Acoustic Microstreaming Research Articles

A Novel Acoustic Modulation of Oscillating Thin Elastic Membrane for Enhanced Streaming in Microfluidics and Nanoscale Liposome Production.

Liposomes are widely utilized in therapeutic nanosystems as promising drug carriers for cancer treatment, which requires a meticulous synthesis approach to control the nanoprecipitation process. Acoustofluidic platforms offer a favorable synthesis environment by providing robust agitation and rapid mixing. Here, a novel high-throughput acoustofluidic micromixer is presented for a solvent and solvent-free synthesis of ultra-small and size-tunable liposomes. The size-tunability is achieved by incorporating glycerol as a new technique into the synthesis reagents, serving as a size regulator. The proposed device utilizes the synergistic effects of vibrating trapped microbubbles and an oscillating thin elastic membrane to generate vigorous acoustic microstreaming. The working principle and mixing mechanism of the device are explored numerically and experimentally. The platform exhibits remarkable mixing efficacy for aqueous and viscous solutions at flow rates up to 8000 µL/h, which makes it unique for high-throughput liposome formation and preventing aggregation. As a proof of concept, this study investigates the impact of phospholipid type and concentration, flow rate, and glycerol on the size and size distribution of liposomes. The results reveal a significant size reduction, from ≈900 nm to 40 nm, achieved by merely introducing 75% glycerol into the synthesis reagents, highlighting an innovative approach toward size-tunable liposomes.

Micromixer driven by bubble-induced acoustic microstreaming for multi-ink 3D bioprinting.

Recently, the 3D printing of cell-laden hydrogel structures, known as bioprinting, has received increasing attention owing to advances in tissue engineering and drug screening. However, a micromixing technology that efficiently mixes viscous bioinks under mild conditions is needed. Therefore, this study presents a novel method for achieving homogeneous mixing of multiple inks in 3D bioprinting through acoustic stimulation. This technique involves generating an acoustic microstream through bubble oscillations inside a 3D bioprinting nozzle. We determined the optimal hole design for trapping a bubble, hole arrangement, and voltage for efficient mixing, resulting in a four-fold increase in mixing efficiency compared to a single bubble arrangement. Subsequently, we propose a nozzle design for efficient mixing during bioprinting. The proposed nozzle design enabled the successful printing of line structures with a uniform mixture of different viscous bioinks, achieving a mixing efficiency of over 80% for mixing 0.5-1.0 wt% sodium alginate aqueous solutions. Additionally, acoustic stimulation had no adverse effects on cell viability, maintaining a high cell viability of 88% after extrusion. This study presents the first use of a bubble micromixer in 3D bioprinting, demonstrating gentle yet effective multi-ink mixing. We believe this approach will broaden 3D printing applications, particularly for constructing functional structures in 3D bioprinting.

Modeling stable cavitation of coated microbubbles: A framework integrating smoothed dissipative particle dynamics and the Rayleigh-Plesset equation.

Coated microbubbles are widely used in medical applications, particularly in enhanced drug and gene delivery. One of the mechanisms underlying these applications involves the shear stress exerted on the cell membrane by acoustic microstreaming generated through cavitation bubbles. In this study, we develop a novel simulation approach that combines the smooth dissipative particle dynamics (SDPD) simulation method with numerical modeling of the Rayleigh-Plesset-like equation in an ad hoc manner to simulate stable cavitation of microbubbles at microsecond and micrometer scales. Specifically, the SDPD method is utilized to model fluid dynamics, while the Rayleigh-Plesset-like equation is employed to describe bubble dynamics. Adopting a 1.5 μm coated microbubble driven by ultrasound with a frequency of 2MHz and a pressure of 500kPa as a representative example, we observe a high-velocity microstreaming pattern emerging around the bubble on a very small scale of a few micrometers after only a few microseconds. These spatiotemporal scales may pose challenges for experimental observation. The formation of this microstreaming arises from the opposing motion of the fluid layer next to the bubble and the fluid layers further away. Furthermore, our simulations reveal high shear stress levels of thousands of Pascals exerted on a wall located a few micrometers from the bubble. This contrasts with the shear stress values of a few Pascals calculated from theoretical models in the literature, which do not incorporate radial streaming into their theories. The implications of our results for bubble cavitation-induced pore formation on the cell membrane are discussed in some details.

