Acoustic-holography-patterned primary hepatocytes possess liver functions.

Acoustic holography (AH), a promising approach for cell patterning, emerges as a powerful tool for constructing novel invitro 3D models that mimic organs and cancers features. However, understanding changes in cell function post-AH remains limited. Furthermore, replicating complex physiological and pathological processes solely with cell lines proves challenging. Here, we employed acoustical holographic lattice to assemble primary hepatocytes directly isolated from mice into a cell cluster matrix to construct a liver-shaped tissue sample. For the first time, we evaluated the liver functions of AH-patterned primary hepatocytes. The patterned model exhibited large numbers of self-assembled spheroids and superior multifarious core hepatocyte functions compared to cells in 2D and traditional 3D culture models. AH offers a robust protocol for long-term in vitro culture of primary cells, underscoring its potential for future applications in disease pathogenesis research, drug testing, and organ replacement therapy.

Enhanced Ultrasound Transmission through Skull Using Flexible Matching Layer with Gradual Acoustic Impedance.

Transcranial ultrasound imaging and therapy have gained significant attention due to their noninvasive nature, absence of ionizing radiation, and portability. However, the presence of the skull, which has a high acoustic impedance, presents a challenge for the penetration of ultrasound into intracranial tissue. This leads to a low transmission of ultrasound through the skull, hindering energy focusing and imaging quality. To address this challenge, we propose a novel approach that utilizes a flexible matching layer with gradual acoustic impedance to enhance ultrasound transmission through the skull. This matching layer is constructed using Poly(dimethylsiloxane) (PDMS)/tungsten powders as the structural component responsible for the gradual impedance, while agarose serves as the flexible matrix. Our simulation and experimental results demonstrate that the matching layer with an exponential gradual acoustic impedance significantly improves the ultrasound transmission coefficient across a wide frequency range compared to traditional quarter wavelength matching layers. Specifically, at 2 MHz, the maximum transmission coefficient reaches 49.5%, more than four times higher than that of the skull without a matching layer (only 11.7%). Additionally, the good flexibility of our matching layer ensures excellent adhesion to the curved surface of the skull, further enhancing its application potential in transcranial ultrasound imaging and therapy. The improved transmission performance allows for a lower ultrasound transmission power, effectively addressing overheating and safety issues.

Multi-frequency acoustic hologram generation with a physics-enhanced deep neural network.

Here, a physics-enhanced multi-frequency acoustic hologram deep neural network (PhysNet_MFAH) method is proposed for designing multi-frequency acoustic holograms, which is built by incorporating multiple physical models that represent the physical processes of acoustic waves propagation for a set of design frequencies into a deep neural network. It is demonstrated that one needs only to feed a set of frequency-specific target patterns into the network, the proposed PhysNet_MFAH method can automatically, accurately, and rapidly generate a high-quality multi-frequency acoustic hologram for holographic rendering of different target acoustic fields in the same or distinct regions of the target plane when driven at different frequencies. Remarkably, it is also demonstrated that the proposed PhysNet_MFAH method can achieve a higher quality of the reconstructed acoustic intensity fields than the existing optimization methods IASA and DS for designing multi-frequency acoustic holograms at a relatively fast-computational speed. Furthermore, the performance dependencies of the proposed PhysNet_MFAH method on different design parameters are established, which provide insight into the performance of the reconstructed acoustic intensity fields when subject to different design conditions of the proposed PhysNet_MFAH method. We believe that the proposed PhysNet_MFAH method can facilitate many potential applications of acoustic holograms, ranging from dynamic particle manipulation to volumetric display.

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.

Remote control of the recruitment and capture of endogenous stem cells by ultrasound for in situ repair of bone defects.

