Microbubble Suspension Research Articles

An acoustic rectifier

The detection of acoustic signals is of relevance for a range of practical applications, for example in medical diagnostics. However, whereas rectification of electric current and other energy forms such as thermal flux has been demonstrated, acoustic rectification has not yet been achieved. Here, on the basis of the earlier theoretical proposal of an 'acoustic diode', we present the first experimental demonstration of a rectified energy flux of acoustic waves. A one-dimensional acoustic rectifier is fabricated by coupling a superlattice with a layer of ultrasound contrast agent microbubble suspension. A significant rectifying effect is observed within two frequency bands at locations that agree well with theoretical predictions. Following optimization of the concentration of the microbubble suspension, rectifying ratios can be as high as ~10(4). This realization of an acoustic rectifier should have substantial practical significance, for example in the focusing of ultrasound in medical applications.

Effect of Microbubble Size on Fundamental Mode High Frequency Ultrasound Imaging in Mice

High-frequency ultrasound imaging using microbubble (MB) contrast agents is becoming increasingly popular in pre-clinical and small animal studies of anatomy, flow and vascular expression of molecular epitopes. Currently, in vivo imaging studies rely on highly polydisperse microbubble suspensions, which may provide a complex and varied acoustic response. To study the effect of individual microbubble size populations, microbubbles of 1–2 μm, 4–5 μm and 6–8 μm diameter were isolated using the technique of differential centrifugation. Size-selected microbubbles were imaged in the mouse kidney over a range of concentrations using a Visualsonics Vevo 770 ultrasound imaging system (Visualsonics, Toronto, Ontario, Canada) with a 40-MHz probe in fundamental mode. Results demonstrate that contrast enhancement and circulation persistence are strongly dependent on microbubble size and concentration. Large microbubbles (4–5 and 6–8 μm) strongly enhanced the ultrasound image with positive contrast, while 1–2 μm microbubbles showed little enhancement. For example, the total integrated contrast enhancement, measured by the area under the time-intensity curve (AUC), increased 16-fold for 6–8 μm diameter microbubbles at 5 × 10 7 MB/bolus compared with 4–5 μm microbubbles at the same concentration. Interestingly, 1–2 μm diameter microbubbles, at any concentration, did not measurably enhance the integrated ultrasound signal at tissue depth, but did noticeably attenuate the signal, indicating that they had a low scattering-to-attenuation ratio. When concentration matched, larger microbubbles were more persistent in circulation. However, when volume matched, all microbubble sizes had a similar circulation half-life. These results indicated that dissolution of the gas core plays a larger role in contrast elimination than filtering by the lungs and spleen. The results of this study show that microbubbles can be tailored for optimal contrast enhancement in fundamental mode imaging. (E-mail: mb2910@columbia.edu)

J0203-2-6 集束超音波によるソナゾイドの破壊実験([J0203-2]ドラッグデリバリーシステム:技術開発と医療応用(2))

For destroying Sonazoid microbubbles 1 MHz - ultrasound waves was irradiated by employing a concave ultrasound probe with the vibrating diameter of 40 mm and the arithmetic focal length of 80 mm. The probe was set in distilled water at temperature of 25 ℃. A suspension of Sonazoid ultrasound contrast agent diluted with distilled water with the volumetric concentration of 5 % [v/v] was shut in a cuvette which was positioned at x_<c,b> measured from the bottom of the ultrasound probe. It was found that Sonazoid microbubbles were rapidly destroyed and the survival number was reduced to 40 % of the original number at the exposure time of 100 ms. When a cuvette containing suspension of Sonazoid microbubbles was set at the arithmetic focal point, a limited volume of Sonazoid suspension was exposed to the ultrasound waves, resulting in a relatively larger survival number of Sonazoid microbubbles. Sonazoid destruction was observed by taking snapshots using a nano-pulse light with the pulse duration of 180 ns as a light source.

Novel preparation techniques for controlling microbubble uniformity: a comparison

In recent years, there has been increasing interest in the use of coated microbubbles as vehicles for ultrasound mediated targeted drug delivery. This application requires a high degree of control over the size and uniformity of microbubbles, in order to ensure accurate dosing of a given drug and to maximise delivery efficiency. Similarly, as more advanced imaging techniques are developed which exploit the complex nonlinear features of the microbubble signal and/or enable quantification of tissue perfusion, the ability to predetermine the acoustic response of a microbubble suspension is becoming increasingly important. Consequently, a number of new preparation technologies have been developed to meet the demand for improved control over microbubble characteristics. The aim of the work described in this paper was to compare a conventional microbubble preparation technique, sonication, with two more recent methods: coaxial electrohydrodynamic atomisation and microfluidic (T-junction) processing, in terms of their ability to produce bubbles which are sufficiently small and stable for in vivo use, microbubble uniformity, relative production rates and other practical and economic considerations.