Bioeffects of microbubbles in tissue engineering: Modeling collapsing jets and microstreaming

Ultrasound contrast agents are micron-sized bubbles coated with lipids/proteins to stabilize them in the bloodstream. Apart from enhancing the contrast of the image, they have been implicated in numerous harmful and beneficial bioeffects. I will present an overview of our research emphasizing recent efforts on microbubbles-based non-invasive pressure estimation and ultrasound-assisted bone and cartilage tissue engineering in 3D printed scaffolds. Low-intensity pulsed ultrasound (LIPUS) in conjunction with microbubbles has been shown in our lab to facilitate bone and cartilage formation from mesenchymal stem cells. We will discuss nonlinear shape oscillations of microbubbles and acoustic microstreaming that are responsible for such bioeffects. We studied them using boundary element (BEM) simulation and perturbative analysis of an encapsulated microbubble near a vessel wall. For the BEM solution, the coating of the microbubble is modeled as a viscoelastic interface using an in-house developed strain-softening model (exponential elasticity model). The influence of the shell model on the stability of the numerical simulation during the microbubble jet formation has been investigated. The effects of ultrasound excitation parameters and mechanical properties of the coating, i.e., shell viscosity and elasticity, on the bubble behaviors and the velocity and pressure in the surrounding fluid, have been studied.

Acoustic radiation force on a heated spherical particle in a fluid including scattering and microstreaming from a standing ultrasound wave.

Analytical expressions are derived for the time-averaged, quasisteady, acoustic radiation force on a heated, spherical, elastic, solid microparticle suspended in a fluid and located in an axisymmetric incident acoustic wave. The heating is assumed to be spherically symmetric, and the effects of particle vibrations, sound scattering, and acoustic microstreaming are included in the calculations of the acoustic radiation force. It is found that changes in the speed of sound of the fluid due to temperature gradients can significantly change the force on the particle, particularly through perturbations to the microstreaming pattern surrounding the particle. For some fluid-solid combinations, the effects of particle heating even reverse the direction of the force on the particle for a temperature increase at the particle surface as small as 1K.

Acoustic radiation force on a spherical thermoviscous particle in a thermoviscous fluid including scattering and microstreaming.

We derive general analytical expressions for the time-averaged acoustic radiation force on a small spherical particle suspended in a fluid and located in an axisymmetric incident acoustic wave. We treat the cases of the particle being either an elastic solid or a fluid particle. The effects of particle vibrations, acoustic scattering, acoustic microstreaming, heat conduction, and temperature-dependent fluid viscosity are all included in the theory. Acoustic streaming inside the particle is also taken into account for the case of a fluid particle. No restrictions are placed on the widths of the viscous and thermal boundary layers relative to the particle radius. We compare the resulting acoustic radiation force with that obtained from previous theories in the literature, and we identify limits, where the theories agree, and specific cases of particle and fluid materials, where qualitative or significant quantitative deviations between the theories arise.

The role of surface tension on the growth of bubbles by rectified diffusion

Rectified diffusion has wide and important applications in sonochemistry, ultrasonic cleaning and medical ultrasound. Recent experimental results have demonstrated that the addition of surfactant substantially enhances bubble growth rate. As a hypothesis, this was widely attributed to the acoustic microstreaming and mass transfer resistance caused by surfactants. In this research, the effects of the surfactant of sodium dodecyl sulphate on the rectification have been simulated by considering only the variation of the surface tension coefficient due to the surfactant. The computations are carried out using a newly developed tractable model based on the multi-scale method and the method of matched asymptotic expansions, which allows the prediction of bubble growth taking place over millions of oscillation cycles. The rate of bubble growth observed in the experiments is accurately predicted by our computations, for a range for bulk surfactant SDS concentrations less than or equal to 2.4 mM. Contrary to the widely held hypothesis in the published literature, this has demonstrated that the dominant physical mechanisms remain the shell and area effects in this range of bulk surfactant concentrations. The further enhancement of bubble growth rate provided by either acoustic microstreaming or the resistance to mass transfer is only evident at higher bulk surfactant concentrations. Therefore, the role of surface tension in rectified diffusion for aqueous surfactant solutions is more significant than previously understood. The new results also show that the bubble growth rate is sensitive to small changes in the bubble radius which may account for its unpredictability in applications of sonochemistry.

