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Yak-Nam Wang

Senior Engineer

Email

ynwang@apl.washington.edu

Phone

206-616-6673

Videos

Mechanical Tissue Ablation with Focused Ultrasound

An experimental noninvasive surgery method uses nonlinear ultrasound pulses to liquefy tissue at remote target sites within a small focal region without damaging intervening tissues.

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23 Mar 2017

Boiling histotripsy utilizes sequences of millisecond-duration HIFU pulses with high-amplitude shocks that form at the focus by nonlinear propagation effects. Due to strong attenuation of the ultrasound energy at the shocks, these nonlinear waves rapidly heat tissue and generate millimeter-sized boiling bubbles at the focus within each pulse. Then the further interaction of subsequent shocks with the vapor cavity causes tissue disintegration into subcellular debris through the acoustic atomization mechanism.

The method was proposed at APL-UW in collaboration with Moscow State University (Russia) and now is being evaluated for various clinical applications. It has particular promise because of its important clinical advantages: the treatment of tissue volumes can be accelerated while sparing adjacent structures and not injuring intervening tissues; it generates precisely controlled mechanical lesions with sharp margins; the method can be implemented in existing clinical systems; and it can be used with real-time ultrasound imaging for targeting, guidance, and evaluation of outcomes. In addition, compared to thermal ablation, BH may lead to faster resorption of the liquefied lesion contents.

Non-invasive Treatment of Abscesses with Ultrasound

Abscesses are walled-off collections of fluid and bacteria within the body. They are common complications of surgery, trauma, and systemic infections. Typical treatment is the surgical placement of a drainage catheter to drain the abscess fluid over several days. Dr. Keith Chan and researchers at APL-UW's Center for Industrial + Medical Ultrasound are exploring how to treat abscesses non-invasively, that is, from outside the body, with high-intensity focused ultrasound (HIFU). This experimental therapy could reduce pain, radiation exposure, antibiotic use, and costs for patients with abscesses. Therapeutic ultrasound could also treat abscesses too small or inaccessible for conventional drainage.

20 Jun 2016

Publications

2000-present and while at APL-UW

Effect of carbon dioxide on the twinkling artifact in ultrasound imaging of kidney stones: A pilot study

Simon, J.C., Y.-N. Wang, B.W. Cunitz, J. Thiel, F. Starr, Z. Liu, and M.R. Bailey, "Effect of carbon dioxide on the twinkling artifact in ultrasound imaging of kidney stones: A pilot study," Ultrasound Med. Bill. 43, 877-883, doi:10.1016/j.ultrasmedbio.2016.12.010, 2017.

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1 May 2017

Bone demineralization, dehydration and stasis put astronauts at increased risk of forming kidney stones in space. The color-Doppler ultrasound "twinkling artifact," which highlights kidney stones with color, can make stones readily detectable with ultrasound; however, our previous results suggest twinkling is caused by microbubbles on the stone surface which could be affected by the elevated levels of carbon dioxide found on space vehicles. Four pigs were implanted with kidney stones and imaged with ultrasound while the anesthetic carrier gas oscillated between oxygen and air containing 0.8% carbon dioxide. On exposure of the pigs to 0.8% carbon dioxide, twinkling was significantly reduced after 9–25 min and recovered when the carrier gas returned to oxygen. These trends repeated when pigs were again exposed to 0.8% carbon dioxide followed by oxygen. The reduction of twinkling caused by exposure to elevated carbon dioxide may make kidney stone detection with twinkling difficult in current space vehicles.

Noninvasive characterization of pancreatic tumor mouse models using magnetic resonance imaging

Farr, N., Y.-N. Wang, S. D'Andrea, K.M. Gravelle, J.H. Hwang, and D. Lee, "Noninvasive characterization of pancreatic tumor mouse models using magnetic resonance imaging," Cancer Med., 6, 1082-1090, doi:10.1002/cam4.1062, 2017.

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1 May 2017

The preclinical models of pancreatic adenocarcinoma provide an alternative means for determining the mechanisms of malignancy and possibilities for treatments, thus representing a resource of immense potential for cancer treatment in medicine. To evaluate different tumor models, quantifiable magnetic resonance imaging (MRI) techniques can play a significant role in identifying valuable in vivo biomarkers of tumor characteristics. We characterized three models of pancreatic cancer with multiparametric MRI techniques. Tumor stromal density of each tumor was measured using diffusion-weighted imaging and magnetization transfer (MT-MRI). Histologic measurement showed a similar trend with tumor fibrosis levels. Results indicated that MRI measurements can serve as a valuable tool in identifying and evaluating tumor characteristics.

Release of cell-free microRNA biomarkers into the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy

Chevillet, J.R., and 9 others, including F. Starr and Y.-N. Wang, "Release of cell-free microRNA biomarkers into the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy," Radiology, 283, 158-167, doi:10.1148/radiol.2016160024, 2017.

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1 Apr 2017

Purpose
To compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics.

Materials and Methods
All rat experiments were approved by the University of Washington Institutional Animal Care and Use Committee. Reverse-transcription quantitative polymerase chain reaction array profiling was used to identify candidate microRNA biomarkers in a rat solid tumor cell line. Rats subcutaneously grafted with these cells were randomly assigned among three pulsed focused ultrasound treatment groups: (a) local tissue liquefaction via boiling histotripsy, (b) tissue permeabilization via inertial cavitation, and (c) mild (<10°C) heating of tissue, as well as a sham-treated control group. Blood specimens were drawn immediately prior to treatment and serially over 24 hours afterward. Plasma microRNA was quantified with reverse-transcription quantitative polymerase chain reaction, and statistical significance was determined with one-way analysis of variance (Kruskal-Wallis and Friedman tests), followed by the Dunn multiple-comparisons test.

Results
After tissue liquefaction and cavitation treatments (but not mild heating), plasma quantities of candidate biomarkers increased significantly (P value range, <.0001 to .04) relative to sham-treated controls. A threefold to 32-fold increase occurred within 15 minutes after initiation of pulsed focused ultrasound tumor treatment, and these increases persisted for 3 hours. Histologic examination confirmed complete liquefaction of the targeted tumor area with boiling histotripsy, in addition to areas of petechial hemorrhage and tissue disruption by means of cavitation-based treatment.

Conclusion
Mechanical tumor tissue disruption with pulsed focused ultrasound–induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment.

More Publications

Release of cell-free microRNA tumor biomarkers into the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy

Chevillet, J.R., T.D. Khokhlova, M.D. Giraldez, G.R. Shade, F. Starr, Y.-N. Wang, E.N. Gallichotte, K. Wang, J.H. Want, and M. Tewari, "Release of cell-free microRNA tumor biomarkers into the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy," Radiology, 283, 158-167, doi:10.1148/radiol.2016160024, 2017.

