Ultrasound-responsive microbubbles and nanodroplets: A pathway to targeted drug delivery.

Ultrasound-responsive agents have shown great potential as targeted drug delivery agents, effectively augmenting cell permeability and facilitating drug absorption. This review focuses on two specific agents, microbubbles and nanodroplets, and provides a sequential overview of their drug delivery process. Particular emphasis is given to the mechanical response of the agents under ultrasound, and the subsequent physical and biological effects on the cells. Finally, the state-of-the-art in their pre-clinical and clinical implementation are discussed. Throughout the review, major challenges that need to be overcome in order to accelerate their clinical translation are highlighted.

Cavitation cloud formation and surface damage of a model stone in a high-intensity focused ultrasound field.

This work investigates the fundamental role of cavitation bubble clouds in stone comminution by focused ultrasound. The fragmentation of stones by ultrasound has applications in medical lithotripsy for the comminution of kidney stones or gall stones, where their fragmentation is believed to result from the high acoustic wave energy as well as the formation of cavitation. Cavitation is known to contribute to erosion and to cause damage away from the target, yet the exact contribution and mechanisms of cavitation remain currently unclear. Based on in situ experimental observations, post-exposure microtomography and acoustic simulations, the present work sheds light on the fundamental role of cavitation bubbles in the stone surface fragmentation by correlating the detected damage to the observed bubble activity. Our results show that not all clouds erode the stone, but only those located in preferential nucleation sites whose locations are herein examined. Furthermore, quantitative characterizations of the bubble clouds and their trajectories within the ultrasonic field are discussed. These include experiments with and without the presence of a model stone in the acoustic path length. Finally, the optimal stone-to-source distance maximizing the cavitation-induced surface damage area has been determined. Assuming the pressure magnitude within the focal region to exceed the cavitation pressure threshold, this location does not correspond to the acoustic focus, where the pressure is maximal, but rather to the region where the acoustic beam and thereby the acoustic cavitation activity near the stone surface is the widest.

Shell viscosity estimation of lipid-coated microbubbles.

Understanding the shell rheology of ultrasound contrast agent microbubbles is vital for anticipating their bioeffects in clinical practice. Past studies using sophisticated acoustic and optical techniques have made enormous progress in this direction, enabling the development of shell models that adequately reproduce the nonlinear behaviour of the coated microbubble under acoustic excitation. However, there have also been puzzling discrepancies and missing physical explanations for the dependency of shell viscosity on the equilibrium bubble radius, which demands further experimental investigations. In this study, we aim to unravel the cause of such behaviour by performing a refined characterisation of the shell viscosity. We use ultra-high-speed microscopy imaging, optical trapping and wide-field fluorescence to accurately record the individual microbubble response upon ultrasound driving across a range of bubble sizes. An advanced model of bubble dynamics is validated and employed to infer the shell viscosity of single bubbles from their radial time evolution. The resulting values reveal a prominent variability of the shell viscosity of about an order of magnitude and no dependency on the bubble size, which is contrary to previous studies. We find that the method called bubble spectroscopy, which has been used extensively in the past to determine the shell viscosity, is highly sensitive to methodology inaccuracies, and we demonstrate through analytical arguments that the previously reported unphysical trends are an artifact of these biases. We also show the importance of correct bubble sizing, as errors in this aspect can also lead to unphysical trends in shell viscosity, when estimated through a nonlinear fitting from the time response of the bubble.

Synchrotron X-ray imaging of the onset of ultrasonic horn cavitation.

High-power ultrasonic horns operating at low frequency are known to generate a cone-shaped cavitation bubble cloud beneath them. The exact physical processes resulting in the conical structure are still unclear mainly due to challenges associated with their visualization. Herein, we address the onset of the cavitation cloud by exploiting high-speed X-ray phase contrast imaging. It reveals that the cone formation is not immediate but results from a three-step phenomenology: (i) inception and oscillation of single bubbles, (ii) individual cloud formation under splitting or lens effects, and (iii) cloud merging leading to the formation of a bubble layer and, eventually, to the cone structure due to the radial pressure gradient on the horn tip.

Controlled tough bioadhesion mediated by ultrasound.