Enhancement of aerobic biodegradation in an oxygen-limiting environment using a saponin-based microbubble suspension

This study investigated the ability of a saponin-based microbubble suspension to enhance aerobic biodegradation of phenanthrene by subsurface delivery. As the microbubble suspension flowed through a sand column pressure buildup and release was repeatedly observed, which delivered oxygen to the less permeable regions. Burkholderia cepacia RPH1, a phenanthrene-degrading bacterium, was mainly transported in a suspended form in the microbubble suspension. When three pore volumes of the microbubble suspension containing B. cepacia RPH1 was introduced into a column contaminated with phenanthrene (100 mg/kg), the oxygen content declined to 5% from an initial value of 20% within 5 days and correspondingly, 34.4% of initial phenanthrene was removed in 8 days. The addition of two further three pore volumes enhanced the biodegradation efficiency by a factor of 2.2. Our data suggest that a saponin-based microbubble suspension could be a potential carrier for enhancing the aerobic biodegradation under an oxygen-limiting environment.

Microbubbles as a novel contrast agent for brain MRI

Gas-filled microbubbles have the potential to become a unique MR contrast agent due to their magnetic susceptibility effect, biocompatibility and localized manipulation via ultrasound cavitation. In this study, two types of microbubbles, custom-made albumin-coated microbubbles (A-MB) and a commercially available lipid-based clinical ultrasound contrast agent (SonoVue®), were investigated with in vivo dynamic brain MRI in Sprague–Dawley rats at 7 T. Microbubble suspensions (A-MB: 0.2 mL of ∼4% volume fraction; SonoVue®: 0.2 mL of ∼3.5% volume fraction) were injected intravenously. Transverse relaxation rate enhancements (ΔR2⁎) of 2.49±1.00 s−1 for A-MB and 2.41±1.18 s−1 for SonoVue® were observed in the brain (N=5). Brain ΔR2⁎ maps were computed, yielding results similar to the cerebral blood volume maps obtained with a common MR blood pool contrast agent. Microbubble suspension ΔR2⁎ was measured for different volume fractions. These results indicate that gas-filled microbubbles can serve as an intravascular contrast agent for brain MRI at high field. Such capability has the potential to lead to real-time MRI guidance in various microbubble-based drug delivery and therapeutic applications in the central nervous system.

Fabrication of indocyanine green encapsulated biodegradable microbubbles for structural and functional imaging of cancer

We developed a novel dual-modal contrast agent for the structural and functional imaging of cancer. The contrast agent was fabricated by encapsulating indocyanine green (ICG) in poly(lactic-co-glycolic acid) (PLGA) microbubbles using a modified double-emulsion method. More stabilized absorption and fluorescence emission characteristics were observed for aqueous and plasma suspensions of ICG-encapsulated microbubbles. The technical feasibility of concurrent structural and functional imaging was demonstrated through a series of benchtop tests in which the aqueous suspension of ICG-encapsulated microbubbles was injected into a transparent tube embedded in an Intralipid phantom at different flow rates and concentrations. Concurrent fluorescence imaging and B-mode ultrasound imaging successfully captured the changes of microbubble flow rate and concentration with high linearity and accuracy. One potential application of the proposed ICG-encapsulated PLGA microbubbles is for the identification and characterization of peritumoral neovasculature for enhanced coregistration between tumor structural and functional boundaries in ultrasound-guided near-infrared diffuse optical tomography.

Automatic liquid injection system and method

A power assisted method and injector device for controllably delivering to patients a dispersion medicament or diagnostically active agent, the homogeneity of which is preserved throughout delivery. Diagnostically active agents disclosed are gas microbubble suspensions useful in ultrasonic diagnostic imaging and liposomal formulations in which liposome vesicles are loaded with iodinated compounds.