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1 Apr 2017

To compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics.

All rat experiments were approved by the University of Washington Institutional Animal Care and Use Committee. Reverse-transcription quantitative polymerase chain reaction array profiling was used to identify candidate microRNA biomarkers in a rat solid tumor cell line. Rats subcutaneously grafted with these cells were randomly assigned among three pulsed focused ultrasound treatment groups: (a) local tissue liquefaction via boiling histotripsy, (b) tissue permeabilization via inertial cavitation, and (c) mild (<10°C) heating of tissue, as well as a sham-treated control group. Blood specimens were drawn immediately prior to treatment and serially over 24 hours afterward. Plasma microRNA was quantified with reverse-transcription quantitative polymerase chain reaction, and statistical significance was determined with one-way analysis of variance (Kruskal-Wallis and Friedman tests), followed by the Dunn multiple-comparisons test.

After tissue liquefaction and cavitation treatments (but not mild heating), plasma quantities of candidate biomarkers increased significantly (P value range, <.0001 to .04) relative to sham-treated controls. A threefold to 32-fold increase occurred within 15 minutes after initiation of pulsed focused ultrasound tumor treatment, and these increases persisted for 3 hours. Histologic examination confirmed complete liquefaction of the targeted tumor area with boiling histotripsy, in addition to areas of petechial hemorrhage and tissue disruption by means of cavitation-based treatment.

Mechanical tumor tissue disruption with pulsed focused ultrasound–induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment.

Release of cell-free microRNA tumor biomarkers in the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy

Chevillet, J.R., T.D. Khokhlova, M.D. Giraldez, G.R. Schade, F. Starr, Y.-N. Wang, E.N. Gallichotte, K. Wang, J.H. Hwang, and M. Tewari, "Release of cell-free microRNA tumor biomarkers in the blood circulation with pulsed focused ultrasound: A noninvasive, anatomically localized, molecular liquid biopsy," Radiology, EOR, doi:10.1148/radiol.2016160024, 2016.

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1 Nov 2016

The purpose is to compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics. Mechanical tumor tissue disruption with pulsed focused ultrasound–induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment.

The association between mechanical and biochemical/histological characteristics in diabetic and non-diabetic plantar soft tissue

Ledoux, W.R., S. Pai, J.B. Shofer, and Y.-N. Wang, "The association between mechanical and biochemical/histological characteristics in diabetic and non-diabetic plantar soft tissue," J. Biomech., 49, 3328-3333, doi:10.1016/j.jbiomech.2016.08.021, 2016.

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3 Oct 2016

Diabetes, and the subsequent complication of lower limb ulcers leading to potential amputation, remains an important health care problem in United States, even with declining amputation rates. It has been well documented that diabetes can alter the mechanical properties (i.e., increased stiffness) of the plantar soft tissue, although this finding is not universal. Similarly, biochemical, and histological changes have been found in the plantar soft tissue, but, as with the mechanical changes, these findings are not consistent across all studies. Our group׳s work has demonstrated that diabetes increases plantar soft tissue modulus and increases elastic septal thickness. The purpose of the current study was to explore the association between mechanical, biochemical and histological properties. Using previously collected data, a linear mixed effects regression was conducted. The correlations were weak; of the 32 that were tested, only 3 (modulus to septal thickness when location was accounted for, energy loss to total collagen, and energy loss to collagen/elastin ratio) were statistically significant, none with an R2 greater than 0.10. The main differences in the means were increased tissue stiffness and increased septal wall thickness, both trends were supported in the literature. However, as the correlations were weak, it is likely that another unexamined biochemical factor (perhaps collagen crosslinking) is associated with the mechanical tissue changes.

Histotripsy liquefaction of large hematomas

Khokhlova, T.D., W.L. Monsky, Y.A. Haider, A.D. Maxwell, Y.-N. Wang, and T.J. Matula, "Histotripsy liquefaction of large hematomas," Ultrasound Med. Biol., 42, 1491-1498, doi:10.1016/j.ultrasmedbio.2016.01.020, 2016.

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1 Jul 2016

Intra- and extra-muscular hematomas result from repetitive injury as well as sharp and blunt limb trauma. The clinical consequences can be serious, including debilitating pain and functional deficit. There are currently no short-term treatment options for large hematomas, only lengthy conservative treatment. The goal of this work was to evaluate the feasibility of a high intensity focused ultrasound (HIFU)-based technique, termed histotripsy, for rapid (within a clinically relevant timeframe of 15–20 min) liquefaction of large volume (up to 20 mL) extra-vascular hematomas for subsequent fine-needle aspiration. Experiments were performed using in vitro extravascular hematoma phantoms—fresh bovine blood poured into 50 mL molds and allowed to clot. The resulting phantoms were treated by boiling histotripsy (BH), cavitation histotripsy (CH) or a combination in a degassed water tank under ultrasound guidance. Two different transducers operating at 1 MHz and 1.5 MHz with f-number = 1 were used. The liquefied lysate was aspirated and analyzed by histology and sized in a Coulter Counter. The peak instantaneous power to achieve BH was lower than (at 1.5 MHz) or equal to (at 1 MHz) that which was required to initiate CH. Under the same exposure duration, BH-induced cavities were one and a half to two times larger than the CH-induced cavities, but the CH-induced cavities were more regularly shaped, facilitating easier aspiration. The lysates contained a small amount of debris larger than 70 μm, and 99% of particulates were smaller than 10 μm. A combination treatment of BH (for initial debulking) and CH (for liquefaction of small residual fragments) yielded 20 mL of lysate within 17.5 minutes of treatment and was found to be most optimal for liquefaction of large extravascular hematomas.

An ultrasonic caliper device for measuring acoustic nonlinearity

Hunter, C., O.A Sapozhnikov, A.D. Maxwell, V.A. Khokhlova, Y.-N. Wang, B. MacConaghy, and W. Kreider, "An ultrasonic caliper device for measuring acoustic nonlinearity," Phys. Procedia, 87, 93-98, doi:10.1016/j.phpro.2016.12.015, 2016.

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1 May 2016

In medical and industrial ultrasound, it is often necessary to measure the acoustic properties of a material. A specific medical application requires measurements of sound speed, attenuation, and nonlinearity to characterize livers being evaluated for transplantation. For this application, a transmission-mode caliper device is proposed in which both transmit and receive transducers are directly coupled to a test sample, the propagation distance is measured with an indicator gage, and receive waveforms are recorded for analysis. In this configuration, accurate measurements of nonlinearity present particular challenges: diffraction effects can be considerable while nonlinear distortions over short distances typically remain small. To enable simple estimates of the nonlinearity coeffcient from a quasi-linear approximation to the lossless Burgers’ equation, the calipers utilize a large transmitter and plane waves are measured at distances of 15–50 mm. Waves at 667 kHz and pressures between 0.1 and 1 MPa were generated and measured in water at different distances; the nonlinearity coeffcient of water was estimated from these measurements with a variability of approximately 10%. Ongoing efforts seek to test caliper performance in other media and improve accuracy via additional transducer calibrations.

