Jin Xu

Experimental investigation of the effect of stiffness, exposure time and scan direction on the dimension of ultrasound histotripsy lesions.

Histotripsy uses high-intensity focused ultrasound to create energetic bubble clouds inside tissue to liquefy a region and has the advantages of higher contrast B-mode monitoring and sharp borders. This study experimentally investigated the effects of stiffness, exposure time and scan direction on the size of histotripsy-induced lesions in agar samples. A targeted region 0.45 cm wide (lateral) and 0.6 cm deep (axial) was scanned with the step sizes of 0.075 cm and 0.3 cm, respectively. The single-element spherically focused source (1.1 MHz, 6.34 cm focal length, f/1) had the peak compressional and rarefactional pressures of approximately 102 and 17 MPa. Pulses consisted of 20-cycle sine wave tone bursts with a burst period of 3 ms and exposure time of 15, 30 or 60 s. Also, both inward and outward scan direction were tested along the beam axis. The liquefied lesions generally had a larger size than the initially targeted region with larger sizes corresponding to softer agar and longer exposure. There was not a statistically significant difference in the lesion size with scan direction.

Minimization of treatment time for in vitro 1.1 MHz destruction of Pseudomonas aeruginosa biofilms by high-intensity focused ultrasound.

Medical implants are prone to colonization by bacterial biofilms. Normally, surgery is required to replace the infected implant. One promising noninvasive modality is to destroy biofilms with high-intensity focused ultrasound. In our study, Pseudomonas aeruginosa biofilms were grown on implant-mimicking graphite disks in a flow chamber for 3 days prior to exposing them to ultrasound pulses. Exposure time at each treatment location was varied between 5, 15 and 30s. Burst period was varied between 1, 3, 6 and 12 milliseconds (ms). The pulses were 20 cycles in duration at 1.1 MHz from a spherically focused transducer (f/1, 63 mm focal length), creating peak compressional and rarefactional pressures at the graphite disk surface of 30 and 13 MPa, respectively. P. aeruginosa were tagged with green fluorescent protein, and killed cells were visualized using propidium iodide before determining the extent of biofilm destruction. The exposure-induced temperature rise was measured to be less than 0.2°C at the focus, namely the interface between graphite disk and water. Then, the temperature rise was measured at the focus between the graphite disk and a tissue-mimicking phantom to evaluate therapy safety. Two thresholds, of bacteria destruction increase and of complete bacteria removal, respectively, were identified to divide our eight exposure conditions. Results indicated that 30-s exposure and 6-ms pulse period were sufficient to destroy the biofilms. However, the 15-s exposure and 3-ms pulse period were viewed as optimum when considering exposure time, efficacy, and safety.

Lysis of Chlamydomonas reinhardtii by high-intensity focused ultrasound as a function of exposure time

Efficient lysis of microalgae for lipid extraction is an important concern when processing biofuels. Historically, ultrasound frequencies in the range of 10-40 kHz have been utilized for this task. However, greater efficiencies might be achievable if higher frequencies could be used. In our study, we evaluated the potential of using 1.1 MHz ultrasound to lyse microalgae for biofuel production while using Chlamydomonas reinhardtii as a model organism. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at a pulse repetition frequency of 2 kHz (3.6% duty cycle). The time-average acoustic power output was 26.2 W while the spatial-peak-pulse-average intensity (ISPPA) for each tone burst was 41 kW/cm(2). The peak compressional and rarefactional pressures at the focus were 102 and 17 MPa, respectively. The exposure time was varied for the different cases in the experiments from 5s to 9 min and cell lysis was assessed by quantifying the percentage of protein and chlorophyll release into the supernate as well as the lipid extractability. Free radical generation and lipid oxidation for the different ultrasound exposures were also determined. We found that there was a statistically significant increase in lipid extractability for all of the exposures compared to the control. The longer exposures also completely fragmented the cells releasing almost all of the protein and chlorophyll into the supernate. The cavitation activity did not significantly increase lipid oxidation while there was a minor trend of increased free radical production with increased ultrasound exposure.