Microbubble Destruction During Intravenous Administration: A Preliminary Study

The concentration and size distribution of microbubble suspensions are important parameters for both diagnostic and therapeutic applications. The aim of this preliminary study was to investigate the relationship between changes in the microbubble population and the administration process variables, specifically syringe inner diameter, needle inner diameter, volume flow rate and the liquid in which the microbubbles are suspended. It was found that reducing either the syringe or needle inner diameter produced large reductions in microbubble concentration during administration, as much as 99.9% for needle inner diameters <0.24 mm. Increasing the volume flow rate up to 3 mL/min and changing the suspending fluid from distilled water to glycerol, however, were both found to reduce the degree of microbubble destruction. Further work is needed to fully explain these observations. However, investigation of the response of microbubbles to changes in hydrostatic pressure only, indicated that this was unlikely to be the main mechanism of destruction and hence that shear stress was a more important factor. Comparison with findings from another recent study of microbubble stability indicated that microbubble size, concentration and composition were also important parameters and should be taken into account in designing administration procedures for microbubble agents. It was concluded that current procedures should be reviewed, particularly for therapeutic applications, and that the results should also be taken into account when assessing the accuracy of microbubble size distribution measurements obtained using automatic particle sizing equipment in which microbubbles are made to flow under pressure. (E-mail: e_stride@meng.ucl.ac.uk)

Pressure-Dependent Attenuation and Scattering of Phospholipid-Coated Microbubbles at Low Acoustic Pressures

Previous optical studies have shown threshold behavior of single-contrast agent microbubbles. Below the acoustic pressure threshold, phospholipid-coated microbubbles with sizes <5.0 μm in diameter oscillate significantly less than above the threshold pressure. Previous studies also revealed an acoustic pressure-dependent attenuation of ultrasound by microbubble contrast agents. In this study, we investigated whether pressure-dependent acoustic behavior may be explained by threshold behavior. For this purpose, pressure-dependent attenuation and scattering of a phospholipid-coated contrast agent were measured. Transmit frequencies between 1.5 and 6.0 MHz and acoustic pressures between 5 and 200 kPa were applied. Unlike the galactose-based contrast agent Levovist, the phospholipid-coated contrast agent BR14 showed a pressure-dependent attenuation. In addition, it was found that filtered suspensions with only microbubbles <3.0 μm in diameter show more pressure-dependent attenuation behavior than native suspensions of phospholipid-coated microbubbles. For the scattering measurements conducted at 3.0 MHz, the native suspension did not show any pressure-dependent behavior. However, the filtered suspension responded highly nonlinearly. Between 30 and 150 kPa, 16 dB additional scattered power was obtained. We concluded that threshold behavior of phospholipid-coated microbubbles results in pressure-dependent attenuation and scattering. (E-mail: m.emmer@erasmusmc.nl)

Stability and rheological behavior of concentrated monodisperse food emulsifier coated microbubble suspensions

Nearly monodispersed populations of microbubbles were produced using flow focusing with a food grade emulsifier. The microbubbles produced by this technique have diameters in the range of 120–200 μm. The flow focusing device uses metered streams of air and liquid to produce a jet that periodically pinches to make individual microbubbles. The size of the microbubbles can be controlled by changing the relative flow rates of the gas and the liquid. The emulsifier consists of a mixture of monoglycerides, diglycerides and sodium stearoyl lactylate in combination with polyethylene glycol (PEG)-40 stearate. The emulsifier forms a thin shell that stabilizes the microbubbles. The microbubbles are stable over time with their sizes remaining roughly constant over 2 h. Such stability allows suspensions of microbubbles to be formed and their rheological properties tested. The sizes of the microbubbles are also monitored off-line while testing, examining the effect of shearing on the bubble sizes, as well as their stability over time. These results show that the microbubble suspensions are viscoelastic and exhibit power law behavior. The relationship between the air fraction of the suspension and fluid viscosity is determined.

Generation of a Monodisperse Microbubble Jet in Microgravity

A new method to create a jet of a virtually monodisperse microbubble suspension of prescribed bubble size into a quiescent cavity is proposed. The method is insensitive to gravity and is based on the creation of a slug flow at a T junction in a capillary tube before injection. We develop a theoretical analysis that establishes the validity and efficiencyofthe method,ascontrolledbythecrossflowWebernumber,andyields asimple explicitprediction forthe bubble size in terms of the injection parameters. The method operates efficiently for small Weber numbers, yet it generates small bubbles of very uniform size. The reduced size dispersion is also explained within the theoretical model. The method of bubble formation, injection, and spreading by the resulting turbulent jet is validated experimentally in 4.7 s of free fall in the drop tower at the University of Bremen. Experiments demonstrate the physical principle behind the method of bubble formation and allow us to explore the dynamics of the resulting bubble jet after injection of the slug flow into a quiescent cavity in microgravity conditions as an efficient method of bubblespreadingandtransport.Detailed measurements of averagelocal velocities show thatbubblesare essentially passivewithrespecttothecarriermean flow,andtheinherentturbulenceofthe flowiscrucialforoptimalspreading of the bubbledistribution andreduction of bubble coalescence. Theshape of the bubblejet isstudied asa function of the Reynolds number. Finally, the degree of coalescence is also characterized and found to be remarkably small.