Enhancement of small molecule delivery by pulsed high-intensity focused ultrasound: A parameter exploration

Zhou, Y., Y.-N. Wang, N. Farr, J. Zia, H. Chen, B.M. Ko, T. Khokhlova, T. Li, and J.H. Hwang, "Enhancement of small molecule delivery by pulsed high-intensity focused ultrasound: A parameter exploration," Ultrasound Med. Biol., 42, 956-963, doi:10.1016/j.ultrasmedbio.2015.12.009, 2016.

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1 Apr 2016

Chemotherapeutic drug delivery is often ineffective within solid tumors, but increasing the drug dose would result in systemic toxicity. The use of high-intensity focused ultrasound (HIFU) has the potential to enhance penetration of small molecules. However, operation parameters need to be optimized before the use of chemotherapeutic drugs in vivo and translation to clinical trials. In this study, the effects of pulsed HIFU (pHIFU) parameters (spatial-average pulse-average intensity, duty factor and pulse repetition frequency) on the penetration as well as content of small molecules were evaluated in ex vivo porcine kidneys. Specific HIFU parameters resulted in more than 40 times greater Evans blue content and 3.5 times the penetration depth compared with untreated samples. When selected parameters were applied to porcine kidneys in vivo, a 2.3-fold increase in concentration was obtained after a 2-min exposure to pHIFU. Pulsed HIFU has been found to be an effective modality to enhance both the concentration and penetration depth of small molecules in tissue using the optimized HIFU parameters. Although, performed in normal tissue, this study has the promise of translation into tumor tissue.

Pulsed high-intensity focused ultrasound enhances delivery of doxorubicin in a preclinical model of pancreatic cancer

Li, T. Y.-N. Wang, T.D. Khokhlova, S. D'Andrea, F. Starr, H. Chen, J.S. McCune, L.J. Risler, A. Mashadi-Hossein, and J.H. Hwang, "Pulsed high-intensity focused ultrasound enhances delivery of doxorubicin in a preclinical model of pancreatic cancer," Cancer Res., 75, 3738-3746, doi:10.1158/0008-5472.CAN-15-0296, 2015.

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15 Sep 2015

Pancreatic cancer is characterized by extensive stromal desmoplasia, which decreases blood perfusion and impedes chemotherapy delivery. Breaking the stromal barrier could both increase perfusion and permeabilize the tumor, enhancing chemotherapy penetration. Mechanical disruption of the stroma can be achieved using ultrasound-induced bubble activity-cavitation. Cavitation is also known to result in microstreaming and could have the added benefit of actively enhancing diffusion into the tumors. Here, we report the ability to enhance chemotherapeutic drug doxorubicin penetration using ultrasound-induced cavitation in a genetically engineered mouse model (KPC mouse) of pancreatic ductal adenocarcinoma. To induce localized inertial cavitation in pancreatic tumors, pulsed high-intensity focused ultrasound (pHIFU) was used either during or before doxorubicin administration to elucidate the mechanisms of enhanced drug delivery (active vs. passive drug diffusion). For both types, the pHIFU exposures that were associated with high cavitation activity resulted in disruption of the highly fibrotic stromal matrix and enhanced the normalized doxorubicin concentration by up to 4.5-fold compared with controls. Furthermore, normalized doxorubicin concentration was associated with the cavitation metrics (P < 0.01), indicating that high and sustained cavitation results in increased chemotherapy penetration. No significant difference between the outcomes of the two types, that is, doxorubicin infusion during or after pHIFU treatment, was observed, suggesting that passive diffusion into previously permeabilized tissue is the major mechanism for the increase in drug concentration. Together, the data indicate that pHIFU treatment of pancreatic tumors when resulting in high and sustained cavitation can efficiently enhance chemotherapy delivery to pancreatic tumors.

Endoscopic high-intensity focused US: Technical aspects and studies in an in vivo porcine model

Li, T., T. Khokhlova, E. Maloney, Y.-N. Wang, S. D'Andrea, F. Starr, N. Farr, K. Morrison, G. Keilman, and J.H. Hwang, "Endoscopic high-intensity focused US: Technical aspects and studies in an in vivo porcine model," Gastrointest. Endoscopy, 81, 1243-1250, doi:0.1016/j.gie.2014.12.019, 2015.

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1 May 2015

High-intensity focused US (HIFU) is becoming more widely used for noninvasive and minimally invasive ablation of benign and malignant tumors. Recent studies suggest that HIFU can also enhance targeted drug delivery and stimulate an antitumor immune response in many tumors. However, targeting pancreatic and liver tumors by using an extracorporeal source is challenging due to the lack of an adequate acoustic window. The development of an EUS-guided HIFU transducer has many potential benefits including improved targeting, decreased energy requirements, and decreased potential for injury to intervening structures.

The transducer successfully created lesions in gel phantoms and ex vivo bovine livers. In vivo studies demonstrated that targeting and creating lesions in the porcine pancreas and liver are feasible. An EUS-guided HIFU transducer was successfully designed and developed with dimensions that are appropriate for endoscopic use. The feasibility of performing EUS-guided HIFU ablation in vivo was demonstrated in an in vivo porcine model. Further development of this technology will allow endoscopists to perform precise therapeutic ablation of periluminal lesions without breaching the wall of the gastric tract.

Investigation into the mechanisms of tissue atomization by high-intensity focused ultrasound

Simon, J.C., O.A. Sapzhnikov, Y.-N. Wang, V.A. Khokhlova, L.A. Crum, and M.R. Bailey, "Investigation into the mechanisms of tissue atomization by high-intensity focused ultrasound," Ultrasound Med. Biol., 41, 1372-1385, doi:10.1016/j.ultrasmedbio.2014.12.022, 2015.

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1 May 2015

Ultrasonic atomization, or the emission of a fog of droplets, was recently proposed to explain tissue fractionation in boiling histotripsy. However, even though liquid atomization has been studied extensively, the mechanisms underlying tissue atomization remain unclear. In the work described here, high-speed photography and overpressure were used to evaluate the role of bubbles in tissue atomization. As static pressure increased, the degree of fractionation decreased, and the ex vivo tissue became thermally denatured. The effect of surface wetness on atomization was also evaluated in vivo and in tissue-mimicking gels, where surface wetness was found to enhance atomization by forming surface instabilities that augment cavitation. In addition, experimental results indicated that wetting collagenous tissues, such as the liver capsule, allowed atomization to breach such barriers. These results highlight the importance of bubbles and surface instabilities in atomization and could be used to enhance boiling histotripsy for transition to clinical use.

