The impact of bubbles on measurement of drug release from echogenic liposomes

Echogenic liposomes (ELIP) encapsulate gas bubbles and drugs within lipid vesicles, but the mechanisms of ultrasound-mediated drug release from ELIP are not well understood. The effect of cavitation activity on drug release from ELIP was investigated in flowing solutions using two fluorescent molecules: a lipophilic drug (rosiglitazone) and a hydrophilic drug substitute (calcein). ELIP samples were exposed to pulsed Doppler ultrasound from a clinical diagnostic ultrasound scanner at pressures above and below the inertial and stable cavitation thresholds. Control samples were exposed to a surfactant, Triton X-100 (positive control), or to flow alone (negative control). Fluorescence techniques were used to detect release. Encapsulated microbubbles reduced the measured fluorescence intensity and this effect should be considered when assessing drug release from ELIP. The origin of this effect is not specific to ELIP. Release of rosiglitazone or calcein compared to the negative control was only observed with detergent treatment, but not with ultrasound exposure, despite the presence of stable and inertial cavitation activity. Release of rosiglitazone or calcein from ELIP exposed to diagnostic ultrasound was not observed, even in the presence of cavitation activity. Ultrasound-mediated drug delivery strategies with ELIP will thus rely on passage of the drug-loaded liposomes to target tissues.     

Barrier-breaking effects of ultrasonic cavitation for drug delivery and biomarker release

Recently, emerging evidence has demonstrated that cavitation actually creates important bidirectional channels on biological barriers for both intratumoral drug delivery and extratumoral biomarker release. To promote the barrier-breaking effects of cavitation for both therapy and diagnosis, we first reviewed recent technical advances of ultrasound and its contrast agents (microbubbles, nanodroplets, and gas-stabilizing nanoparticles) and then reported the newly-revealed cavitation physical details. In particular, we summarized five types of cellular responses of cavitation in breaking the plasma membrane (membrane retraction, sonoporation, endocytosis/exocytosis, blebbing and apoptosis) and compared the vascular cavitation effects of three different types of ultrasound contrast agents in breaking the blood-tumor barrier and tumor microenvironment. Moreover, we highlighted the current achievements of the barrier-breaking effects of cavitation in mediating drug delivery and biomarker release. We emphasized that the precise induction of a specific cavitation effect for barrier-breaking was still challenged by the complex combination of multiple acoustic and non-acoustic cavitation parameters. Therefore, we provided the cutting-edge in-situ cavitation imaging and feedback control methods and suggested the development of an international cavitation quantification standard for the clinical guidance of cavitation-mediated barrier-breaking effects.     

Effect of local dual frequency sonication on drug distribution from polymeric nanomicelles

To overcome the side effects caused by systemic administration of doxorubicin, nanosized polymeric micelles were used in combination with dual frequency ultrasonic irradiation. These micelles release the drug due to acoustic cavitation, which is enhanced in dual frequency ultrasonic fields. To form the drug-loaded micelles, Pluronic P-105 copolymer was used, and doxorubicin was physically loaded into stabilized micelles with an average size of 14 nm. In this study, adult female Balb/C mice were transplanted with spontaneous breast adenocarcinoma tumors and were injected with a dose of 1.3 mg/kg doxorubicin in one of three forms: free doxorubicin, micellar doxorubicin without sonication and micellar doxorubicin with sonication. To increase cavitation yield, the tumor region was sonicated for 2.5 min at simultaneous frequencies of 3 MHz (I(SATA)=2 W/cm(2)) and 28 kHz (I(SATA)=0.04 W/cm(2)). The animals were sacrificed 24h after injection, and their tumor, heart, spleen, liver, kidneys and plasma were separated and homogenized. The drug content in the tissues was determined using tissue fluorimetry (350 nm excitation and 560 nm emission), and standard drug dose curves were obtained for each tissue. The results show that in the group that received micellar doxorubicin with sonication, the drug concentration in the tumor tissue was significantly higher than in the free doxorubicin injection group (8.69 times) and the micellar doxorubicin without sonication group (2.60 times). The drug concentration in other tissues was significantly lower in the micellar doxorubicin with sonication group relative to the free doxorubicin (3.35 times) and the micellar drug without sonication (2.48 times) groups (p<0.05). We conclude that dual frequency sonication improves drug release from micelles and increases the drug uptake by tumors due to sonoporation. The proposed drug delivery system creates an improved treatment capability while reducing systemic side effects caused by drug uptake in other tissues.     