Stem cell-based tissue engineering has provided a promising platform for repairing of bone defects. However, the use of exogenous bone marrow mesenchymal stem cells (BMSCs) still faces many challenges such as limited sources and potential risks. It is important to develop new approach to effectively recruit endogenous BMSCs and capture them for in situ bone regeneration. Here, we designed an acoustically responsive scaffold (ARS) and embedded it into SDF-1/BMP-2 loaded hydrogel to obtain biomimetic hydrogel scaffold complexes (BSC). The SDF-1/BMP-2 cytokines can be released on demand from the BSC implanted into the defected bone via pulsed ultrasound (p-US) irradiation at optimized acoustic parameters, recruiting the endogenous BMSCs to the bone defected or BSC site. Accompanied by the daily p-US irradiation for 14 days, the alginate hydrogel was degraded, resulting in the exposure of ARS to these recruited host stem cells. Then another set of sinusoidal continuous wave ultrasound (s-US) irradiation was applied to excite the ARS intrinsic resonance, forming highly localized acoustic field around its surface and generating enhanced acoustic trapping force, by which these recruited endogenous stem cells would be captured on the scaffold, greatly promoting them to adhesively grow for in situ bone tissue regeneration. Our study provides a novel and effective strategy for in situ bone defect repairing through acoustically manipulating endogenous BMSCs.

Acoustic radiation force dependence on properties of elastic spherical shells in standing waves.

In this study, we theoretically investigate the acoustic radiation force acting on elastic spherical shells in standing waves in the dimensionless frequency range of 0

Clinical efficacy of different shoulder joint drug injections for rotator cuff injuries: A network meta-analysis.

BACKGROUND: Rotator cuff injury is a common clinical disorder of the musculoskeletal system. It is one of the most common causes of shoulder pain and shoulder joint dysfunction. Intra-articular injection therapy is one of the common conservative treatments for rotator cuff injury in clinical practice, but scholars have been controversial about the different drugs used for injection therapy and their clinical effectiveness, and there is no complete agreement on the choice of different injectables, and the existing original studies are mostly direct comparisons in 1 arm, lacking indirect comparisons, making it difficult to have a clear and comprehensive understanding. OBJECTIVE: A network Meta-analysis was used to comprehensively compare the effectiveness of drug injection therapies for rotator cuff injuries. METHODS: The PubMed, The Cochrane Library, EMbase, Chinese national knowledge infrastructure, Chinese BioMedical Literature on disc, China Science and Technology Journal Database, and Wan-Fang databases were searched for randomized controlled trials on the effectiveness of steroid injections for the comparative treatment of rotator cuff injury for the period April 19, 2013 to April 19, 2022 (the last decade). Two investigators independently screened the literature, extracted data according to inclusion and exclusion criteria, and evaluated the quality of the literature in parallel. Statistical analysis was performed using Stata 16.0 software to compare the differences in efficacy of each treatment measure and rank the efficacy using the ratio and 95% confidence interval (CI) as the effect indicator. RESULTS: 10 RCTs with a total of 861 patients with rotator cuff injury were included, involving 4 therapeutic measures: corticosteroid injection therapy (COR), platelet-rich plasma injection therapy (PRP), Hyaluronic acid injection therapy (HA), and prolotherapy therapy (PRO).Meta-analysis results showed that the ranking results of the 4 therapeutic measures were: corticosteroid injection + hyaluronic acid injection > platelet-rich plasma injection + corticosteroid injection > corticosteroid injection > platelet-rich plasma injection > PRO > platelet-rich plasma injection + hyaluronic acid injection > hyaluronic acid injection. CONCLUSION: we recommend that corticosteroid injections combined with hyaluronic acid injections can be used for the non-surgical conservative clinical management of rotator cuff injuries.

Brain Delivery of Curcumin Through Low-Intensity Ultrasound-Induced Blood-Brain Barrier Opening via Lipid-PLGA Nanobubbles.