Histotripsy methods in mechanical disintegration of tissue: Toward clinical applications

Khokhlova, V.A., J.B. Fowlkes, W.W. Roberts, G.R. Schade, Z. Xu, T.D. Khokhlova, T.L. Hall, A.D. Maxwell, Y.-N. Wang, and C.A. Cain, "Histotripsy methods in mechanical disintegration of tissue: Toward clinical applications," Int. J. Hypertherm., 31, 145-162, doi:10.3109/02656736.2015.1007538, 2015.

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1 Mar 2015

In high intensity focused ultrasound (HIFU) therapy, an ultrasound beam is focused within the body to locally affect the targeted site without damaging intervening tissues. The most common HIFU regime is thermal ablation. Recently there has been increasing interest in generating purely mechanical lesions in tissue (histotripsy). This paper provides an overview of several studies on the development of histotripsy methods toward clinical applications. Two histotripsy approaches and examples of their applications are presented. In one approach, sequences of high-amplitude, short (microsecond-long), focused ultrasound pulses periodically produce dense, energetic bubble clouds that mechanically disintegrate tissue. In an alternative approach, longer (millisecond-long) pulses with shock fronts generate boiling bubbles and the interaction of shock fronts with the resulting vapour cavity causes tissue disintegration. Recent preclinical studies on histotripsy are reviewed for treating benign prostatic hyperplasia (BPH), liver and kidney tumours, kidney stone fragmentation, enhancing anti-tumour immune response, and tissue decellularisation for regenerative medicine applications. Potential clinical advantages of the histotripsy methods are discussed. Histotripsy methods can be used to mechanically ablate a wide variety of tissues, whilst selectivity sparing structures such as large vessels. Both ultrasound and MR imaging can be used for targeting and monitoring the treatment in real time. Although the two approaches utilise different mechanisms for tissue disintegration, both have many of the same advantages and offer a promising alternative method of non-invasive surgery.

Preclinical safety and effectiveness studies of ultrasonic propulsion of kidney stones

Harper, J.D., B. Dunmire, Y.-N. Wang, J.C. Simon, D. Liggitt, M. Paun, B.W. Cunitz, F. Starr, M.R. Bailey, K.L. Penniston, F.C. Lee, R.S. Hsi, and M.D. Sorensen, "Preclinical safety and effectiveness studies of ultrasonic propulsion of kidney stones," Urology, 84, 484-489, doi:10.1016/j.urology.2014.04.041, 2014.

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1 Aug 2014

Objective
To provide an update on a research device to ultrasonically reposition kidney stones transcutaneously. This article reports preclinical safety and effectiveness studies, survival data, modifications of the system, and testing in a stone-forming porcine model. These data formed the basis for regulatory approval to test the device in humans.

Materials and Methods
The ultrasound burst was shortened to 50 ms from previous investigations with 1-s bursts. Focused ultrasound was used to expel 2- to 5-mm calcium oxalate monohydrate stones placed ureteroscopically in 5 pigs. Additionally, de novo stones were imaged and repositioned in a stone-forming porcine model. Acute safety studies were performed targeting 2 kidneys (6 sites) and 3 pancreases (8 sites). Survival studies followed 10 animals for 1 week after simulated treatment. Serum and urine analyses were performed, and tissues were evaluated histologically.

Results
All ureteroscopically implanted stones (6/6) were repositioned out of the kidney in 14 ± 8 minutes with 13 ± 6 bursts. On average, 3 bursts moved a stone more than 4 mm and collectively accounted for the majority of relocation. Stones (3 mm) were detected and repositioned in the 200-kg stone-forming model. No injury was detected in the acute or survival studies.

Conclusion
Ultrasonic propulsion is safe and effective in the porcine model. Stones were expelled from the kidney. De novo stones formed in a large porcine model were repositioned. No adverse effects were identified with the acute studies directly targeting kidney or pancreatic tissue or during the survival studies indicating no evidence of delayed tissue injury.

Passive cavitation detection during pulsed HIFU exposures of ex vivo tissues and in vivo mouse pancreatic tumors

Li, T., H. Chen, T. Khokhlova, Y.-N. Wang, W. Kreider, X. He, and J.H. Hwang, "Passive cavitation detection during pulsed HIFU exposures of ex vivo tissues and in vivo mouse pancreatic tumors," Ultrasound Med. Biol., 40, 1523-1543, doi:10.1016/j.ultrasmedbio.2014.01.007, 2014.

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1 Jul 2014

Pulsed high-intensity focused ultrasound (pHIFU) has been shown to enhance vascular permeability, disrupt tumor barriers and enhance drug penetration into tumor tissue through acoustic cavitation. Monitoring of cavitation activity during pHIFU treatments and knowing the ultrasound pressure levels sufficient to reliably induce cavitation in a given tissue are therefore very important. Here, three metrics of cavitation activity induced by pHIFU and evaluated by confocal passive cavitation detection were introduced: cavitation probability, cavitation persistence and the level of the broadband acoustic emissions.

These metrics were used to characterize cavitation activity in several ex vivo tissue types (bovine tongue and liver and porcine adipose tissue and kidney) and gel phantoms (polyacrylamide and agarose) at varying peak-rare factional focal pressures (1–12 MPa) during the following pHIFU protocol: frequency 1.1 MHz, pulse duration 1 ms and pulse repetition frequency 1 Hz. To evaluate the relevance of the measurements in ex vivo tissue, cavitation metrics were also investigated and compared in the ex vivo and in vivo murine pancreatic tumors that develop spontaneously in transgenic KrasLSL.G12 D/+; p53 R172 H/+ ; PdxCretg/ (KPC) mice and closely re-capitulate human disease in their morphology.

The cavitation threshold, defined at 50% cavitation probability, was found to vary broadly among the investigated tissues (within 2.5–10 MPa), depending mostly on the water-lipid ratio that characterizes the tissue composition. Cavitation persistence and the intensity of broadband emissions depended both on tissue structure and lipid concentration. Both the cavitation threshold and broadband noise emission level were similar between ex vivo and in vivo pancreatic tumor tissue. The largest difference between in vivo and ex vivo settings was found in the pattern of cavitation occurrence throughout pHIFU exposure: it was sporadic in vivo, but it decreased rapidly and stopped over the first few pulses ex vivo. Cavitation activity depended on the interplay between the destruction and circulation of cavitation nuclei, which are not only used up by HIFU treatment but also replenished or carried away by circulation in vivo. These findings are important for treatment planning and optimization in pHIFU-induced drug delivery, in particular for pancreatic tumors.

Ultrasound-guided tissue fractionation by high intensity focused ultrasound in an in vivo porcine liver model

Khokhlova, T.D., Y.-N. Wang, J.C. Simon, B.W. Cunitz, F. Starr, M. Paun, L.A. Crum, M.R. Bailey, and V.A. Khokhlova, "Ultrasound-guided tissue fractionation by high intensity focused ultrasound in an in vivo porcine liver model," P. Natl. Acad. Sci. USA, 111, 8161-8166, doi:10.1073/pnas.1318355111, 2014.