Single-transducer dual-frequency ultrasound generation to enhance acoustic cavitation

Dual- or multiple-frequency ultrasound stimulation is capable of effectively enhancing the acoustic cavitation effect over single-frequency ultrasound. Potential application of this sonoreactor design has been widely proposed such as on sonoluminescence, sonochemistry enhancement, and transdermal drug release enhancement. All currently available sonoreactor designs employed multiple piezoelectric transducers for generating single-frequency ultrasonic waves separately and then these waves were mixed and interfered in solutions. The purpose of this research is to propose a novel design of generating dual-frequency ultrasonic waves with single piezoelectric elements, thereby enhancing acoustic cavitation. Macroscopic bubbles were detected optically, and they were quantified at either a single-frequency or for different frequency combinations for determining their efficiency for enhancing acoustic cavitation. Visible bubbles were optically detected and hydrogen peroxide was measured to quantify acoustic cavitation. Test water samples with different gas concentrations and different power levels were used to determine the efficacy of enhancing acoustic cavitation of this design. The spectrum obtained from the backscattered signals was also recorded and examined to confirm the occurrence of stable cavitation. The results confirmed that single-element dual-frequency ultrasound stimulation can enhance acoustic cavitation. Under certain testing conditions, the generation of bubbles can be enhanced up to a level of five times higher than the generation of bubbles in single-frequency stimulation, and can increase the hydrogen peroxide production up to an increase of one fold. This design may serve as a useful alternative for future sonoreactor design owing to its simplicity to produce dual- or multiple-frequency ultrasound.     

Stability of alteplase in presence of cavitation

Several experimental studies have demonstrated that ultrasound (US) can accelerate enzymatic fibrinolysis and this effect is further enhanced in the presence of ultrasound contrast agents (UCA). Although UCA have been shown to be safe when administered to ischemic stroke patients, safety information of these agents in the thrombolysis setting is limited. Therefore, in this study we investigated potential adverse effects of acoustic cavitation generated by UCA on alteplase (t-PA), the drug used for treatment of ischemic stroke patients. A volume of 0.9 mL of alteplase was dispensed into a custom-made polyester sample tube. For treatments in the presence or absence of cavitation either 0.1 mL Optison or phosphate buffer saline was combined with alteplase. Three independent samples of each treatment group were exposed to ultrasound of 2 MHz frequency at three different peak negative acoustic pressures of 0.5, 1.7, and 3.5 MPa for a duration of 60 min. All treatments were carried out in a cavitation detection system which was used to insonify the samples and record acoustic emissions generated within the sample. After ultrasound exposure, the treated samples and three untreated drug samples were tested for their enzymatic activity using a chromogenic substrate. The insonified samples containing Optison demonstrated cavitational activity proportional to acoustic pressure. No significant cavitation activity was observed in the absence of Optison. Enzymatic activity of alteplase in both insonified groups was comparable to that in the control group. These tests demonstrated that exposure of alteplase to 60 min of 2 MHz ultrasound at acoustic pressures ranging from 0.5 MPa to 3.5 MPa, in the presence or absence of Optison had no adverse effects on the stability of this therapeutic compound.     

A transfer efficiency model for ultrasound mediated drug/gene transferring into cells

Ultrasound is a very promising technology to mediated drug/gene transferring into cells. However the relations between cell experimental conditions and results have been still unknown. It seriously impeded the development of the technology. In the article, a transfer efficiency model for ultrasound mediated drug/gene transferring into cells in stable cavitation was constructed. To testify the model, the numerical results were compared with the cell experimental data from six literatures in the entirely different experimental conditions. The numerical results fit the cell experimental data well. Despite simplifications and limitations, the model for the first time established the relationship between the cell experimental results about transfer efficiency and the conditions including ultrasound, microbubble and cells in stable cavitation.     

Leveraging re-chargeable nanobubbles on amine-functionalized ZnO nanocrystals for sustained ultrasound cavitation towards echographic imaging

Nanoparticles able to promote inertial cavitation when exposed to focused ultrasound have recently gained much attention due to their vast range of possible applications in the biomedical field, such as enhancing drug penetration in tumor or supporting ultrasound contrast imaging. Due to their nanometric size, these contrast agents could penetrate through the endothelial cells of the vasculature to target tissues, thus enabling higher imaging resolutions than commercial gas-filled microbubbles. Herein, Zinc Oxide NanoCrystals (ZnO NCs), opportunely functionalized with amino-propyl groups, are developed as novel nanoscale contrast agents that are able, for the first time, to induce a repeatedly and over-time sustained inertial cavitation as well as ultrasound contrast imaging. The mechanism behind this phenomenon is investigated, revealing that re-adsorption of air gas nanobubbles on the nanocrystal surface is the key factor for this re-chargeable cavitation. Moreover, inertial cavitation and significant echographic signals are obtained at physiologically relevant ultrasound conditions (MI < 1.9), showing great potential for low side-effects in in-vivo applications of the novel nanoscale agent from diagnostic imaging to gas-generating theranostic nanoplatforms and to drug delivery.     

