Kuang Wei Lin

Effects of tissue stiffness, ultrasound frequency, and pressure on histotripsy-induced cavitation bubble behavior.

Histotripsy is an ultrasound ablation method that controls cavitation to fractionate soft tissue. In order to effectively fractionate tissue, histotripsy requires cavitation bubbles to rapidly expand from nanometer-sized initial nuclei into bubbles often larger than 50 µm. Using a negative pressure high enough to initiate a bubble cloud and expand bubbles to a sufficient size, histotripsy has been shown capable of completely fractionating soft tissue into acelluar debris resulting in effective tissue removal. Previous work has shown that the histotripsy process is affected by tissue mechanical properties with stiffer tissues showing increased resistance to histotripsy fractionation, which we hypothesize to be caused by impeded bubble expansion in stiffer tissues. In this study, the hypothesis that increases in tissue stiffness cause a reduction in bubble expansion was investigated both theoretically and experimentally. High speed optical imaging was used to capture a series of time delayed images of bubbles produced inside mechanically tunable agarose tissue phantoms using histotripsy pulses produced by 345 kHz, 500 kHz, 1.5 MHz, and 3 MHz histotripsy transducers. The results demonstrated a significant decrease in maximum bubble radius (Rmax) and collapse time (tc) with both increasing Youngs modulus and increasing frequency. Furthermore, results showed that Rmax was not increased by raising the pressure above the intrinsic threshold. Finally, this work demonstrated the potential of using a dual-frequency strategy to modulate the expansion of histotripsy bubbles. Overall, the results of this study improve our understanding of how tissue stiffness and ultrasound parameters affect histotripsy-induced bubble behavior and provide a rational basis to tailor acoustic parameters for treatment of the specific tissues of interest.

Effects of ultrasound frequency and tissue stiffness on the histotripsy intrinsic threshold for cavitation.

Histotripsy is an ultrasound ablation method that depends on the initiation of a cavitation bubble cloud to fractionate soft tissue. Previous work has indicated that a cavitation cloud can be formed by a single pulse with one high-amplitude negative cycle, when the negative pressure amplitude directly exceeds a pressure threshold intrinsic to the medium. We hypothesize that the intrinsic threshold in water-based tissues is determined by the properties of the water inside the tissue, and changes in tissue stiffness or ultrasound frequency will have a minimal impact on the histotripsy intrinsic threshold. To test this hypothesis, the histotripsy intrinsic threshold was investigated both experimentally and theoretically. The probability of cavitation was measured by subjecting tissue phantoms with adjustable mechanical properties and ex vivo tissues to a histotripsy pulse of 1-2 cycles produced by 345-kHz, 500-kHz, 1.5-MHz and 3-MHz histotripsy transducers. Cavitation was detected and characterized by passive cavitation detection and high-speed photography, from which the probability of cavitation was measured versus pressure amplitude. The results revealed that the intrinsic threshold (the negative pressure at which probability = 0.5) is independent of stiffness for Youngs moduli (E) <1 MPa, with only a small increase (∼2-3 MPa) in the intrinsic threshold for tendon (E = 380 MPa). Additionally, results for all samples revealed only a small increase of ∼2-3 MPa when the frequency was increased from 345 kHz to 3 MHz. The intrinsic threshold was measured to be between 24.7 and 30.6 MPa for all samples and frequencies tested in this study. Overall, the results of this study indicate that the intrinsic threshold to initiate a histotripsy bubble cloud is not significantly affected by tissue stiffness or ultrasound frequency in the hundreds of kilohertz to megahertz range.

Effects of Ultrasound Frequency on Nanodroplet-Mediated Histotripsy.