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3 Jun 2014

The clinical use of high intensity focused ultrasound (HIFU) therapy for noninvasive tissue ablation has been recently gaining momentum. In HIFU, ultrasound energy from an extracorporeal source is focused within the body to ablate tissue at the focus while leaving the surrounding organs and tissues unaffected. Most HIFU therapies are designed to use heating effects resulting from the absorption of ultrasound by tissue to create a thermally coagulated treatment volume. Although this approach is often successful, it has its limitations, such as the heat sink effect caused by the presence of a large blood vessel near the treatment area or heating of the ribs in the transcostal applications. HIFU-induced bubbles provide an alternative means to destroy the target tissue by mechanical disruption or, at its extreme, local fractionation of tissue within the focal region. Here, we demonstrate the feasibility of a recently developed approach to HIFU-induced ultrasound-guided tissue fractionation in an in vivo pig model. In this approach, termed boiling histotripsy, a millimeter-sized boiling bubble is generated by ultrasound and further interacts with the ultrasound field to fractionate porcine liver tissue into subcellular debris without inducing further thermal effects. Tissue selectivity, demonstrated by boiling histotripsy, allows for the treatment of tissue immediately adjacent to major blood vessels and other connective tissue structures. Furthermore, boiling histotripsy would benefit the clinical applications, in which it is important to accelerate resorption or passage of the ablated tissue volume, diminish pressure on the surrounding organs that causes discomfort, or insert openings between tissues.

Pulsed focused ultrasound treatment of muscle mitigates paralysis-induced bone loss in the adjacent bone: A study in a mouse model

Poliachik, S.L., T.D. Khokhlova, Y.-N. Wang, J.C. Simon, and M.R. Bailey, "Pulsed focused ultrasound treatment of muscle mitigates paralysis-induced bone loss in the adjacent bone: A study in a mouse model," Ultrasound Med. Biol., 40, 2113-2124, doi:10.1016/j.ultrasmedbio.2014.02.027, 2014.

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21 May 2014

Bone loss can result from bed rest, space flight, spinal cord injury or age-related hormonal changes. Current bone loss mitigation techniques include pharmaceutical interventions, exercise, pulsed ultrasound targeted to bone and whole body vibration. In this study, we attempted to mitigate paralysis-induced bone loss by applying focused ultrasound to the midbelly of a paralyzed muscle. We employed a mouse model of disuse that uses onabotulinumtoxinA-induced paralysis, which causes rapid bone loss in 5 d. A focused 2 MHz transducer applied pulsed exposures with pulse repetition frequency mimicking that of motor neuron firing during walking (80 Hz), standing (20 Hz), or the standard pulsed ultrasound frequency used in fracture healing (1 kHz). Exposures were applied daily to calf muscle for 4 consecutive d. Trabecular bone changes were characterized using micro-computed tomography. Our results indicated that application of certain focused pulsed ultrasound parameters was able to mitigate some of the paralysis-induced bone loss.

Focused ultrasound to displace renal calculi: Threshold for tissue injury

Wang, Y.-N., J.C. Simon, B.W. Cunitz, F.L. Starr, M. Paun, D.H. Liggitt, A.P. Evan, J.A. McAteer, Z. Liu, B. Dunmire, and M.R. Bailey, "Focused ultrasound to displace renal calculi: Threshold for tissue injury," J. Therapeut. Ultrasound, 2, doi:10.1186/2050-5736-2-5, 2014.

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31 Mar 2014

The global prevalence and incidence of renal calculi is reported to be increasing. Of the patients that undergo surgical intervention, nearly half experience symptomatic complications associated with stone fragments that are not passed and require follow-up surgical intervention. In a clinical simulation using a clinical prototype, ultrasonic propulsion was proven effective at repositioning kidney stones in pigs. The use of ultrasound to reposition smaller stones or stone fragments to a location that facilitates spontaneous clearance could therefore improve stone-free rates. The goal of this study was to determine an injury threshold under which stones could be safely repositioned.

Kidneys of 28 domestic swine were treated with exposures that ranged in duty cycle from 0%–100% and spatial peak pulse average intensities up to 30 kW/cm2 for a total duration of 10 min. The kidneys were processed for morphological analysis and evaluated for injury by experts blinded to the exposure conditions.

At a duty cycle of 3.3%, a spatial peak intensity threshold of 16,620 W/cm2 was needed before a statistically significant portion of the samples showed injury. This is nearly seven times the 2,400-W/cm2 maximum output of the clinical prototype used to move the stones effectively in pigs.

The data obtained from this study show that exposure of kidneys to ultrasonic propulsion for displacing renal calculi is well below the threshold for tissue injury.

Comparison of tissue injury from focused ultrasonic propulsion of kidney stones versus extracorporeal shock wave lithotripsy

Connors, B.A., A.P. Evan, P.M. Blomgren, R.S. Hsi, J.D. Harper, M.D. Sorensen, Y.-N. Wang, J.C. Simon, M. Paun, F. Starr, B.W. Cunitz, M.R. Bailey, and J.E. Lingeman, "Comparison of tissue injury from focused ultrasonic propulsion of kidney stones versus extracorporeal shock wave lithotripsy," J. Urol., 191, 235-241, doi:10.1016/j.juro.2013.07.087, 2014.

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1 Jan 2014

Focused ultrasonic propulsion is a new noninvasive technique designed to move kidney stones and stone fragments out of the urinary collecting system. However, to our knowledge the extent of tissue injury associated with this technique is not known. We quantitated the amount of tissue injury produced by focused ultrasonic propulsion under simulated clinical treatment conditions and under conditions of higher power or continuous duty cycles. We compared those results to extracorporeal shock wave lithotripsy injury.

Ultrasound intensity to propel stones from the kidney is below the threshold for renal injury

Wang, Y.-N., J.C. Simon, B. Cunitz, F. Starr, M. Paun, D. Liggit, A. Evan, J. McAteer, J. Williams, Z. Liu, P. Kaczkowski, R. Hsi, M. Sorensen, J. Harper, and M.R. Bailey, "Ultrasound intensity to propel stones from the kidney is below the threshold for renal injury," Proc., Meetings on Acoustics, 19, 075066, doi:10.1121/1.4800361, 2013.

