Microscopic residual bubble nuclei can persist on the order of 1 second following a cavitation event. These bubbles can limit the efficacy of ultrasound therapies such as shock wave lithotripsy and histotripsy, as they attenuate pulses that arrive subsequent to their formation and seed repetitive cavitation activity at a discrete set of sites (cavitation memory). Here, we explore a strategy for the removal of these residual bubbles following a cavitation event, using low amplitude ultrasound pulses to stimulate bubble coalescence. All experiments were conducted in degassed water and monitored using high speed photography. In each case, a 2 MHz histotripsy transducer was used to initiate cavitation activity (a cavitational bubble cloud), the collapse of which generated a population of residual bubble nuclei. This residual nuclei population was then sonicated using a 1 ms pulse from a separate 500 kHz transducer, which we term the ‘bubble removal pulse.’ Bubble removal pulse amplitudes ranging from 0 to 1.7 MPa were tested, and the backlit area of shadow from bubbles remaining in the field following bubble removal was calculated to quantify efficacy. It was found that an ideal amplitude range exists (roughly 180 – 570 kPa) in which bubble removal pulses stimulate the aggregation and subsequent coalescence of residual bubble nuclei, effectively removing them from the field. Further optimization of bubble removal pulse sequences stands to provide an adjunct to cavitation-based ultrasound therapies such as shock wave lithotripsy and histotripsy, mitigating the effects of residual bubble nuclei that currently limit their efficacy.

The efficacy of ultrasound therapies such as shock wave lithotripsy and histotripsy can be compromised by residual cavitation bubble nuclei that persist following the collapse of primary cavitation. In our previous work, we have developed a unique strategy for mitigating the effects of these residual bubbles using low amplitude ultrasound pulses to stimulate their aggregation and subsequent coalescence—effectively removing them from the field. Here, we further develop this bubble removal strategy through an investigation of the effect of frequency on the consolidation process. Bubble removal pulses ranging from 0.5 – 2 MHz were used to sonicate the population of residual nuclei produced upon collapse of a histotripsy bubble cloud. For each frequency, mechanical index (MI) values ranging from 0 to approximately 1.5 were tested. Results indicated that, when evaluated as a function of bubble removal pulse MI, the efficacy of bubble removal shows markedly similar trends for all frequencies tested. This behavior divides into three distinct regimes (with provided cutoffs being approximate): (1) MI < 0.2: Minimal effect on the population of remnant cavitation nuclei; (2) 0.2 < MI < 1: Aggregation and subsequent coalescence of residual bubbles, the extent of which trends toward a maximum; (3) MI > 1: Bubble coalescence is compromised as bubble removal pulses induce high magnitude inertial cavitation of residual bubbles. The major distinction in these trends came for bubble removal pulses applied at 2 MHz, which were observed to generate the most effective bubble coalescence of all frequencies tested. We hypothesize that this is a consequence of the secondary Bjerknes force being the major facilitator of the consolidation process, the magnitude of which increases when the bubble size distribution is far from resonance such that the phase difference of oscillation of individual bubbles is minimal.

Rate-dependent efficacy has been extensively documented in shock wave lithotripsy (SWL) stone comminution, with shock waves (SWs) delivered at a low rate producing more efficient fragmentation in comparison to those delivered at high rates. Cavitation is postulated to be the primary source underlying this rate phenomenon. Residual bubble nuclei that persist along the axis of SW propagation can drastically attenuate the waveform's negative phase, decreasing the energy which is ultimately delivered to the stone and compromising comminution. The effect is more pronounced at high rates, as residual nuclei have less time to passively dissolve between successive shocks. In this study, we investigate a means of actively removing such nuclei from the field using a low-amplitude acoustic pulse designed to stimulate their aggregation and subsequent coalescence. To test the efficacy of this bubble removal scheme, model kidney stones were treated in vitro using a research electrohydraulic lithotripter. SWL was applied at rates of 120, 60, or 30 SW/min with or without the incorporation of bubble removal pulses. Optical images displaying the extent of cavitation in the vicinity of the stone were also collected for each treatment. Results show that bubble removal pulses drastically enhance the efficacy of stone comminution at the higher rates tested (120 and 60 SW/min), while optical images show a corresponding reduction in bubble excitation along the SW axis when bubble removal pulses are incorporated. At the lower rate of 30 SW/min, no difference in stone comminution or bubble excitation was detected with the addition of bubble removal pulses, suggesting that remnant nuclei had sufficient time for more complete dissolution. These results corroborate previous work regarding the role of cavitation in rate-dependent SWL efficacy, and suggest that the effect can be mitigated via appropriate control of the cavitation environment surrounding the stone.

Residual bubbles produced after collapse of a cavitation cloud provide cavitation nuclei for subsequent cavitation events, causing cavitation to occur repeatedly at the same discrete set of sites. This effect, referred to as cavitation memory, limits the efficiency of histotripsy soft tissue fractionation. Besides passively mitigating cavitation memory by using a low pulse repetition frequency (~1 Hz), an active strategy was developed by our group. In this strategy, low-amplitude ultrasound sequences were used to stimulate coalescence of residual bubbles. The goal of this work is to remove cavitation memory and achieve rapid, homogeneous lesion formation using a single phased array transducer. A 1-MHz integrated histotripsy and bubble coalescing (BC) transducer system with a specialized electronic driving system was built in house. High-amplitude ( MPa) histotripsy pulses and subsequent low-amplitude (~1-2 MPa) BC sequences were applied to a red blood cell tissue-mimicking phantom at a single focal site. Significant reduction of the cavitation memory effect and increase in the fractionation rate were observed by introducing BC sequence. Effects of BC pulsing parameters were further studied. The optimal BC parameters were then utilized to homogenize a mm region at high rate.

Histotripsy is a tissue ablation method that utilizes focused, high-amplitude ultrasound to generate a cavitation bubble cloud that mechanically fractionates tissue. Effective histotripsy depends on the initiation, control, and maintenance of cavitation bubble clouds in the targeted area. In this study, we hypothesized that a low-pressure acoustic pulse sequence applied before and/or during histotripsy therapy would increase the cavitation initiation pressure threshold and the growth of cavitation bubble clouds. This technique could shrink or "sharpen" the focal zone during histotripsy to produce more precise and well-defined lesions with minimal collateral damage. It may also be a way to actively protect the soft tissue from cavitation damage during lithotripsy by increasing the pressure threshold for bubble cloud initiation. We applied these low-amplitude acoustic pulse sequences before and during histotripsy treatments with the pulse repetition frequency of 1 and 100 Hz, in three different mediums: water, tissue phantom agarose gel, and bovine liver in vitro. Acoustic backscatter signals and optical imaging were used to detect and monitor the initiation, maintenance, and growth of the resulting cavitation bubble cloud. The results demonstrated that the use of low-amplitude acoustic pulse sequences could increase the cavitation pressure amplitude threshold by 20% in the targeted area.