Nanodroplet-mediated histotripsy (NMH) is a targeted ultrasound ablation technique combining histotripsy with nanodroplets that can be selectively delivered to tumor cells for targeted tumor ablation. In a previous study, it was reported that by use of extremely short, high-pressure pulses, histotripsy cavitation bubbles were generated in regions containing nanodroplets at significantly lower pressure (∼10.8 MPa) than without nanodroplets (∼28 MPa) at 500 kHz. Furthermore, it was hypothesized that lower frequency would improve the effectiveness of NMH by increasing the size of the focal region, increasing bubble expansion, and decreasing the cavitation threshold. In this study, we investigated the effects of ultrasound frequency (345 kHz, 500 kHz, 1.5 MHz, and 3 MHz) on NMH. First, the NMH cavitation threshold was measured in tissue phantoms with and without nanodroplets, with results indicating that the NMH threshold was significantly below the histotripsy intrinsic threshold at all frequencies. Results also indicated that the NMH threshold decreased at lower frequency, ranging from 7.4 MPa at 345 kHz to 13.2 MPa at 3 MHz. In the second part of this study, the effects of frequency on NMH bubble expansion were investigated, with results indicating larger expansion at lower frequency, even at a lower pressure. In the final part of this study, the ability of perfluoropentane-encapsulated nanodroplets to act as sustainable cavitation nuclei over multiple pulses was investigated, with results indicating that the nanodroplets are destroyed by the cavitation process and only function as cavitation nuclei for the first few pulses, with this effect being most pronounced at higher frequencies. Overall, the results of this study support our hypothesis that using a lower frequency will improve the effectiveness of NMH by increasing the size of the focal region, increasing bubble expansion and decreasing the cavitation threshold.

Rapid prototyping fabrication of focused ultrasound transducers

Rapid prototyping (RP) fabrication techniques are currently widely used in diverse industrial and medical fields, providing substantial advantages in development time and costs in comparison to more traditional manufacturing processes. This paper presents a new method for the fabrication of high-intensity focused ultrasound transducers using RP technology. The construction of a large-aperture hemispherical transducer designed by computer software is described to demonstrate the process. The transducer was conceived as a modular design consisting of 32 individually focused 50.8-mm (2-in) PZT-8 element modules distributed in a 300-mm hemispherical scaffold with a geometric focus of 150 mm. The entire structure of the array, including the module housings and the hemispherical scaffold was fabricated through a stereolithography (SLA) system using a proprietary photopolymer. The PZT elements were bonded to the lenses through a quarter-wave tungsten-epoxy matching layer developed in-house specifically for this purpose. Modules constructed in this manner displayed a high degree of electroacoustic consistency, with an electrical impedance mean and standard deviation of 109 ± 10.2 Ω for the 32 elements. Time-of-flight measurements for individually pulsed modules mounted on the hemispherical scaffold showed that all pulses arrived at the focus within a 350 ns range, indicating a good degree of element alignment. Pressure profile measurements of the fully assembled transducer also showed close agreement with simulated results. The measured focal beam FWHM dimensions were 1.9 × 4.0 mm (1.9 × 3.9 mm simulated) in the transversal and axial directions respectively. Total material expenses associated with the construction of the transducer were approximately 5000 USD (as of 2011). The versatility and lower fabrication costs afforded by RP methods may be beneficial in the development of complex transducer geometries suitable for a variety of research and clinical applications.

Real-Time Feedback of Histotripsy Thrombolysis Using Bubble-Induced Color Doppler