Impact of Preconditioning Pulse on Lesion Formation During High-Intensity Focused Ultrasound Histotripsy

Therapeutic applications with high-intensity focused ultrasound (HIFU) fall into two classifications-one using thermal effect for coagulation or ablation while generally avoiding cavitation and the other using cavitation-mediated mechanical effects while suppressing heating. Representative of the latter, histotripsy uses HIFU at low duty factor to create energetic bubble clouds inside tissue to liquefy a region and has the advantages in real-time monitoring and lesion fidelity to treatment planning. We explored the impact of a preconditioning/heating pulse on histotripsy lesion formation in porcine muscle samples. During sonication, a targeted square region 9 mm wide (lateral to the focal plane) was scanned in a raster pattern with a step size of 0.75 mm. The 20-s exposure at each treatment location consisted of a 5-s duration preconditioning burst at spatial-peak intensities from 0-1386 W/cm² followed by 5000 tone bursts at high intensity (with spatial-peak pulse-average intensity of 47.34 kW/cm², spatial-peak temporal-average intensity of 284 W/cm², peak compressional pressure of 102 MPa and peak rarefactional pressure of 17 MPa). The temperature increase for all exposures was measured using a thermal imager immediately after each exposure. Lesion volume increased with increasing amplitude of the preconditioning pulse until coagulation was observed, but lesion width/area did not change significantly with the amplitude. In addition, the lesion dimensions became smaller when the global tissue temperature was raised before applying the histotripsy pulsing sequence. Therefore, the benefit of the preconditioning pulse was not caused by global heating.

Dependence of ablative ability of high-intensity focused ultrasound cavitation-based histotripsy on mechanical properties of agar

Cavitation-based histotripsy uses high-intensity focused ultrasound at low duty factor to create bubble clouds inside tissue to liquefy a region, and provides better fidelity to planned lesion coordinates and the ability to perform real-time monitoring. The goal of this study was to identify the most important mechanical properties for predicting lesion dimensions, among these three: Youngs modulus, bending strength, and fracture toughness. Lesions were generated inside tissue-mimicking agar, and correlations were examined between the mechanical properties and the lesion dimensions, quantified by lesion volume and by the width and length of the equivalent bubble cluster. Histotripsy was applied to agar samples with varied properties. A cuboid of 4.5 mm width (lateral to focal plane) and 6 mm depth (along beam axis) was scanned in a raster pattern with respective step sizes of 0.75 and 3 mm. The exposure at each treatment location was either 15, 30, or 60 s. Results showed that only Youngs modulus influenced histotripsys ablative ability and was significantly correlated with lesion volume and bubble cluster dimensions. The other two properties had negligible effects on lesion formation. Also, exposure time differentially affected the width and depth of the bubble cluster volume.

Precision control of lesions by high-intensity focused ultrasound cavitation-based histotripsy through varying pulse duration

The goal of this experimental study was to explore the feasibility of acquiring controllable precision through varying pulse duration for lesions generated by cavitation-based histotripsy. Histotripsy uses high-intensity focused ultrasound (HIFU) at low duty factor to create energetic bubble clouds inside tissue to liquefy a region. It uses cavitation-mediated mechanical effects while minimizing heating, and has the advantages of real-time monitoring and lesion fidelity to treatment planning. In our study, histotripsy was applied to three groups of tissue-mimicking agar samples of different stiffnesses (29.4 ± 5.3, 44.8 ± 5.9, and 66.4 ± 7.1 kPa). B-mode imaging was used first to quantify bubble cluster dimensions in both water and agar. Then, a 4.5-mm-wide square (lateral to the focal plane) was scanned in a raster pattern with a step size of 0.75 mm in agar histotripsy experiments to estimate equivalent bubble cluster dimensions based on the histotripsyinduced damage. The 15-s exposure at each treatment location comprised 5000 sine-wave tone bursts at a spatial-peak pulseaverage intensity of 41.1 kW/cm2, with peak compressional and rarefactional pressures of 102 and 17 MPa, respectively. The results showed that bubble cluster width and length increased with pulse duration and decreased with agar stiffness. Therefore, a significant improvement in histotripsy precision could be achieved by reducing the pulse duration.