Preparation of suspensions of phospholipid-coated microbubbles by coaxial electrohydrodynamic atomization.

The use of phospholipid-coated microbubbles for medical applications is gaining considerable attention. However, the preparation of lipid-coated microbubble suspensions containing the ideal size and size distribution of bubbles still represents a considerable challenge. The most commonly used preparation methods of sonication and mechanical agitation result in the generation of polydisperse microbubbles with diameters ranging from less than 1 microm to greater than 50 microm. Efforts have been made via distinctly different techniques such as microfluidic and electrohydrodynamic bubbling to prepare lipid-coated microbubbles with diameters less than 10 microm and with a narrow size distribution, and recent results have been highly promising. In this paper, we describe a detailed investigation of the latter method that essentially combines liquid and air flow, and an applied electric field to generate microbubbles. A parametric plot was constructed between the air flow rate (Qg) and the lipid suspension flow rate (Ql) to identify suitable flow rate regimes for the preparation of phospholipid-coated microbubbles with a mean diameter of 6.6 microm and a standard deviation of 2.5 microm. The parametric plot has also helped in developing a scaling equation between the bubble diameter and the ratio Qg/Ql. At ambient temperature (22 degrees C), these bubbles were very stable with their size remaining almost unchanged for 160 min. The influence of higher temperatures such as the human body temperature (37 degrees C) on the size and stability of the microbubbles was also explored. It was found that the mean bubble diameter fell rapidly to begin with but then stabilized at 1-2 microm after 20 min.

Microbubble suspension as a carrier of oxygen and acclimated bacteria for phenanthrene biodegradation

The applicability of microbubble suspension made of saponin as a biodegradation-enhancing carrier where oxygen and pollutant-degraders are limited was studied. The saponin-microbubble suspension was used to deliver phenanthrene-degrading bacteria, inorganic nutrients, and oxygen. Bench-scale study was carried out to determine the physical properties of the microbubble suspension and to verify whether the delivered bacteria and oxygen were effectively used to degrade phenanthrene. A concentration of 2 g saponin/L H 2O generated stable microbubble suspension with a long half-drainage time and a high gas hold-up, and the addition of phenanthrene-degraders and inorganic salts to the saponin solution did not affect such properties. The flow of the microbubble suspension through a heterogeneous sand/clay-packed column occurred in two phases, with the liquid front advancing faster and the retarded gas front. The retarded gas front provided oxygen with bacteria, which enables phenanthrene biodegradation. Approximately 30% of the spiked phenanthrene was degraded in 21 days when one pore volume of 2.0 g/L saponin-microbubble suspension was applied whereas no phenanthrene decrease was observed following the application of the same saponin solution without microbubble generation. The decrease mainly occurred at the lower part of the column where the supply of oxygen by the microbubble was concentrated.

High-speed optical observations and simulation results of SonoVue microbubbles at low-pressure insonation

Abstract-Modified Rayleigh-Plesset models are commonly used to characterize the acoustic response of microbubbles under ultrasound exposure. In most instances these models have been parameterized through acoustic measurements taken from bulk suspensions of microbubbles. The aim of this study was to parameterize the Hoff model for the commercial contrast agent SonoVue using optically observed oscillations from individual microbubbles recorded with a high-speed camera. The shell elasticity model term was tuned to fit simulation data to the measured oscillations while the shell viscosity parameter was held constant at 1 Pa??s. The results demonstrate a limited ability of the model to predict the microbubble behavior. The shell elasticity parameter was found to vary proportionally between 10 and 80 MPa with the initial microbubble diameter, implying the viscoelastic shell terms are not a constant property of the shell material. Further analysis using a moving window optimization to probe the microbubble responses suggests that the elasticity of the shell can increase by up to 50% over the course of insonation, particularly for microbubbles oscillating nearer to their resonant frequency. Microbubble oscillations were modeled more successfully by incorporating a varying elasticity term into the model.