BACKGROUND: Parkinson's disease (PD) is a progressive neurodegenerative disorder. Owing to the presence of blood-brain barrier (BBB), conventional pharmaceutical agents are difficult to the diseased nuclei and exert their action to inhibit or delay the progress of PD. Recent literatures have demonstrated that curcumin shows the great potential to treat PD. However, its applications are still difficult in vivo due to its poor druggability and low bioavailability through the BBB. METHODS: Melt-crystallization methods were used to improve the solubility of curcumin, and curcumin-loaded lipid-PLGA nanobubbles (Cur-NBs) were fabricated through encapsulating the curcumin into the cavity of lipid-PLGA nanobubbles. The bubble size, zeta potentials, ultrasound imaging capability and drug encapsulation efficiency of the Cur-NBs were characterized by a series of analytical methods. Low-intensity focused ultrasound (LIFU) combined with Cur-NB was used to open the BBB to facilitate curcumin delivery into the deep brain of PD mice, followed by behavioral evaluation for the treatment efficacy. RESULTS: The solubility of curcumin was improved by melt-crystallization methods, with 2627-fold higher than pure curcumin. The resulting Cur-NBs have a nanoscale size about 400 nm and show excellent contrast imaging performance. Curcumin drugs encapsulated into Cur-NBs could be effectively released when Cur-NBs were irradiated by LIFU at the optimized acoustic pressure, achieving 30% cumulative release rate within 6 h. Importantly, Cur-NBs combined with LIFU can open the BBB and locally deliver the curcumin into the deep-seated brain nuclei, significantly enhancing efficacy of curcumin in the Parkinson C57BL/6J mice model in comparison with only Cur-NBs and LIFU groups. CONCLUSION: In this work, we greatly improved the solubility of curcumin and developed Cur-NBs for brain delivery of curcumin against PD through combining with LIFU-mediating BBB. Cur-NBs provide a platform for these potential drugs which are difficult to cross the BBB to treat PD disease or other central nervous system (CNS) diseases.

Three-dimensional spiral motion of microparticles by a binary-phase logarithmic-spiral zone plate.

Acoustic vortex beams, which have both linear and angular momentum, can be used to make precise acoustic tweezers. Limited by the symmetry of a normal vortex beam, these tweezers are usually used for trapping or rotating particles in two dimensions. Here, the three-dimensional spiral motion of two soft particles of different sizes was realized using a vortex beam with a twisted focus, which was synthesized by a silicone binary-phase logarithmic-spiral zone plate. Numerical simulations and experimental measurements demonstrated that the beam had anisotropic focuses of crescent transverse intensity profiles and a screw phase dislocation with a singularity at the center. Experiments showed that a small particle (k0r ≈ 1.3) can follow the twisted intensity of the beam, but a large particle (k0r ≈ 4.7) spirals up away from the twisted field pattern. This is attributed to the dominant gradient force for the small particle, whereas the scattering effect induced a scattering force combined with a gradient force for the large particle. This focused twisted beam, which was generated with a structured silicone plate, and the three-dimensional spiral motion of microparticles, advance the development of simple, compact, and disposable acoustic devices for the precise and diverse manipulation of microparticles.

Targeted Delivery of Liposomal Temozolomide Enhanced Anti-Glioblastoma Efficacy through Ultrasound-Mediated Blood-Brain Barrier Opening.

Glioblastoma (GBM) is the commonest form of primary brain tumor in the central nervous system, with median survival below 15 months and only a 25% two-year survival rate for patients. One of the major clinical challenges in treating GBM is the presence of the blood-brain barrier (BBB), which greatly limits the availability of therapeutic drugs to the tumor. Ultrasound-mediated BBB opening provides a promising approach to help deliver drugs to brain tumors. The use of temozolomide (TMZ) in the clinical treatment of GBM has been shown to be able to increase survival in patients with GBM, but this improvement is still trivial. In this study, we developed a liposomal temozolomide formulation (TMZ-lipo) and locally delivered these nanoparticles into GBM through ultrasound-mediated BBB opening technology, significantly suppressing tumor growth and prolonging tumor-bearing animal survival. No significant side effects were observed in comparison with control rats. Our study provides a novel strategy to improve the efficacy of TMZ against GBM.