Effect of pulse repetition frequency and scan step size on the dimensions of the lesions formed in agar by HIFU histotripsy

Histotripsy uses high-intensity focused ultrasound pulses at low duty cycle to generate energetic bubble clouds inside tissue to fractionate a region. As a potential tumor treatment modality, this cavitation-based non-invasive technique has the advantages of easy monitoring and sharp borders. Aiming at therapy efficiency, we experimentally investigated the effects of pulse repetition frequency (PRF) and lateral scan step size on the dimensions of lesions formed through HIFU histotripsy in agar mimicking tissue in terms of mechanical (not acoustical) properties. The single-element spherically focused source (1.1 MHz, 6.34 cm focal length, f/1) was excited to reach the peak compressional and rarefactional pressures of ~102 and 17 MPa, respectively. A targeted rectangular block of 4.5 mm wide (lateral) and 6mm deep (axial) was scanned in a raster pattern with a constant axial step size of 3mm. The lateral step size was varied between 375, 750, 1500, 2250 and 4500 μm. Pulses at each treatment location consisted of 5000 20-cycle sine wave tone bursts with the PRF of 167, 333 or 1000 Hz. Results suggested that the bubble activity region could extend beyond the -3 dB region and that refining the lateral scan mesh and/or increasing PRF enlarged the lesion extent. The 1500 μm-333 Hz and the 1500 μm-1 kHz conditions were in a more favorable position to be viewed as optimal with regard to lesion volume generation rate, bubble activity region width, and the potential for thermal damage.

Assessment of Ultrasound Histotripsy–Induced Damage to Ex Vivo Porcine Muscle

Cavitation‐based histotripsy uses high‐intensity focused ultrasound pulses at a low duty cycle to generate energetic bubble clouds inside tissue to fractionate cells and is a potential noninvasive tumor treatment modality. Aiming at determining therapy efficiency, we experimentally investigated the effects of pulse repetition frequency and lateral scan step size on the degree of damage of histotripsy‐induced lesions in porcine muscle tissue.

Flow rate and duty cycle effects in lysis of Chlamydomonas reinhardtii using high-energy pulsed focused ultrasound

To consider microalgae lipid biofuel as a viable energy source, it is a necessity to maximize algal cell lysis, lipid harvest, and thus biofuel production versus the energy used to lyse the cells. Previous techniques have been to use energy consumptive ultrasound waves in the 10-40 kHz range in a stationary exposure environment. This study evaluated the potential of using 1.1 MHz ultrasound pulses in a new flow through type chamber on Chlamydomonas reinhardtii as a model organism for cell breakage. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at varied pulse repetition frequencies. First, variations in flow rate were examined at a constant duty cycle of 3.6%. After assessing flow rates, the duty cycle was varied to further explore the dependence on the tone burst parameters. Cell lysis was assessed by quantifying protein and chlorophyll release into the supernatant as well as by lipid extractability. Appropriate flow rates with higher duty cycles led to statistically significant increases in cell lysis relative to controls and other exposure conditions.

Mechanical destruction of pseudomonas aeruginosa biofilms by ultrasound exposure

Medical implants are prone to colonization by bacterial biofilms, which are highly resistant to antibiotics. Normally, surgery is required to replace the infected implant. One promising non-invasive treatment option is to destroy the biofilm with high-intensity focused ultrasound (HIFU) exposure. In our study, Pseudomonas aeruginosa bacterial biofilms were grown on graphite disks in a flow chamber for three days prior to exposing them to ultrasound pulses of varying duration or burst period. The pulses were 20 cycles in duration at a frequency of 1.1 MHz from a spherically focused transducer (f/1, 63 mm focal length), creating peak compressional and rarefactional pressures at the disk surface of 30 and 13 MPa, respectively. P. aeruginosa were tagged with GFP and cells killed by HIFU were visualized using propidium iodide, which permeates membranes of dead cells, to aid determining the extent of biofilm destruction and whether cells are alive or dead. Our results indicate that a 30-s exposure and 6-ms puls...