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3 Jun 2013

Therapeutic ultrasound has an increasing number of applications in urology, including shockwave lithotripsy, stone propulsion, tissue ablation, and hemostasis. However, the threshold of renal injury using ultrasound is unknown. The goal of this study was to determine kidney injury thresholds for a range of intensities between diagnostic and ablative therapeutic ultrasound. A 2 MHz annular array generating spatial peak pulse average intensities (ISPPA) up to 28,000 W/cm2 in water was placed on the surface of in vivo porcine kidneys and focused on the adjacent parenchyma. Treatments consisted of pulses of 100 μs duration triggered every 3 ms for 10 minutes at various intensities. The perfusion-fixed tissue was scored by 3 blinded independent experts. Above a threshold of 16,620 W/cm2, the majority of injury observed included emulsification, necrosis and hemorrhage. Below this threshold, almost all injury presented as focal cell and tubular swelling and/or degeneration. These findings provide evidence for a wide range of potentially therapeutic ultrasound intensities that has a low probability of causing injury. While this study did not examine all combinations of treatment parameters of therapeutic ultrasound, tissue injury appears dose-dependent.

Focused ultrasound to expel calculi from the kidney: Safety and efficacy of a clinical prototype device

Harper, J.D., M.D. Sorensen, B.W. Cunitz, Y.-N. Wang, J.C. Simon, F. Starr, M. Paun, B. Dunmire, H.D. Liggitt, A.P. Evan, J.A. McAteer, R.S. Hsi, and M.R. Bailey, "Focused ultrasound to expel calculi from the kidney: Safety and efficacy of a clinical prototype device," J. Urol., 190, 1090-1095, doi:10.1016/j.juro.2013.03.120, 2013.

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9 Apr 2013

Purpose
Focused ultrasound has the potential to expel small stones or residual stone fragments from the kidney, or move obstructing stones to a non-obstructing location. The purpose of this study was to evaluate the efficacy and safety of ultrasonic propulsion in a live porcine model.

Material and Methods
Calcium oxalate monohydrate kidney stones and laboratory model stones (2–8 mm) were ureteroscopically implanted within the renal pelvicalyceal system of 12 kidneys in eight domestic swine. Transcutaneous ultrasonic propulsion was performed using a Philips HDI C5-2 imaging transducer and Verasonics diagnostic ultrasound platform. Successful stone relocation was defined as stone movement from the calyx to the renal pelvis, ureteropelvic junction (UPJ) or proximal ureter. Efficacy and procedure time were determined. Three blinded experts evaluated for histologic injury to the kidney in control, sham, and treatment arms.

Results
All stones were observed to move during treatment, and 65% (17/26) were relocated successfully to the renal pelvis (3), UPJ (2), or ureter (12). Average successful procedure time was 14±8 min and required 23±16 ultrasound bursts of ~1 sec duration. There was no evidence of gross or histologic injury to the renal parenchyma in kidneys exposed to 20 bursts (1 sec duration, 33 sec intervals) at the same output (2400 W/cm2) used to push stones.

Conclusions
Non-invasive transcutaneous ultrasonic propulsion is a safe, effective, and time-efficient means to relocate calyceal stones to the renal pelvis, UPJ, or ureter. This technology holds promise as a useful adjunct to the surgical management of renal calculi.

Histological and biochemical analysis of mechanical and thermal bioeffects in boiling histotripsy lesions induced by high intensity focused ultrasound

Wang, Y.-N., T. Khokhlova, M. Bailey, J.H. Hwang, and V. Khokhlova, "Histological and biochemical analysis of mechanical and thermal bioeffects in boiling histotripsy lesions induced by high intensity focused ultrasound," Ultrasound Med. Biol., 39, 424-438, doi:10.1016/j.ultrasmedbio.2012.10.012, 2013.

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1 Mar 2013

Recent studies have shown that shockwave heating and millisecond boiling in high-intensity focused ultrasound fields can result in mechanical fractionation or emulsification of tissue, termed boiling histotripsy. Visual observations of the change in color and contents indicated that the degree of thermal damage in the emulsified lesions can be controlled by varying the parameters of the exposure. The goal of this work was to examine thermal and mechanical effects in boiling histotripsy lesions using histologic and biochemical analysis. The lesions were induced in ex vivo bovine heart and liver using a 2-MHz single-element transducer operating at duty factors of 0.005–0.01, pulse durations of 5–500 ms and in situ shock amplitude of 73 MPa. Mechanical and thermal damage to tissue was evaluated histologically using conventional staining techniques (hematoxylin and eosin, and nicotinamide adenine dinucleotide-diaphorase). Thermal effects were quantified by measuring denaturation of salt soluble proteins in the treated region. According to histologic analysis, the lesions that visually appeared as a liquid contained no cellular structures larger than a cell nucleus and had a sharp border of one to two cells. Both histologic and protein analysis showed that lesions obtained with short pulses (<10 ms) did not contain any thermal damage. Increasing the pulse duration resulted in an increase in thermal damage. However, both protein analysis and nicotinamide adenine dinucleotide-diaphorase staining showed less denaturation than visually observed as whitening of tissue. The number of high-intensity focused ultrasound pulses delivered per exposure did not change the lesion shape or the degree of thermal denaturation, whereas the size of the lesion showed a saturating behavior suggesting optimal exposure duration. This study confirmed that boiling histotripsy offers an effective, predictable way to non-invasively fractionate tissue into sub-cellular fragments with or without inducing thermal damage.

Synthesis and characterization of anti-EGFR fluorescent nanoparticles for optical molecular imaging

Chan, L.W., Y.-N. Wang, L.Y. Lin, M.P. Upton, J.H. Hwang, and S.H. Pun, "Synthesis and characterization of anti-EGFR fluorescent nanoparticles for optical molecular imaging," Bioconjugate Chem., 24, 167-175, doi:10.1021/bc300355y, 2013.

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20 Feb 2013

Molecular imaging, the visualization of molecular and cellular markers, is a promising method for detection of dysplasia and early cancer in the esophagus and can potentially be used to identify regions of interest for biopsy or tumor margins for resection. EGFR is a previously reported cell surface receptor with stepwise increases in expression during the progression from Barrett's metaplasia to adenocarcinoma. In this work, a 200 nm fluorescent nanoparticle contrast agent was synthesized for targeted imaging of EGFR through a series of surface modifications to dye-encapsulated polystyrene particles. Amino-functionalized polystyrene particles were PEGylated using a heterobifunctional PEG linker. Subsequently, thiolated M225 antibodies were conjugated to maleimide functional groups on attached PEGs for EGFR targeting. In vitro binding studies using flow cytometry demonstrated specific binding of M225-PEG-NP to EGFR-expressing cells with minimal nonspecific binding in EGFR cells. Binding was shown to increase proportionally with the number of conjugated M225 antibodies. Adsorbed formulations with unmodified M225 antibodies, M225 PEG-NP, were synthesized using the same antibody feeds used in M225-PEG-NP synthesis to determine the contribution of adsorbed antibodies to EGFR targeting. Adsorbed antibodies were less efficient at mediated nanoparticle targeting to EGFR than conjugated antibodies. Finally, M225-PEG-NP demonstrated binding to EGFR-expressing regions in human esophageal tissue sections.

