach of 2nd-order correction. Two types of contrast agents with bubbles of different sizes and concentrations are chosen for our numerical simulations. One is Albunex, which is depicted by Church and has a size range of several microns, and the other is sodium laureate solution (before fractionation) described by Soetanto and Chan and has an average size of 35.5 microm. The numerical results from these two approaches are compared with that from the linear approximation. It is found that the multiple scattering effects on attenuation and dispersion of sound in suspensions are evident in the cases of high bubble volume fractions, basically greater than the order of 1 x 10(-4), and much more obvious for larger bubbles with average size of tens of microns.     

Isointense model for the evaluation of tumor-specific MRI contrast agents

An isointense model has been developed to evaluate the applicability of putative tumor-specific MRI contrast agents. Data for tissue relaxation measurements in the presence of Mn(III)TPPS4 are used to illustrate the model. The concentration of contrast agent in tumor tissue required for a tumor/normal tissue signal difference-to-noise ratio of 5 (delta SNR = 5) is determined for a T1 weighted pulse sequence and several hypothetical tumor/normal tissue pairs. The impact of various contrast agent characteristics including initial tumor/normal tissue relaxation values, differential uptake of contrast agent, and in vivo relaxivity are considered. Isointense tumor/normal tissue with longer initial relaxation times are shown to be more affected by the presence of contrast agent. In addition those with initially longer relaxation times have less rigorous requirements for tumor specificity. Typically, a normal tissue/tumor uptake ratio of 1:2 increases the concentration required for delta SNR = 5 by a factor of two compared to that of exclusive uptake in tumor. For the T1 weighted pulse sequence employed, the concentration required for delta SNR = 5 is shown to be linear with the inverse of in vivo relaxivity for the hypothetical tissues considered. The isointense model is also extended to predict the field dependence of tumor-specific contrast enhancement by Mn(III)TPPS4.     

Dynamical model of encapsulated gas microbubble under ultrasound based on elastic mechanics

Based on the theory of elastic mechanics, a dynamical model of an encapsulated gas microbubble under ultrasound is presented. The dynamical motion of the microbubble is divided into three states: buckled, elastic, and ruptured. The model describes the compression-only behavior appropriately and derives the transient variation of the resonance frequency of the damped oscillation and the relation between the critical rupture radius and initial outer radius. The normal stress in the tangential direction plays the principal role in the rupture and buckling of the encapsulating shell, resulting in likely rupture for a larger microbubble and resistance to rupture for a thicker-shell microbubble. Comparison of proposed dynamical model with Marmottant’s model has been given. The dynamical model can be employed in ultrasound medical diagnostics and therapy of drug incorporation or extravasation through further understanding the influence of the encapsulating shell.     

准高斯声束对超声造影剂的声辐射力研究*

超声造影剂的定向输运在超声医学成像领域有着极为重要的意义,而声辐射力作用是实现该过程的关键,相比于高斯声束,准高斯声束是无源亥姆霍兹方程的精确解,可以使用标准波分解法简化计算。因此,本文研究了准高斯声束对超声造影剂的声辐射力作用。文章首先分析了准高斯声束与高斯声束之间的相关性;随后通过数值计算求得了准高斯声束对超声造影剂模型的声辐射力函数与无量纲频率之间的关系;最后,本文研究了不同造影剂气泡情况下的声辐射力。研究结果表明:声辐射力函数随无量纲频率变化将在不同位置出现共振峰,不同的波束宽度值将改变辐射力强度,但不改变共振峰的位置。相关结果可为利用声辐射力定向输运超声造影剂至靶向位置提供理论参考。