Early Detection and Reversal of Cell Apoptosis Induced by Focused Ultrasound-Mediated Blood-Brain Barrier Opening.

Focused ultrasound (FUS) combined with microbubbles (MBs) has recently emerged as a potential approach to open the blood-brain barrier (BBB) for delivering drugs into the brain. However, appropriate approaches are still lacking to monitor the sublethal damage during FUS-mediated BBB opening in vivo, especially the early stage cell apoptotic events. Here, we developed a kind of nanoprobe-loaded MBs (AV-ICG-NPs@MBs) which can monitor the apoptotic cells that occur during FUS-mediated BBB opening through encapsulating the annexin V-targeted nanoprobes AV-ICG-NPs into the cavity of lipid-PLGA hybrid MBs. When irradiated by FUS, AV-ICG-NPs@MBs in the cerebral blood vessels would produce cavitation, favoring the BBB opening. Meanwhile, AV-ICG-NPs@MBs would be destroyed and release their AV-ICG-NPs payload. These released AV-ICG-NPs can be further delivered into the brain via the destructed BBB and bind with the phosphatidylserine externalized on the membrane of apoptotic cells if this occurs, leading to the prolonged detention of fluorescent signals in the brain. Furthermore, we also provided an effective strategy to inhibit or reverse the possible damage to the brain from a FUS-mediated BBB opening technology, through developing AV-ICG-NPs/GAS@MBs that encapsulate the antioxidant gastrodin (GAS) into AV-ICG-NPs@MBs. Accompanied by FUS irradiation and bubble cavitation, GAS was released and delivered into the brain, where they scavenged the oxygen free radicals produced from cavitation, leading to significantly lower fluorescence signals in the brain due to the absence of externalized phosphatidylserine. In conclusion, our study provides an approach to monitor and inhibit cell apoptotic events during FUS-mediated BBB opening.

3-D Acoustic Tweezers Using a 2-D Matrix Array With Time-Multiplexed Traps.

The use of acoustic tweezers for precise manipulation of microparticles in the aqueous environment is essential and challenging for biomechanical applications in vivo. A 3-D acoustic tweezer is developed in this study for 3-D manipulation by using a two-dimensional (2-D) phased array consisting of 256 elements operating at 1.04 MHz. The emission phases of each element are iteratively determined by a backpropagation algorithm to generate multiple acoustic traps. Different traps are multiplexed in time, thus forming synthesized acoustic fields. We demonstrate the 3-D levitation and translation of positive acoustic contrast particles, a major class of bioparticles, in water by different acoustic traps, and compare the positional deviation along the intended path via experimentally measured trajectories. Improved manipulating stability was achieved by multiplexed acoustic traps. The 3-D acoustic tweezers proposed in this study provide a versatile approach of contactless bioparticle trapping and translation, paving the way toward future application of nanodroplet and microbubble manipulations.

A deep learning approach for the fast generation of acoustic holograms.

Acoustic holographic techniques are crucial in diverse applications, such as three-dimensional holographic display and particle manipulation. However, conventional methods for computer-generated acoustics holography rely heavily on iterative optimization algorithms, which are time-consuming and particularly hinder their capacity of generating a dynamic hologram in real time. Here, a deep learning approach based on U-Net is proposed to rapidly generate an acoustic hologram with optimal amplitude and phase maps. It is demonstrated that, after being trained with adequate data that are numerically synthesized by the pseudo-inverse method, the proposed deep learning approach can generate both amplitude and phase maps for new target images with an improved overall reconstruction quality. Remarkably, after the offline cost is compensated by a lower online cost for the proposed DL approach, the hologram generation speed is significantly accelerated by the proposed deep learning approach as compared with the pseudo-inverse method, especially for complicated or dynamic images. With the hierarchical feature learning capability and the fast online computational speed, the proposed deep learning approach can serve as a smart platform for rapidly generating complete maps of holograms for the sophisticated or dynamical target images, leading to the new possibility of real-time acoustic-hologram-based applications.