Ultrasonic-assisted removal of cationic and anionic dyes residues from wastewater using functionalized triptycene-based polymers of intrinsic microporosity (PIMs)

In this work, a series of hypercrosslinked polymers of intrinsic microporosity (HCP-PIMs), namely nitro-triptycene (TRIP-NO2), amino-triptycene (TRIP-NH2), sulfonated-triptycene (TRIP-SO3H) and hydrocarbon-triptycene (TRIP-HC), are employed for the adsorption of organic dyes from wastewater. The materials show the efficient removal of cationic (malachite green, MG) and anionic (methyl orange, MO) dyes. The adsorption parameters herein investigated include the initial pH, the adsorbate concentration and the contact time, with the aim to elucidate their effect on the adsorption process. Furthermore, the adsorption kinetic and isotherms are studied, and the findings suggest the results fit well with pseudo-second-order kinetics and Langmuir model. The reported maximum adsorption capacity is competitive for all the tested polymers. More specifically, TRIP-SO3H and TRIP-HC exhibit adsorptions of ~ 303 and ~ 270 mg g−1 for MG and MO, respectively. The selectivity toward cationic and anionic dyes is assessed by mixing the two dyes, and showing that TRIP-HC completely removes both species, whereas TRIP-NO2, TRIP-NH2 and TRIP-SO3H show an enhanced selectivity toward the cationic MG, compared to the anionic MO. The effect of the type of water is assessed by performing ultrasonic-assisted adsorption experiments, using TRIP-SO3H and TRIP-HC in the presence of either tap or seawater. The presence of competing ions and their concentrations is evaluated by ICP-MS. Our study shows that tap water does not have a detrimental effect on the adsorption of both polymers, whereas, in the presence of seawater, the performance of TRIP-HC toward MO proved to be more stable than MG with TRIP-SO3H, which is probably due to a larger concentration of competing ions. Comparison between ultrasonic-assisted and magnetic stirring adsorption demonstrates that the former exhibits a greater efficiency. This seems due to a more rapid mass transfer, driven by the formation of high velocity micro-jets, acoustic microstreaming and shock waves, at the polymer surface. Reusability studies show a good stability up to five adsorption–desorption cycles.

Water Skating Miniature Robot Propelled by Acoustic Bubbles.

This paper presents a miniature robot designed for monitoring its surroundings and exploring small and complex environments by skating on the surface of water. The robot is mainly made of extruded polystyrene insulation (XPS) and Teflon tubes and is propelled by acoustic bubble-induced microstreaming flows generated by gaseous bubbles trapped in the Teflon tubes. The robot's linear motion, velocity, and rotational motion are tested and measured at different frequencies and voltages. The results show that the propulsion velocity is proportional to the applied voltage but highly depends on the applied frequency. The maximum velocity occurs between the resonant frequencies for two bubbles trapped in Teflon tubes of different lengths. The robot's maneuvering capability is demonstrated by selective bubble excitation based on the concept of different resonant frequencies for bubbles of different volumes. The proposed water skating robot can perform linear propulsion, rotation, and 2D navigation on the water surface, making it suitable for exploring small and complex water environments.

Ultrasonic Steering Wheels: Turning Micromotors by Localized Acoustic Microstreaming.

The ability to steer micromotors in specific directions and at precise speeds is highly desired for their use in complex environments. However, a generic steering strategy that can be applied to micromotors of all types and surface coatings is yet to be developed. Here, we report that ultrasound of ∼100 kHz can spin a spherical micromotor so that it turns left or right when moving forward, or that it moves in full circles. The direction and angular speeds of their spinning and the radii of circular trajectories are precisely tunable by varying ultrasound voltages and frequencies, as well as particle properties such as its radius, materials, and coating thickness. Such spinning is hypothesized to originate from the circular microstreaming flows localized around a solid microsphere vibrating in ultrasound. In addition to causing a micromotor to spin, such streaming flows also helped release cargos from a micromotor during a capture-transport-release mission. Localized microstreaming does not depend on or interference with a specific propulsion mechanism and can steer a wide variety of micromotors. This work suggests that ultrasound can be used to steer microrobots in complex, biologically relevant environments as well as to steer microorganisms and cells.

