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.     

Acoustic properties of organic powders as ultrasonic contrast agents

The results of experiments on measuring attenuation and the effective acoustic nonlinear parameter of the second order are given for a suspension of cocoa-powder in water at different concentrations of the suspension. In the process of evaluating the value of the nonlinear parameter the attenuation in the suspension and generation of the second harmonic not only in the suspension but also in water are taken into account. The obtained results are evidence of the possibility of using a suspension of cocoa-powder in water as a technical substitute for ultrasonic contrast agents. The values of attenuation (up to 60 m⁻¹ at the concentration of 1 g of the powder per 1 l of water) and the nonlinear parameter (up to 120 m⁻¹ at the same concentration) mean that the suspension of cocoa-powder in water has smaller attenuation and the nonlinear parameter than ultrasonic contrast agents at the same concentration. However, these values for the suspension differ considerably from corresponding values for water or blood and, therefore, a suspension of cocoa-powder in water is a promising “substitute” for ultrasonic contrast agents in the case of technical testing of systems for nonlinear tomography of a blood flow, but cannot replace them in medical studies.     

Sub-harmonic response from polymer-shelled contrast agents with a 40-MHz excitation

There is a growing interest in using acoustic contrast agents with high-frequency ultrasound (> 15 MHz) in order to better visualize microcirculation. Experiments were performed with polycaprolactone-shelled agents (POINT Biomedical, San Carlos, CA) having mean diameters of 0.56, 1.1, and 3.4 mum. The agents were heavily diluted in filtered water and injected through a 200 mum channel into the focal zone of a 40-MHz transducer that had a focal length of 12 mm and an outer diameter of 6 mm. Backscatter signals from single agents were digitized using tone bursts of 5 to 20 cycles at peak-negative pressures of 0.6 to 6.3 MPa. 1000 valid single-bubble backscatter events at each exposure condition were digitized and then analyzed for 20-MHz subharmonic content. The data showed that the subharmonic response was initiated between 5 and 10 cycles and the likelihood of a subharmonic event increased as the number of cycles increased. A subharmonic backscatter response was most likely at 3.9 MPa for the 3.4 mum agent and 1.7 MPa for the 0.56 and 1.1 mum agents. The increased pressure for subharmonic activity for larger agent was consistent with its larger size.     

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。     

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.     

Immunospecific NMR contrast agents

Most NMR contrast agents suggested to date have been paramagnetic. These agents, which include the transition and lanthanide metal ions as well as stable organic free radicals, do not provide effective contrast at concentrations much below 1 mM. However, the use of macromolecular ferromagnetic and superparamagnetic particles provides, for the first time, an NMR relaxation agent that is effective at subnanomolar concentrations. Two different sized superparamagnetic particles have been coupled to monoclonal antibodies with high affinity for a neuroblastoma-specific cell surface antigen. The specific binding of these particles, both in vivo and in vitro is demonstrated and the consequences for immunospecific NMR contrast are discussed.     

Subharmonic imaging of contrast agents

Ultrasound contrast agents promise to improve the sensitivity and specificity of diagnostic ultrasound imaging. It is of great importance to adapt ultrasound equipment for optimal use with contrast agents e.g., by exploiting the nonlinear properties of the contrast microbubbles. Harmonic imaging is one technique that has been extensively studied and is commercially available. However, harmonic imaging is associated with problems, due to second harmonic generation and accumulation within the tissue itself. Given the lack of subharmonic generation in tissue, one alternative is the creation of subharmonic images by transmitting at the fundamental frequency (fo) and receiving at the subharmonic (fo/2). Subharmonic imaging should have a much better lateral resolution and may be suitable for scanning deep-lying structures owing to the higher transmit frequency and the much smaller attenuation of scattered subharmonic signals. In this paper, we will review different aspects of subharmonic imaging including implementation, in-vitro gray-scale imaging and subharmonic aided pressure estimation.     

Gadolinium-labeled liposomes containing amphiphilic Gd-DTPA derivatives of varying chain length: targeted MRI contrast enhancement agents for the liver

Paramagnetic liposomal contrast agents were synthesized and utilized for selective augmentation of T1 MR imaging of the livers of normal Balb/c mice. Amphiphilic gadolinium complexes, which mimic phospholipids, were incorporated into the lamella of small unilamellar liposomes (SUV) such that they become an integral part of its surface. The amphiphilic complexing agents consisted of DTPA reagents in which a pair of alkyl chains of varying lengths are attached via amide linkages. The in vivo lifetimes of the amphiphilic agents were found to be dependent on the chain length of the alkyl groups.     

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.     

Ambient pressure dependence of the ultra-harmonic response from contrast microbubbles

Sub-harmonic response from ultrasound contrast agent microbubbles has been demonstrated to be an effective modality for noninvasive pressure measurement. In the present study, the dependence of ultra-harmonic response on the ambient overpressure was investigated by both experimental measurements and simulations. In the measurements, the microbubbles were exposed to Gaussian pulses with varied driving frequencies and pulse lengths, at an acoustic pressure of 0.3 MPa. The amplitudes of sub- and ultra-harmonic components were measured when the ambient overpressures varied from 0-25 kPa. At the driving frequency of 1.33 MHz, the ultra-harmonic energy decreased but the sub-harmonic energy increased with the increasing overpressure; while at the driving frequency of 4 MHz, both the sub- and ultra-harmonic components showed the same tendency that the corresponding energy decreased as the overpressure was increased. A 4-MHz Gaussian pulse with 64 cycles could provide an ultra-harmonic response with both good ambient pressure sensitivity and high linearity. Furthermore, the effects of shell parameters of a microbubble on the generation of ultra- and sub-harmonic responses were discussed based on simulations using Marmottant's model. This study suggests that the ultra-harmonic response from contrast microbubbles might be applicable for noninvasive pressure measurement.     

Enhanced ferrite nanoparticles as MRI contrast agents

Enhanced ferrite nanoparticles are a new class of contrast agents for magnetic resonance imaging (MRI). The enhanced ferrites are synthesized by reverse micelles technique to form iron core and oxide or ferrite shell preventing further oxidation of the nanoparticles. The nanoparticles are further functionalized using dopamine and PEG-600 to increase the solubility of the high magnetic moment nanoparticles. ¹H relaxation measurements of aqueous solutions of the nanoparticles were conducted at 2.4 T. The relaxivities r₁ and r₂, representing the slopes of these curves, are 7.19 and 9.96 s⁻¹ mM⁻¹, respectively. These values should be compared with relaxivities of 4–5 s⁻¹ mM⁻¹ corresponding to commonly used commercial contrast agents in human MR examinations.     

Potential contrast agents for magnetic resonance imaging

The development ofmagneto-pharmaceuticals plays an important role in the extension of nuclear magnetic resonance into diagnostic medicine. That is the reason why fundamental investigations leading to new insights into NMR contrast agents are presently being considered. The synthesis and the proton relaxation rates of some new contrast agents are presented. The high values ofR ₁, andR ₂ relaxivities of the compounds studied by us are promising for various and novel applications.