Correlation between microbubble-induced acoustic cavitation and hemolysis in vitro

Microbubbles promise to enhance the efficiency of ultrasound-mediated drug delivery and gene therapy by taking advantage of artificial cavitation nuclei.The purpose of this study is to examine the ultrasound-induced hemolysis in the application of drug delivery in the presence of microbubbles.To achieve this goal,human red blood cells mixed with microbubbles were exposed to 1-MHz pulsed ultrasound.The hemolysis level was measured by a flow cytometry,and the cavitation dose was detected by a passive cavitation detecting system.The results demonstrate that larger cavitation dose would be generated with the increase of acoustic pressure,which might give rise to the enhancement of hemolysis.Besides the experimental observations,the acoustic pressure dependence of the radial oscillation of microbubble was theoretically estimated.The comparison between the experimental and calculation results indicates that the hemolysis should be highly correlated to the acoustic cavitation.     

Cavitation dose in an ultrasonic cleaner and its dependence on experimental parameters

The aluminum foil erosion method is widely used in cavitation activity studies of ultrasonic cleaners. However, owing to its limited sensitivity, it is difficult to observe the effects of various experimental parameters on the cavitation activity using this method. In the present work, a higher-sensitivity method for quantifying cavitation activity as a cavitation dose based on passive cavitation detection was presented. The influences of various factors (e.g., insonation duration, driving power, gas content, temperature and cleaning agents) were studied for this system. The results showed that the cavitation dose became unstable over long insonation times, and that the instability was more significant at high power. Generally, the cavitation activity could be enhanced by increasing the power, gas content, and the concentration of a cleaning agent. However, due to the exhaustion of the cavitation gas nuclei, the cavitation activity might tune to saturate of even decrease slightly when some impact parameters (e.g., acoustic driving power, gas content and the concentration of the cleaning agent) are above a certain level of each of these parameters.     

Quantifying OH radical generation in hydrodynamic cavitation via coumarin dosimetry: Influence of operating parameters and cavitation devices

Hydrodynamic cavitation (HC) has been extensively investigated for effluent treatment applications. Performance of HC devices or processes is often reported in terms of degradation of organic pollutants rather than quantification of hydroxyl (OH) radicals. In this study, generation of OH radicals in vortex based cavitation device using coumarin dosimetry was quantified. Coumarin was used as the chemical probe with an initial concentration of 100 µM (15 ppm). Generation of OH radicals was quantified by analysing generated single hydroxylated products. The influence of operating parameters such as pH and type of acid used to adjust pH, dissolved oxygen, and inlet and outlet pressures was investigated. Acidic pH was found to be more conducive for generating OH radicals and therefore subsequent experiments were performed at pH of 3. Sulphuric acid was found to be more than three times effective than hydrochloric acid in generating OH radicals. Effect of initial levels of dissolved oxygen was found to influence OH radical generation. Performance of vortex based cavitation device was then compared with other commonly used cavitation devices based on orifice and venturi. The vortex based cavitation device was found to outperform the orifice and venturi based devices in terms of initial per-pass factor. Influence of device scale (nominal flow rate through the device) on performance was then evaluated. The results presented for these devices unambiguously quantifies their cavitational performance. The presented results will be useful for evaluating computational models and stimulate further development of predictive computational models in this challenging area.     

Towards an understanding and control of cavitation activity in 1 MHz ultrasound fields