Microfluidic viscometer by acoustic streaming transducers.

Measurement of fluid viscosity represents a huge need for many biomedical and materials processing applications. Sample fluids containing DNA, antibodies, protein-based drugs, and even cells have become important therapeutic options. The physical properties, including viscosity, of these biologics are critical factors in the optimization of the biomanufacturing processes and delivery of therapeutics to patients. Here we demonstrate an acoustic microstreaming platform termed as microfluidic viscometer by acoustic streaming transducers (μVAST) that induces fluid transport from second-order microstreaming to measure viscosity. Validation of our platform is achieved with different glycerol content mixtures to reflect different viscosities and shows that viscosity can be estimated based on the maximum speed of the second-order acoustic microstreaming. The μVAST platform requires only a small volume of fluid sample (∼1.2 μL), which is 16-30 times smaller than that of commercial viscometers. In addition, μVAST can be scaled up for ultra-high throughput measurements of viscosity. Here we demonstrate 16 samples within 3 seconds, which is an attractive feature for automating the process flows in drug development and materials manufacturing and production.

Sonoprinting nanoparticles on cellular spheroids via surface acoustic waves for enhanced nanotherapeutics delivery.

Nanotherapeutics, on their path to the target tissues, face numerous physicochemical hindrances that affect their therapeutic efficacy. Physical barriers become more pronounced in pathological tissues, such as solid tumors, where they limit the penetration of nanocarriers into deeper regions, thereby preventing the efficient delivery of drug cargo. To address this challenge, we introduce a novel approach that employs surface acoustic wave (SAW) technology to sonoprint and enhance the delivery of nanoparticles onto and into cell spheroids. Our SAW platform is designed to generate focused and unidirectional acoustic waves for creating vigorous acoustic streaming while promoting Bjerknes forces. The effect of SAW excitation on cell viability, as well as the accumulation and penetration of nanoparticles on human breast cancer (MCF 7) and mouse melanoma (YUMM 1.7) cell spheroids were investigated. The high frequency, low input voltage, and contact-free nature of the proposed SAW system ensured over 92% cell viability for both cell lines after SAW exposure. SAW sonoprinting enhanced the accumulation of 100 nm polystyrene particles on the periphery of the spheroids to near four-fold, while the penetration of nanoparticles into the core regions of the spheroids was improved up to three times. To demonstrate the effectiveness of our SAW platform on the efficacy of nanotherapeutics, the platform was used to deliver nanoliposomes encapsulated with the anti-cancer metal compound copper diethyldithiocarbamate (CuET) to MCF 7 and YUMM 1.7 cell spheroids. A three-fold increase in the cytotoxic activity of the drug was observed in spheroids under the effect of SAW, compared to controls. The capacity of SAW-based devices to be manufactured as minuscule wearable patches can offer highly controllable, localized, and continuous acoustic waves to enhance drug delivery efficiency to target tissues.

Development of a novel microfluidic biosensing platform integrating micropillar array electrode and acoustic microstreaming techniques

Quantifying biomarkers at the early stage of the disease is challenging due to the low abundance of biomarkers in the sample and the lack of sensitive techniques. This article reports the development of a novel microfluidic electrochemical biosensing platform to address this challenge. The electrochemical sensing is achieved by utilizing a micropillar array electrode (μAE) coated with 3D bimetallic Pt–Pd nanotrees to enhance the sensitivity. A bubble-based acoustic microstreaming technique is integrated with the device to increase the contact of analyte molecules with the surface of electrodes to further enhance the electrochemical performance. The current density of Pt–Pd NTs/μAE with acoustic microstreaming is nearly 22 times that of the bare planar electrode in potassium ferrocyanide solution. The developed biosensor has demonstrated excellent sensing performance. For hydrogen peroxide detection, both the Pt–Pd NTs/μAE and acoustic microstreaming contribute to the sensitivity enhancement. The current density of the Pt–Pd NTs/μAE is approximatively 28 times that of the bare μAE. With acoustic microstreaming, this enhancement is further increased by nearly 1.6 times. The platform has a linear detection range of 5–1000 μM with a LOD of 1.8 μM toward hydrogen peroxide detection, while for sarcosine detection, the linear range is between 5 and 100 μM and LOD is 2.2 μM, respectively. Furthermore, the sarcosine biosensing shows a high sensitivity of 667 μA mM−1∙cm−2. Such a sensing platform has the potential as a portable device for high sensitivity detection of biomarkers.