Ultrasonic atomization of tissue and its role in tissue fractionation by high intensity focused ultrasound

Simon, J.C., O.A. Sapozhnikov, V.A. Khokhlova, Y.-N. Wang, L.A. Crum, and M.R. Bailey, "Ultrasonic atomization of tissue and its role in tissue fractionation by high intensity focused ultrasound," Phys. Med. Biol. 57, 8061-8078, doi:10.1088/0031-9155/57/23/8061, 2012.

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7 Dec 2012

Atomization and fountain formation is a well-known phenomenon that occurs when a focused ultrasound wave in liquid encounters an air interface. High intensity focused ultrasound (HIFU) has been shown to fractionate a tissue into submicron-sized fragments in a process termed boiling histotripsy, wherein the focused ultrasound wave superheats the tissue at the focus, producing a millimetre-sized boiling or vapour bubble in several milliseconds. Yet the question of how this millimetre-sized boiling bubble creates submicron-sized tissue fragments remains. The hypothesis of this work is that the tissue can behave as a liquid such that it atomizes and forms a fountain within the vapour bubble produced in boiling histotripsy. We describe an experiment, in which a 2 MHz HIFU transducer (maximum in situ intensity of 24,000 W cm-2) was aligned with an air–tissue interface meant to simulate the boiling bubble. Atomization and fountain formation was observed with high-speed photography and resulted in tissue erosion. Histological examination of the atomized tissue showed whole and fragmented cells and nuclei. Air–liquid interfaces were also filmed. Our conclusion was that HIFU can fountain and atomize tissue. Although this process does not entirely mimic what was observed in liquids, it does explain many aspects of tissue fractionation in boiling histotripsy.

Novel high-intensity focused ultrasound clamp — potential adjunct for laparoscopic partial nephrectomy

Harper, J.D., A. Shah, S.B. Mitchell, Y.N. Wang, F. Starr, M.R. Bailey, and L.A. Crum, "Novel high-intensity focused ultrasound clamp — potential adjunct for laparoscopic partial nephrectomy," J. Endourol., 26, 1494-1499, doi:10.1098/end.2012.0107, 2012.

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1 Nov 2012

BACKGROUND AND PURPOSE:
Partial nephrectomy (PN) can be technically challenging, especially if performed in a minimally invasive manner. Although ultrasound technology has been shown to have therapeutic capabilities, including tissue ablation and hemostasis, it has not gained clinical use in the PN setting. The purpose of this study is to evaluate the ability of a high-intensity ultrasound clamp to create an ablation plane in the kidney providing hemostasis that could potentially aid in laparoscopic PN.
METHODS:
A new instrument was created using a laparoscopic Padron endoscopic exposing retractor. Ultrasound elements were engineered on both sides of the retractor to administer high-intensity ultrasound energy between the two sides of the clamp. This high-intensity focused ultrasound (HIFU) clamp was placed 2 to 2.5 cm from the upper and lower poles of 10 porcine kidneys to evaluate its effectiveness at different levels and duration of energy delivery. PN transection was performed through the distal portion of the clamped margin. Kidneys postintervention and after PN were evaluated and blood loss estimated by weighing gauze placed at the defect. Histologic analysis was performed with hematoxylin and eosin and nicotinamide adenine dinucleotide staining to evaluate for tissue viability and thermal spread.
RESULTS:
Gross parenchymal changes were seen with obvious demarcation between treated and untreated tissue. Increased ultrasound exposure time (10 vs 5 and 2 min), even at lower power settings, was more effective in causing destruction and necrosis of tissue. Transmural ablation was achieved in three of four renal units after 10 minutes of exposure with significantly less blood loss (<2 g vs 30-100 g). Nonviable tissue was confirmed histologically. There was minimal thermal spread outside the clamped margin (1.2-3.2 mm).
CONCLUSION:
In this preliminary porcine evaluation, a novel HIFU clamp induced hemostasis and created an ablation plane in the kidney. This technology could serve as a useful adjunct to laparoscopic PN in the future and potentially obviate the need for renal hilar clamping.

Modulation of gene expression using electrospun scaffolds with templated architecture

Karchin, A., Y.-N. Wang, and J.E. Sanders, "Modulation of gene expression using electrospun scaffolds with templated architecture," J. Biomed. Mater. Res. Part A, 100A, 1605-1614, doi:10.1002/jbm.a.34102, 2012.

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23 Mar 2012

The fabrication of biomimetic scaffolds is a critical component to fulfill the promise of functional tissue-engineered materials. We describe herein a simple technique, based on printed circuit board manufacturing, to produce novel templates for electrospinning scaffolds for tissue-engineering applications. This technique facilitates fabrication of electrospun scaffolds with templated architecture, which we defined as a scaffold's bulk mechanical properties being driven by its fiber architecture. Electrospun scaffolds with templated architectures were characterized with regard to fiber alignment and mechanical properties. Fast Fourier transform analysis revealed a high degree of fiber alignment along the conducting traces of the templates. Mechanical testing showed that scaffolds demonstrated tunable mechanical properties as a function of templated architecture. Fibroblast-seeded scaffolds were subjected to a peak strain of 3 or 10% at 0.5 Hz for 1 h. Exposing seeded scaffolds to the low strain magnitude (3%) significantly increased collagen I gene expression compared to the high strain magnitude (10%) in a scaffold architecture-dependent manner. These experiments indicate that scaffolds with templated architectures can be produced, and modulation of gene expression is possible with templated architectures. This technology holds promise for the long-term goal of creating tissue-engineered replacements with the biomechanical and biochemical make-up of native tissues.

Focused ultrasound to expel calculi from the kidney

Shah, A., J.D. Harper, B.W. Cunitz, Y.-N. Wang, M. Paun, J.C. Simon, W. Lu, P.J. Kaczkowski, and M.R. Bailey, "Focused ultrasound to expel calculi from the kidney," J. Urol., 187, 739-743, doi:10.1016/j.juro.2011.09.144, 2012.

1 Feb 2012

A method of mechanical emulsification in a bulk tissue using shock wave heating and millisecond boiling

Khokhlova, V.A., M.S. Canney, M.R. Bailey, J.H. Hwang, T.D. Khokhlova, W. Kreider, Y.N. Wang, J.C. Simon, Y. Zhou, O.A. Sapozhnikov, and L.A. Crum, "A method of mechanical emulsification in a bulk tissue using shock wave heating and millisecond boiling," J. Acoust. Soc. Am., 129, 2476, doi:10.1121/1.3588143, 2011.