Self-Navigated 3D Acoustic Tweezers in Complex Media Based on Time Reversal.

Acoustic tweezers have great application prospects because they allow noncontact and noninvasive manipulation of microparticles in a wide range of media. However, the nontransparency and heterogeneity of media in practical applications complicate particle trapping and manipulation. In this study, we designed a 1.04 MHz 256-element 2D matrix array for 3D acoustic tweezers to guide and monitor the entire process using real-time 3D ultrasonic images, thereby enabling acoustic manipulation in nontransparent media. Furthermore, we successfully performed dynamic 3D manipulations on multiple microparticles using multifoci and vortex traps. We achieved 3D particle manipulation in heterogeneous media (through resin baffle and ex vivo macaque and human skulls) by introducing a method based on the time reversal principle to correct the phase and amplitude distortions of the acoustic waves. Our results suggest cutting-edge applications of acoustic tweezers such as acoustical drug delivery, controlled micromachine transfer, and precise treatment.

An Acoustic Meta-Skin Insulator.

Acoustic metamaterials with artificial microstructures are attractive to realize intriguing functions, including efficient waveguiding, which requires large impedance mismatches to realize total side reflection with negligible transmission and absorption. While large impedance mismatch can be readily realized in an air environment, acoustic waveguiding in an underwater environment remains elusive due to insufficient impedance mismatch of state-of-the-art metamaterials. Here, a superhydrophobic acoustic metasurface of microstructured poly(vinylidene fluoride) membrane, referred to as a "meta-skin" insulator, which is able to confine acoustic waves in an all-angle and wide spectrum range due to tremendous impedance mismatch at stable air/water interfaces, viz., the Cassie-Baxter state is demonstrated. By utilizing the meta-skin insulator with broadband and high throughput, orbital-angular-momentum multiplexing at a high spectral efficiency and binary coding along large-angle bending channels for bit-error-free acoustic data transmission in an underwater environment are demonstrated. Very different from optical and/or electrical cable communications, acoustic waves can be simply and effectively coupled into remote meta-skin acoustic fibers from free space, which is technologically significant for long-haul and anti-interference communication. This work can enlighten many fluidic applications based on efficient waveguiding, such as in vivo ultrasound medical treatment and imaging.

Ultrasonic microstreaming for complex-trajectory transport and rotation of single particles and cells.

Precisely controllable transport and rotation of microparticles and cells has great potential to enable new capabilities for single-cell level analysis. In this work, we present versatile ultrasonic microstreaming based manipulation that enables active and precise control of transport and rotation of individual microscale particles and biological cells in a microfluidic device. Two different types of ultrasonic microstreaming flow patterns can be produced by oscillating embedded microstructures in circular and rectilinear vibration modes, which have been validated by both numerical simulation and experimental observation. We have further showcased the ability to transport individual microparticles along the outlines of complex alphabet letters, demonstrating the versatility and simplicity of single-particle level manipulation with bulk vibration.

Development of Scalable 2D Plane Array for Transcranial Ultrasonic Neuromodulation on Non-Human Primates: An Ex Vivo Study.