Dynamics of Bubble Formation in Highly Viscous Liquids

There has recently been considerable interest in the development of devices for the preparation of monodisperse microbubble suspensions for use as ultrasound contrast agents and drug delivery vehicles. These applications require not only a high degree of bubble uniformity but also a maximum bubble size of 8 mum, and this provides a strong motivation for developing an improved understanding of the process of bubble formation in a given device. The aim of this work was to investigate bubble formation in a T-junction device and determine the influence of the different processing parameters upon bubble size, in particular, liquid viscosity. Images of air bubble formation in a specially designed T-junction were recorded using a high-speed camera for different ratios of liquid to gas flow rate (Ql/Qg) and different liquid viscosities (microl). It was found that theoretical predictions of the flow profile in the focal region based on analysis of axisymmetric Stokes flow were accurate to within 6% when compared with the experimental data, indicating that this provided a suitable means of describing the bubble formation process. Both the theoretical and experimental results showed that Ql/Qg and mul had a significant influence upon bubble formation and eventual size, with higher flow rates and higher viscosities producing smaller bubbles. There were, however, found to be limiting values of Ql/Qg and mul beyond which no further reduction in bubble size was achieved.

Preparative-scale one-pot syntheses of hexafluoro-1,3-butadiene

Hexafluoro-1,3-butadiene, with its negligible global warming potential, is required in quantities for application in plasma dielectric etching in semiconductor industry and as gaseous microbubble suspension contrast agents in diagnostic ultrasound imaging. Three efficient protocols for the preparation of perfluoro-1,3-butadiene in 62–70% overall yields have been described. They involve the coupling of (1) iodotrifluoroethylene (ITFE) with activated copper, (2) trifluorovinylzinc bromide in the presence of copper (II) or iron (III) salts and (3) trifluorovinylzinc chloride, prepared from 1,1,1,2-tetrafluorethane (HFC 134a) in the presence of copper (II) or iron (III) salts.

Flow Characteristics of Microbubble Suspensions in Porous Media as an Oxygen Carrier

AbstractMicrobubble suspensions were generated as an oxygen carrier for aerobic biodegradation, and their flow characteristics in porous media were investigated. Commercial surfactants including sodium dodecyl sulfate (SDS), and dodecylethyldimethylammonium bromide (DEDAB), saponin (a natural surfactant), and collagen (a protein hydrolysate) were examined as base materials for generating microbubble suspensions. Among them, 2×CMC (critical micellar concentration) of SDS, DEDAB, and saponin developed microbubble suspensions with the highest gas hold‐up and half‐drainage time. Visualization of the flow patterns in sand showed that the microbubble suspensions were separated into a liquid and gas phase directly after injection, showing much faster movement of liquid phase flow. The gas front of the microbubble suspensions flowed in a plug‐flow manner, particularly in cases of SDS and DEDAB. The experimental results from both homogeneous and heterogeneous cells confirmed that the microbubble flow could overcome the heterogeneity in porous media. However, the plug‐flow characteristics and flow propagation of the microbubble suspensions to the low‐permeability zone was accompanied by a large pressure drop, which needs to be considered for future field application.

Phospholipids-based microbubbles sonoporation pore size and reseal of cell membrane cultured in vitro

Objective: To investigate phospholipids-based microbubbles induced sonoporation and cell membrane reseal in vitro under various conditions.Methods: A breast cancer cell line SK-BR-3 was used to investigate ultrasonic sonoporation under various conditions. Atomic force microscopy (AFM) scanning techniques were employed to observe the change of membrane pores.Results: Normal SK-BR-3 cells membrane pores were evenly distributed and less than 1 μm. After ultrasound exposure, membrane pores were enlarged at different degree depending on ultrasound exposure durations, filling gas species and microbubble suspension concentration. With microbubble suspension concentration being increased to 5% or ultrasound exposure reached 30 s, membrane pores in fluorocarbon (C3F8 or SF6)-filled microbubble groups exceeded 1 μm, which were significantly larger than that of air-filled microbubble group. Membrane pores were about 2–3 μm under ultrasound 60 s with 5% fluorocarbon-filled microbubble suspension. After 24 h of incubation, most of the enlarged membrane pores could reseal to normal size, which corresponded to cell viability.Conclusions: Membrane pores can be obviously enlarged by ultrasonic sonoporation of fluorocarbon-filled microbubbles, whose reseal time depended on ultrasound exposure duration and microbubble suspension concentration.

Novel methods for preparing phospholipid coated microbubbles

Two new methods for preparing phospholipid coated microbubble suspensions are elucidated. Firstly, co-axial electrohydrodynamic atomisation was utilized to generate 3-7 microm diameter microbubbles. Secondly, a specially designed and constructed T-junction device was used to prepare monodisperse microbubbles. Characteristics of microbubbles prepared by these two methods are compared with those obtained by sonication of the phospholipid suspension.