Tough bioadhesion has important implications in engineering and medicine but remains challenging to form and control. We report an ultrasound (US)-mediated strategy to achieve tough bioadhesion with controllability and fatigue resistance. Without chemical reaction, the US can amplify the adhesion energy and interfacial fatigue threshold between hydrogels and porcine skin by up to 100 and 10 times. Combined experiments and theoretical modeling suggest that the key mechanism is US-induced cavitation, which propels and immobilizes anchoring primers into tissues with mitigated barrier effects. Our strategy achieves spatial patterning of tough bioadhesion, on-demand detachment, and transdermal drug delivery. This work expands the material repertoire for tough bioadhesion and enables bioadhesive technologies with high-level controllability.

The effect of size range on ultrasound-induced translations in microbubble populations.

Microbubble translations driven by ultrasound-induced radiation forces can be beneficial for applications in ultrasound molecular imaging and drug delivery. Here, the effect of size range in microbubble populations on their translations is investigated experimentally and theoretically. The displacements within five distinct size-isolated microbubble populations are driven by a standard ultrasound-imaging probe at frequencies ranging from 3 to 7 MHz, and measured using the multi-gate spectral Doppler approach. Peak microbubble displacements, reaching up to 10 µm per pulse, are found to describe transient phenomena from the resonant proportion of each bubble population. The overall trend of the statistical behavior of the bubble displacements, quantified by the total number of identified displacements, reveals significant differences between the bubble populations as a function of the transmission frequency. A good agreement is found between the experiments and theory that includes a model parameter fit, which is further supported by separate measurements of individual microbubbles to characterize the viscoelasticity of their stabilizing lipid shell. These findings may help to tune the microbubble size distribution and ultrasound transmission parameters to optimize the radiation-force translations. They also demonstrate a simple technique to characterize the microbubble shell viscosity, the fitted model parameter, from freely floating microbubble populations using a standard ultrasound-imaging probe.

Microbubble Radiation Force-Induced Translation in Plane-Wave Versus Focused Transmission Modes.

Due to the primary radiation force, microbubble displacement has been observed previously in the focal region of single-element and array ultrasound probes. This effect has been harnessed to increase the contact between the microbubbles and targeted endothelium for drug delivery and ultrasound molecular imaging. In this study, microbubble displacements associated with plane-wave (PW) transmission are thoroughly investigated and compared to those obtained in focused-wave (FW) transmission over a range of pulse repetition frequencies, burst lengths (BLs), peak negative pressures, and transmission frequencies. In PW mode, the displacements, depending upon the experimental conditions, are in some cases consistently higher (e.g., by 28%, when the longest BL was used at PRF = 4 kHz), and the axial displacements are spatially more uniform compared to FW mode. Statistical analysis on the measured displacements reveals a slightly different frequency dependence of statistical quantities compared to transient peak microbubble displacements, which may suggest the need to consider the size range within the tested microbubble population.

Luminescence from cavitation bubbles deformed in uniform pressure gradients.

Presented here are observations that demonstrate how the deformation of millimetric cavitation bubbles by a uniform pressure gradient quenches single-collapse luminescence. Our innovative measurement system captures a broad luminescence spectrum (wavelength range, 300-900 nm) from the individual collapses of laser-induced bubbles in water. By varying the bubble size, driving pressure, and perceived gravity level aboard parabolic flights, we probed the limit from aspherical to highly spherical bubble collapses. Luminescence was detected for bubbles of maximum radii within the previously uncovered range, R_{0}=1.5-6 mm, for laser-induced bubbles. The relative luminescence energy was found to rapidly decrease as a function of the bubble asymmetry quantified by the anisotropy parameter ζ, which is the dimensionless equivalent of the Kelvin impulse. As established previously, ζ also dictates the characteristic parameters of bubble-driven microjets. The threshold of ζ beyond which no luminescence is observed in our experiment closely coincides with the threshold where the microjets visibly pierce the bubble and drive a vapor jet during the rebound. The individual fitted blackbody temperatures range between T_{lum}=7000 and T_{lum}=11500 K but do not show any clear trend as a function of ζ. Time-resolved measurements using a high-speed photodetector disclose multiple luminescence events at each bubble collapse. The averaged full width at half-maximum of the pulse is found to scale with R_{0} and to range between 10 and 20 ns.