Ultrasonic neuromodulation on large animals, like non-human primates, requires the array transducer to have a good steering ability to arbitrarily stimulate various brain locations. Moreover, due to the different sizes of the animal heads, the array is preferred to be conveniently adjustable to different aperture sizes. To meet these requirements, a scalable 2D plane array, which can consist of up to tens of 256-element square modules, has been designed and fabricated in this study. Arrays with large apertures, consisting of 12 and 48 modules (including 3072 and 12288 elements, respectively), have been assembled and tested. The square modules were driven at a center frequency of 1.04 MHz. The transcranial and steering abilities of the modules were verified in a water tank on an ex vivo macaque skull. A focused beam transmitted by single module could pass through the skull with its peak acoustic pressure reduced to 18.1% of that in the free-field, with an acceptable transverse steering range up to 10 mm. Moreover, a much smaller focal spot was obtained by simultaneous transmission using two vertical-placed modules. Compared to the single module, the focal zone axial size was reduced from 29.7 mm to 3.3 mm, and the "stripes" pattern in the focal zone caused by standing waves was eliminated using a difference-frequency transmitting strategy. Such scalable 2D plane array, which can provide a small-size focal spot and has a large steering range and an acceptable transcranial ability, can be useful in research on ultrasonic neuromodulation on non-human primates.

Sonoporation of Cells by a Parallel Stable Cavitation Microbubble Array.

Sonoporation is a targeted drug delivery technique that employs cavitation microbubbles to generate transient pores in the cell membrane, allowing foreign substances to enter cells by passing through the pores. Due to the broad size distribution of microbubbles, cavitation events appear to be a random process, making it difficult to achieve controllable and efficient sonoporation. In this work a technique is reported using a microfluidic device that enables in parallel modulation of membrane permeability by an oscillating microbubble array. Multirectangular channels of uniform size are created at the sidewall to generate an array of monodispersed microbubbles, which oscillate with almost the same amplitude and resonant frequency, ensuring homogeneous sonoporation with high efficacy. Stable harmonic and high harmonic signals emitted by individual oscillating microbubbles are detected by a laser Doppler vibrometer, which indicates stable cavitation occurred. Under the influence of the acoustic radiation forces induced by the oscillating microbubble, single cells can be trapped at an oscillating microbubble surface. The sonoporation of single cells is directly influenced by the individual oscillating microbubble. The parallel sonoporation of multiple cells is achieved with an efficiency of 96.6 ± 1.74% at an acoustic pressure as low as 41.7 kPa.

Selective photothermal ablation of cancer cells by patterned gold nanocages using surface acoustic waves.

The patterning of nanoparticles, which are promising photothermal agents, is of great importance to selectively and precisely ablate tissues by thermal effects. In this paper, we demonstrated that nano-sized gold particles (gold nanocages, AuNCS) with a hollow structure could be used to generate various wavefront patterns of surface acoustic waves (SAWs) and the aligned AuNC lines facilitated the destruction of cancer cells by the thermal effect with high spatial resolution. The hollow structure improved the acoustic sensitivity of AuNCs, making them more sensitive to the acoustic radiation force. Moreover, the multi-scale patterning of AuNCs could be achieved by the interference of multiple acoustic beams. Given the photothermal characteristics of AuNCs, selective temperature elevation within a micrometer-sized region could be realized when the patterned AuNCs were irradiated by a laser. The cancer cells where the patterned AuNCs were located were eliminated by thermal ablation, while other cells remained alive. In particular, the acoustic frequency used in this study was as low as 11. 35 MHz and was in the range of diagnostic ultrasound (less than 12 MHz), offering a potential to serve as a powerful tool in clinical applications.

Ultrasonic super-oscillation wave-packets with an acoustic meta-lens.

The Schrödinger equation is a fundamental equation to describe the wave function of a quantum-mechanical system. The similar forms between the Schrödinger equation and the paraxial wave equation allow a paradigm shift from the quantum mechanics to classical fields, opening up a plethora of interesting phenomena including the optical super-oscillatory behavior. Here, we propose an ultrasonic meta-lens for generating super-oscillation acoustic wave-packets with different spatial momenta and then superimposing them to a diffraction-limit-broken spot, visually represented by the ring-shaped trapping of tiny particles. Moreover, based on the focused super-oscillation packets, we experimentally verify proof-of-concept super-resolution ultrasound imaging, opening up the arena of super-oscillation ultrasonics for advanced acoustic imaging, biomedical applications, and versatile far-field ultrasound control.