Various industrial processes such as sonochemical processing and ultrasonic cleaning strongly rely on the phenomenon of acoustic cavitation. As the occurrence of acoustic cavitation is incorporating a multitude of interdependent effects, the amount of cavitation activity in a vessel is strongly depending on the ultrasonic process conditions. It is therefore crucial to quantify cavitation activity as a function of the process parameters. At 1 MHz, the active cavitation bubbles are so small that it is becoming difficult to observe them in a direct way. Hence, another metrology based on secondary effects of acoustic cavitation is more suitable to study cavitation activity. In this paper we present a detailed analysis of acoustic cavitation phenomena at 1 MHz ultrasound by means of time-resolved measurements of sonoluminescence, cavitation noise, and synchronized high-speed stroboscopic Schlieren imaging. It is shown that a correlation exists between sonoluminescence, and the ultraharmonic and broadband signals extracted from the cavitation noise spectra. The signals can be utilized to characterize different regimes of cavitation activity at different acoustic power densities. When cavitation activity sets on, the aforementioned signals correlate to fluctuations in the Schlieren contrast as well as the number of nucleated bubbles extracted from the Schlieren Images. This additionally proves that signals extracted from cavitation noise spectra truly represent a measure for cavitation activity. The cyclic behavior of cavitation activity is investigated and related to the evolution of the bubble populations in the ultrasonic tank. It is shown that cavitation activity is strongly linked to the occurrence of fast-moving bubbles. The origin of this “bubble streamers” is investigated and their role in the initialization and propagation of cavitation activity throughout the sonicated liquid is discussed. Finally, it is shown that bubble activity can be stabilized and enhanced by the use of pulsed ultrasound by conserving and recycling active bubbles between subsequent pulsing cycles.     

超声空化现象影响因素的实验研究

超声空化在许多不同的学科和工业生产中有着广泛的应用。超声空化的应用与声场的分布及空化的机理密切相关,精准地反映空化场和空化机理是超声空化技术实际应用的关键。该文通过分析采集的声信号和金属箔膜空蚀法对空化区域随液位发生变化的现象进行研究,并利用Matlab对金属箔膜空蚀程度量化。实验发现,超声波会在液面与实验箱体底部形成驻波场。在某一液体温度下,随着液位高度的变化,超声空化现象的出现具有周期性。并且,在同一液位下,当超声功率改变时,空化区域强度分布情况随之改变。小功率时各空化区域空化强度分布均匀,当功率增大到一定时,会出现空化屏蔽现象。该研究为超声清洗设备的改良提供了借鉴,对进一步认识和利用超声空化效应具有重要意义。     

Impact of cavitation on lesion formation induced by high intensity focused ultrasound

High intensity focused ultrasound(HIFU) has shown a great promise in noninvasive cancer therapy. The impact of acoustic cavitation on the lesion formation induced by HIFU is investigated both experimentally and theoretically in transparent protein-containing gel and ex vivo liver tissue samples. A numerical model that accounts for nonlinear acoustic propagation and heat transfer is used to simulate the lesion formation induced by the thermal effect. The results showed that lesions could be induced in the samples exposed to HIFU with various acoustic pressures and pulse lengths. The measured areas of lesions formed in the lateral direction were comparable to the simulated results, while much larger discrepancy was observed between the experimental and simulated data for the areas of longitudinal lesion cross-section. Meanwhile,a series of stripe-wiped-off B-mode pictures were obtained by using a special imaging processing method so that HIFUinduced cavitation bubble activities could be monitored in real-time and quantitatively analyzed as the functions of acoustic pressure and pulse length. The results indicated that, unlike the lateral area of HIFU-induced lesion that was less affected by the cavitation activity, the longitudinal cross-section of HIFU-induced lesion was significantly influenced by the generation of cavitation bubbles through the temperature elevation resulting from HIFU exposures. Therefore, considering the clinical safety in HIFU treatments, more attention should be paid on the lesion formation in the longitudinal direction to avoid uncontrollable variation resulting from HIFU-induced cavitation activity.     

The Relationship of Cavitation to the Negative Acoustic Pressure Amplitude in Ultrasonic Therapy

The relationship between the cavitation and acoustic peak negative pressure in the high-intensity focused ultrasound(HIFU)Held is analyzed in water and tissue phantom.The peak negative pressure at the focus is determined by a hybrid approach combining the measurement with the simulation.The spheroidal beam equation is utilized to describe the nonlinear acoustic propagation.The waveform at the focus is measured by a fiber optic probe hydrophone in water.The relationship between the source pressure amplitude and the excitation voltage is determined by fitting the measured ratio of the second harmonic to the fundamental component at the focus,based on the model simulation.Then the focal negative pressure is calculated for arbitrary voltage excitation in water and tissue phantom.A portable B-mode ultrasound scanner is applied to monitor HIFU-induced cavitation in real time,and a passive cavitation detection(PCD)system is used to acquire the bubble scattering signals in the HIFU focal volume for the cavitation quantification.The results show that:(1)unstable cavitation starts to appear in degassed water when the peak negative pressure of HIFU signals reaches 13.5 MPa;and(2)the cavitation activity can be detected in tissue phantom by B-mode images and in the PCD system with HIFU peak negative pressures of 9.0 MPa and 7.8 MPa,respectively,which suggests that real-time B-mode images could be used to monitor the cavitation activity in two dimensions,while PCD systems are more sensitive to detect scattering and emission signals from cavitation bubbles.     