Influence of particle shape and material on the acoustic radiation force and microstreaming in a standing wave.

In view of its influence on the acoustic radiation force, we investigate the microstreaming around a small solid elastic particle in an ultrasonic standing wave in dependence of its material properties and shape. The configuration is axisymmetric, making it accessible to numerical methods, such as the finite element method. The results reveal a transition from viscous scattering- to microstreaming-dominated acoustic radiation force that depends on the particle density. When a deviation of the particle shape from a sphere becomes smaller than the viscous boundary layer thickness, we show that the influence of the shape on the viscous contributions to the acoustic radiation force diminishes, allowing the use of theoretical models for a spherical particle. However, extreme asymmetric shape perturbations, such as crowns with sharp edges, can give rise to noticeable viscous contributions for a dense particle that is larger than the viscous boundary layer thickness. We also introduce a hybrid analytical model for the acoustic radiation force on a spherical particle that accounts for the microstreaming and particle compressibility and shows a good agreement with numerical simulations for an arbitrary particle size and density.

Acoustofluidics for simultaneous nanoparticle-based drug loading and exosome encapsulation

Nanocarrier and exosome encapsulation has been found to significantly increase the efficacy of targeted drug delivery while also minimizing unwanted side effects. However, the development of exosome-encapsulated drug nanocarriers is limited by low drug loading efficiencies and/or complex, time-consuming drug loading processes. Herein, we have developed an acoustofluidic device that simultaneously performs both drug loading and exosome encapsulation. By synergistically leveraging the acoustic radiation force, acoustic microstreaming, and shear stresses in a rotating droplet, the concentration, and fusion of exosomes, drugs, and porous silica nanoparticles is achieved. The final product consists of drug-loaded silica nanocarriers that are encased within an exosomal membrane. The drug loading efficiency is significantly improved, with nearly 30% of the free drug (e.g., doxorubicin) molecules loaded into the nanocarriers. Furthermore, this acoustofluidic drug loading system circumvents the need for complex chemical modification, allowing drug loading and encapsulation to be completed within a matter of minutes. These exosome-encapsulated nanocarriers exhibit excellent efficiency in intracellular transport and are capable of significantly inhibiting tumor cell proliferation. By utilizing physical forces to rapidly generate hybrid nanocarriers, this acoustofluidic drug loading platform wields the potential to significantly impact innovation in both drug delivery research and applications.

A theoretical model for the growth of spherical bubbles by rectified diffusion

Rectified diffusion is a bubble growth phenomenon that occurs in acoustic fields. Despite the existence of a well-established spherically symmetric mathematical model, theoretical results have been unsuccessful in reproducing the bubble growth even in the case of a single spherical bubble in the bulk. In the latter case, the influence of surfactants and acoustic microstreaming have been speculated as the explanation for this disagreement. In this article, an exact solution for the leading-order concentration of gas in the liquid is determined. Using this exact solution, the well-established mathematical model is reduced to a system of two ordinary differential equations for the spherical bubble radius and the mass of gas in the bubble. This simplified model predicts the rapid bubble growth observed in experiments of a single spherical bubble in the bulk. The new results show that the bubble growth is asymptotically larger than at later times when the mass flux is limited by the slow diffusion of gas in a much larger region of the liquid surrounding the bubble. The new results also show that this bubble growth is relatively insensitive to a reduction in surface tension.