Histotripsy thrombolysis is a non-invasive, drug-free, image-guided therapy that fractionates blood clots using well-controlled acoustic cavitation alone. Real-time quantitative feedback is highly desired during histotripsy thrombolysis treatment to monitor the progress of clot fractionation. Bubble-induced color Doppler (BCD) monitors the motion after cavitation generated by each histotripsy pulse, which has been found in gel and ex vivo liver tissue to be correlated with histotripsy fractionation. We investigated the potential of BCD to quantitatively monitor histotripsy thrombolysis in real time. To visualize clot fractionation, transparent three-layered fibrin clots were developed. Results indicated that a coherent motion follows the cavitation generated by each histotripsy pulse with a push and rebound pattern. The temporal profile of this motion expands and saturates as treatment progresses. A strong correlation exists between the degree of histotripsy clot fractionation and two metrics extracted from BCD: time of peak rebound velocity (tPRV) and focal mean velocity at a fixed delay (Vf,delay). The saturation of clot fractionation (i.e., treatment completion) matches well the saturations detected using tPRV and Vf,delay. The mean Pearson correlation coefficients between the progression of clot fractionation and the two BCD metrics were 93.1% and 92.6%, respectively. To validate BCD feedback in in vitro clots, debris volumes from histotripsy thrombolysis were obtained at different therapy doses and compared with Vf,delay. There is also good agreement between the increasing and saturation trends of debris volume and Vf,delay. Finally, a real-time BCD feedback algorithm to predict complete clot fractionation during histotripsy thrombolysis was developed and tested. This work illustrates the potential of BCD to monitor histotripsy thrombolysis treatment in real time.

Histotripsy Cardiac Therapy System Integrated with Real-Time Motion Correction

Histotripsy has shown promise in non-invasive cardiac therapy for neonatal and fetal applications. However, for cardiac applications in general, and especially in the adult heart, cardiac and respiratory motion may affect treatment accuracy and efficacy. In this article, we describe a histotripsy-mediated cardiac therapy system integrated with a fast motion tracking algorithm and treatment monitoring using ultrasound imaging. Motion tracking is performed by diamond search block matching in real-time ultrasound images using a reference image of the moving target, refined by Kalman filtering. As proof of feasibility, this algorithm was configured to track 2-D target motion and then electronically adjust the focus of a 1-MHz annular therapy array to correct for axial motion. This integrated motion tracking system is capable of sub-millimeter accuracy for displacements of 0-15 mm and velocities of 0-80 mm/s, with a maximum error less than 3 mm. Tissue phantom tests indicated that treatment efficiency and lesion size using motion tracking over displacements of 0-15 mm and velocities of 0-42 mm/s are comparable to those achieved when treating stationary targets. In vivo validation was conducted in an open-chest canine model, where the system provided 24 min of motion-corrected histotripsy therapy in the live beating heart, generating a targeted lesion on the atrial septum. Based on this proof of feasibility and the natural extension of these techniques to three dimensions, we anticipate a full motion correction system would be feasible and beneficial for non-invasive cardiac therapy.

Effects of Temperature on the Histotripsy Intrinsic Threshold for Cavitation

Histotripsy is an ultrasound ablation method that depends on the initiation of a dense cavitation bubble cloud to fractionate soft tissue. Previous work has demonstrated that a cavitation cloud can be formed by a single acoustic pulse with one high-amplitude negative cycle, when the negative pressure amplitude exceeds a threshold intrinsic to the medium. The intrinsic thresholds in soft tissues and tissue phantoms that are water based are similar to the intrinsic threshold of water over an experimentally verified frequency range of 0.3-3 MHz. Previous work studying the histotripsy intrinsic threshold has been limited to experiments performed at room temperature (~20 °C). In this study, we investigate the effects of temperature on the histotripsy intrinsic threshold in water, which is essential to accurately predict the intrinsic thresholds expected over the full range of in vivo therapeutic temperatures. Based on previous work studying the histotripsy intrinsic threshold and classical nucleation theory, we hypothesize that the intrinsic threshold will decrease with increasing temperature. To test this hypothesis, the intrinsic threshold in water was investigated both experimentally and theoretically. The probability of generating cavitation bubbles was measured by applying a single pulse with one high-amplitude negative cycle at 1 MHz to distilled degassed water at temperatures ranging from 10 °C to 90 °C. Cavitation was detected and characterized by passive cavitation detection and high-speed photography, from which the probability of cavitation was measured versus pressure amplitude. The results indicate that the intrinsic threshold (the negative pressure at which the cavitation probability = 0.5) significantly decreases with increasing temperature, showing a nearly linear decreasing trend from 29.8 ±0.4 MPa at 10 °C to 14.9 ± 1.4 MPa at 90 °C. Overall, the results of this study support our hypothesis that the intrinsic threshold is highly dependent on the temperature of the medium, which may allow for better predictions of cavitation generation at body temperature in vivo and at the elevated temperatures commonly seen in high-intensity focused ultrasound regimes.