超声空化引起界面湍动促进的传质机理

本文分析了超声空化引起界面湍动对传质过程的影响,提出了相界面上超声空化气泡析出增强边界层液体湍动并促进传质的机理,在传质理论和流体动力学原理的基础上,建立了超声空化引起界面湍动促进的传质机理模型,获得了超声空化引起界面湍动促进的传质系数表达式。实验结果验证了模型的合理性。该模型既证实了超声对传质有强化效应,又对传质过程有很好的预测功能,为工业化提供了理论依据。     

伴随水力空化现象的对流换热数值研究

以水为介质,建立了液体流动的混合物多相流模型及空化模型,运用CFD方法对水平圆管内伴随有水力空化现象的受迫对流换热过程进行了数值研究,详细分析了管道入口压力、入口温度和限流孔与管道直径比等因素对水力空化及对流换热过程的影响规律。数值模拟结果表明,空化现象出现在圆管喉部(限流孔)壁面附近区域;与相同流量下无空化时的传热相比,在发生空化现象的区域,传热壁面被蒸汽所覆盖导致传热急剧恶化,而在远离空化发生区域的下游位置,由于空化的扰流作用使得加热壁面与流体之间的传热得到明显改善。     

Acoustic characterization of cavitation intensity: A review

Cavitation intensity is used to describe the activity of cavitation, and several methods are developed to identify the intensity of cavitation. This work aimed to provide an overview and discussion of the several existing characterization methods for cavitation intensity, three acoustic approaches for charactering cavitation were discussed in detail. It was showed that cavitation noise spectrum is too complex and there are some differences and disputes on the characterization of cavitation intensity by cavitation noise. In this review, we recommended a total cavitation noise intensity estimated via the integration of real cavitation noise spectrum over full frequency domain instead of artificially adding inaccurate filtering processing.     

Cavitation technology – A greener processing technique for the generation of pharmaceutical nanoemulsions

Novel nanoemulsion-based drug delivery systems (DDS) have been proposed as alternative and effective approach for the delivery of various types of poorly water-soluble drugs in the last decade. This nanoformulation strategy significantly improves the cell uptake and bioavailability of numerous hydrophobic drugs by increasing their solubility and dissolution rate, maintaining drug concentration within the therapeutic range by controlling the drug release rate, and reducing systemic side effects by targeting to specific disease site, thus offering a better patient compliance. To date, cavitation technology has emerged to be an energy-efficient and promising technique to generate such nanoscale emulsions encapsulating a variety of highly potent pharmaceutical agents that are water-insoluble. The micro-turbulent implosions of cavitation bubbles tear-off primary giant oily emulsion droplets to nano-scale, spontaneously leading to the formation of highly uniform drug contained nanodroplets. A substantial body of recent literatures in the field of nanoemulsions suggests that cavitation is a facile, cost-reducing yet safer generation tool, remarkably highlighting its industrial commercial viability in the development of designing novel nanocarriers or enhancing the properties of existing pharmaceutical products. In this review, the fundamentals of nanoemulsion and the principles involved in their formation are presented. The underlying mechanisms in the generation of pharmaceutical nanoemulsion under acoustic field as well as the advantages of using cavitation compared to the conventional techniques are also highlighted. This review focuses on recent nanoemulsion-based DDS development and how cavitation through ultrasound and hydrodynamic means is useful to generate the pharmaceutical grade nanoemulsions including the complex double or submicron multiple emulsions.     

叶片背面不连续凸起对轴流泵空化的控制研究

为控制轴流泵空化的发生与发展,提出了一种在轴流泵叶片背面布置不连续凸起结构的方案。基于ANSYS CFX软件,对350ZQ-125-30型潜水轴流泵进行数值模拟,对比分析改进模型与原模型流道内压力、湍动能、空泡及流线分布的变化.结果表明,改进后叶片背面低压区域减小,工作面高压区域明显增大,临界空化余量降低;在空化的各个阶段空泡体积分数都有所减小,其中空化初生阶段最为明显,相比原模型减小了38.3%,有效地控制了空化的发生,空化性能明显提升;在空化发展阶段,凸起的存在能够有效阻挡回射流的冲击,控制了空泡的脱落,同时抑制了叶轮流道内梢涡空化的产生。