Nonspherical dynamics and microstreaming of a wall-attached microbubble

Acoustic microstreaming is a nonlinear response of a fluid that undergoes high-amplitude acoustic stimulation and tends to viscously absorb it. The present experimental study investigates the generation of acoustic microstreaming induced by an oscillating wall-attached bubble undergoing nonspherical shape modes. From a microscope top view, the formation of particular flow signatures is explored for the main classes of spherical harmonics $Y_{nm}(\theta, \phi )$ : zonal ( $m = 0 < n$ ), sectoral ( $n = m > 0$ ) and tesseral ( $0 < m < n$ ) oscillation. The microstreaming induced by a bubble animated by a sectoral mode alone reveals a pattern characterized by a $4n$ -lobe flower shape. Tesseral modes give rise to 4m-lobe flower-shaped patterns. Finally, when sectoral and zonal modes coexist, two kinds of pattern stand out: $2n$ -lobe flower shape and $n$ -pointed star shape. The preferential emergence of one or another streaming pattern is discussed on the basis of the amplitude and phase shift between both shape modes.

Ultrasound enhancing the mass transfer of droplet microreactor for the synthesis of AgInS2 nanocrystals

Droplet microreactors are powerful tools for synthesizing chemicals, nano materials and biomolecules, but traditional methods, such as mechanical stirring, are hard to enhance the internal mass transfer of droplet microreactor due to its small reaction space. In this study, we propose a novel ultrasonic-enhancing method for droplet microreactor, and report the effect of bubbles inside micro-droplets for the first time. On this basis, the synthesis of AgInS2 nanocrystals is selected as model reaction to study the enhancement mechanism. It is discovered that the flow field in micro-droplet is significantly improved owing to acoustic streaming and cavitation microstreaming. Benefiting from them, the mass transfer coefficient is enhanced by 72.5 %, and AgInS2 nanocrystals are successfully synthesized with smaller, more uniform particle size, as well as more adjustable performance compared with no ultrasonic radiation condition. This work provides a new method and a deep insight of ultrasound enhancing droplet microreactor, enabling the continuous production of fine chemical synthesis.

Bubble-enhanced ultrasonic microfluidic chip for rapid DNA fragmentation.

DNA fragmentation is an essential process in developing genetic sequencing strategies, genetic research, as well as for the diagnosis of diseases with a genetic signature like cancer. Efficient on-chip DNA fragmentation protocols would be beneficial to process integration and open new opportunities for microfluidics in genetic applications. Here we present an acoustic microfluidic chip comprising an array of ultrasound-actuated microbubbles located at dedicated positions adjacent to a channel containing the DNA sample solution. The efficiency of the on-chip DNA fragmentation process arises mainly from tensile forces generated by acoustic streaming near the oscillating bubble interfaces, as well as a synergistic effect of streaming stress and ultrasonic cavitation. Acoustic microstreaming and the pressure distribution in the DNA channel were assessed by finite element simulation. We characterized the bubble-enhanced effect by measuring gene fragment size distributions with respect to different ultrasound parameters. For optimized on-chip conditions, purified lambda (λ) DNA (48.5 kbp) could be disrupted to fragments with an average size of 2 kbp after 30 s and down to 300 bp after 90 s. Mouse genomic DNA (1.4 kbp) fragmentation size decreased to 500 bp in 30 s and reduced further to 250 bp in 90 s. Bubble-induced fragmentation was more than 3 times faster than without bubbles. On-chip performance and process yield were found to be comparable to a sophisticated high-end commercial system. In this view, our new bubble-enhanced microfluidic approach is a promising tool for current and next generation sequencing platforms with high efficiency and good capacity. Moreover, the availability of an efficient on-chip DNA fragmentation process opens perspectives for implementing full molecular protocols on a single microfluidic platform.

Acoustic microstreaming produced by two interacting gas bubbles undergoing axisymmetric shape oscillations

An analytical theory is developed that describes acoustic microstreaming produced by two interacting bubbles. The bubbles are assumed to undergo axisymmetric oscillation modes, which can include radial oscillations, translation and shape modes. Analytical solutions are derived in terms of complex amplitudes of oscillation modes, which means that the modal amplitudes are assumed to be known and serve as input data when the velocity field of acoustic microstreaming is calculated. No restrictions are imposed on the ratio of the bubble radii to the viscous penetration depth and the distance between the bubbles. The interaction between the bubbles is considered both when the linear velocity field is calculated and when the second-order velocity field of acoustic microstreaming is calculated. Capabilities of the analytical theory are illustrated by computational examples.