Ultrasound backscatter spectral analysis provides image feedback for histotripsy tissue fractionation

The feasibility of using spectral analysis as an image feedback modality for histotripsy tissue fractionation was investigated in this study. Spectral analysis analyzes the frequency domain of the backscattered ultrasound RF signal, and its corresponding spectral parameters have been shown to have relationships with scatterer diameter and concentration. Ex vivo canine liver was dissected and embedded in 1.5% (w:v) agarose hydrogel, and then treated with histotripsy pulses driven at 2.0 MHz frequency with pulse repetition frequency of 100 Hz (PRF), pulse duration of 10 cycles, P+/P- = 31.8/17.4 MPa, and the number of pulses varied from 100 to 2000 pulses. After treatment, the specimens were scanned by a commercial ultrasound scanner, and the backscattered RF signals were collected. The power spectrum of the RF signals were analyzed by spectral analysis procedure with two approximation methods, one proposed by Lizzi et al. in 1996 [1], and the other by Oelze et al. in 2002 [2]. The results from both methods showed that the quantified scatterer diameter and acoustic concentration (=scatterer concentration × (relative acoustic impedance)2) decreased as the number of histotripsy treatment pulses increased. Furthermore, the quantified acoustic concentration had a strong correlation with the remaining nuclei density appeared in histological section of the treated specimen. These results suggest that the spectral analysis procedure could provide image feedback for histotripsy tissue fractionation.

A noninvasive approach for metastatic lymph node ablation using histotripsy tissue fractionation

Lymph node dissection is a widely used surgical procedure to treat and stage metastatic lymph nodes but has substantial side effects, such as lymphedema. Histotripsy is a noninvasive ultrasonic therapy that creates well-demarcated tissue fractionation using high-pressure and short acoustic shockwave pulses. This study investigates the possibility using of histotripsy to ablate lymph nodes noninvasively. Experiments were performed on 6 mixed breed healthy pigs, 11-12 weeks old and weighing 31-43 kg. Their superficial inguinal lymph nodes were treated in vivo with 5- or 10-cycle histotripsy pulses generated by focused, 1MHz therapy transducers, delivered at 50Hz pulse repetition frequency and peak rarefaction pressure above 21MPa. The treatment was guided by ultrasound imaging probes (either ATL CL15-7 or ATL L12-5) with a commercial ultrasound imaging system, HDI 5000. In one selected case, contrast-specific ultrasound was performed after perimammary injection of Sonazoid (GE Healthcare, Oslo, Norway) to localize the lymph nodes. Anatomical landmarks were then used to identify the same nodes for targeted therapy. The histological results show that, after histotripsy treatment, only the targeted lymph nodes were affected and they had demarcated foci of necrosis with minimal damage to the adjacent and intervening tissue. This in vivo acute study demonstrates the capability of histotripsy to noninvasively generate tissue fractionation within targeted lymph nodes, suggesting that histotripsy has the potential in noninvasive lymph node ablation.

Development of optical transmitters for high-amplitude focused ultrasound

This paper investigates the use of thin-film optical transmitters to generate focused ultrasound, aiming to develop highamplitude focused ultrasound. Composite films were used as the optoacoustic sources, which consist of carbonnanotubes (CNTs) and elastomeric polymers. As the nano-composites work as excellent optical absorbers and efficient heat converters, thermo-elastic volume deformation within the composites produces strong optoacoustic pressure. These films were formed on concave substrates for optoacoustic generation of the focused ultrasound. A focal waveform was measured using a single-mode fiber-optic hydrophone. A peak positive pressure of ~4 MPa was achieved.