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1 Apr 2011

Recent studies in high intensity focused ultrasound (HIFU) have shown significant interest in generating purely mechanical damage of tissue without thermal coagulation. Here, an approach using millisecond bursts of ultrasound shock waves and repeated localized boiling is presented. In HIFU fields, nonlinear propagation effects lead to formation of shocks only in a small focal region. Significant enhancement of heating due to absorption at the shocks leads to boiling temperatures in tissue in milliseconds as calculated based on weak shock theory. The heated and potentially necrotized region of tissue is small compared to the volume occupied by the mm-sized boiling bubble it creates. If the HIFU pulse is only slightly longer than the time-to-boil, thermal injury is negligible compared to the mechanical injury caused by the exploding boiling bubble and its further interaction with shocks. Experiments performed in transparent gels and various ex vivo and in vivo tissues have confirmed the effectiveness of this emulsification method. In addition, since mm-sized boiling bubbles are highly echogenic, tissue emulsification can be easily monitored in real-time using B-mode ultrasound imaging.

Effect of low intensity pulsed ultrasound on mesenchymal stem cells

Ruan, J.L., Y.N. Wang, L.A. Crum, and S.B. Mitchell, "Effect of low intensity pulsed ultrasound on mesenchymal stem cells," J. Acoust. Soc. Am., 129, 2576, doi:10.1121/1.3588505, 2011.

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1 Apr 2011

Low intensity pulsed ultrasound (LIPUS) has been used to accelerate tissue regeneration; however, the biological mechanisms of LIPUS induced regeneration is not completely understood. The aim for this study is to elucidate the mechanical effect generated by US for the stimulation of mesenchymal stem cells (MSCs). MSCs were cultured on flexible cell culture membranes and stimulated by US for 10 min daily with acoustic intensities of 0, 6, 13.5, and 22.5 W/cm2. Cell proliferation and viability were evaluated by direct cell count and Alamar Blue assay. Morphological evaluation was performed and cell-matrix interactions were evaluated. Cell-matrix interaction was analyzed by immunochemical staining of focal adhesion proteins. LIPUS enhanced cell proliferation at higher intensities and there was an increase in cell viability after 4 consecutive days of US treatment. No morphological changes were observed in all treatments. Expression of focal adhesion protein, vinculin, was enhanced after 3 consecutive days of ultrasound treatment. Studies of media agitation did not show any enhancement effect in cell proliferation or focal adhesion protein expression. The results validates that US is able to influence the cell matrix interaction. Application of higher acoustic pressure on cell growth environment can stimulate MSC proliferation and focal adhesion.

Histological and biochemical analysis of emulsified lesions in tissue induced by high intensity focused ultrasound

Wang, Y.N., T.D. Khokhlova, M.S. Canney, V.A. Khokhlova, L.A. Crum, and M.R. Bailey, "Histological and biochemical analysis of emulsified lesions in tissue induced by high intensity focused ultrasound," J. Acoust. Soc. Am., 129, 2477, doi:10.1121/1.3588148, 2011.

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1 Apr 2011

As recently shown, shock wave heating and millisecond boiling can be used to obtain mechanical emulsification of tissue with or without evident thermal damage, which can be controlled by varying the parameters of the high intensity focused ultrasound exposure. The goal of this work was to examine these bioeffects using histological and biochemical analysis. Lesions were created in ex vivo bovine heart and liver using a 2-MHz transducer and pulsing scheme with 71 MPa in situ shock amplitude, 0.01 duty factor, and 5-500 ms pulse duration. Mechanical tissue damage and viability of cells in the lesions were evaluated histologically using conventional staining techniques (H&E and NADH-diaphorase). Thermal effects were quantified by measuring denaturation of salt soluble proteins in the treated area and confirmed by histology. By visual observation, the liquefied lesions obtained with shorter pulses (< 15 ms) did not show any thermal damage that correlated well with the results of both histology and protein analysis. Increasing the pulse duration resulted in an increase in thermal damage; both protein analysis and NADH-diaphorase staining showed denaturation that was visually observed as whitening of the lesion content.

In vivo tissue emulsification using millisecond boiling induced by high intensity focused ultrasound

Khokhlova, T.D., J.C. Simon, Y.-N. Wang, V.A. Khokhlova, M. Paun, F.L. Starr, P.J. Kaczkowski, L.A. Crum, J.H. Hwang, and M.R. Bailey, "In vivo tissue emulsification using millisecond boiling induced by high intensity focused ultrasound," J. Acoust. Soc. Am., 129, 2477, doi:10.1121/1.3588149, 2011.

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1 Apr 2011

Shock-wave heating and millisecond boiling in high intensity focused ultrasound fields have been shown to result in mechanical emulsification of ex-vivo tissue. In this work, the same in situ exposures were applied in vivo in pig liver and in mice bearing 5-7 mm subcutaneous tumors (B16 melanoma) on the hind limb. Lesions were produced using a 2-MHz annular array in the case of pig liver (shock amplitudes up to 98 MPa) and a 3.4-MHz single-element transducer in the case of mouse tumors (shock amplitude of 67 MPa). The parameters of the pulsing protocol (1-500 ms pulse durations and 0.01-0.1 duty factor) were varied depending on the extent of desired thermal effect. All exposures were monitored using B-mode ultrasound. Mechanical and thermal tissue damage in the lesions was evaluated histologically using H&E and NADH-diphorase staining. The size and shape of emulsified lesions obtained in-vivo agreed well with those obtained in ex-vivo tissue samples using the same exposure parameters. The lesions were successfully produced both in bulk liver tissue at depths of 1-2 cm and in superficial tumors at depths less than 1 mm without damaging the skin.

Prototype for expulsion of kidney stones with focused ultrasound

Shah, A., J.D. Harper, B.W. Cunitz, J.C. Kucewicz, Y.N. Wang, J.C. Simon, W. Lu, P.J. Kaczkowski, and M.R. Bailey, "Prototype for expulsion of kidney stones with focused ultrasound," J. Acoust. Soc. Am., 129, 2376, doi:10.1121/1.3587694, 2011.

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1 Apr 2011

Residual fragments remain in over 50% of treatments for lower pole kidney stones. A second-generation device based on a diagnostic ultrasound system and scanhead has been developed with a unique algorithm for stone detection and the capability to focus ultrasound to expel residual fragments. Focused ultrasound was applied to a bead on string in a water tank as well as to human stones (<5 mm) implanted in the lower pole of a live porcine model via retrograde ureteroscopy. Histological samples were collected and scored in a blinded fashion for therapeutic exposures and for super-therapeutic levels. The in-vitro bead was visually observed to move under focused ultrasound. Even with progressive manual displacement of the bead, the system continuously tracked and caused bead movement in real time. In the live porcine model, stones were expelled from the lower pole to the ureteropelvic junction in seconds to minutes using pulses at a duty factor of 0.02 and 8 W total acoustic power. Injury was observed no more frequently than in controls. Occurrence of injury rose slightly above control at a duty factor of 0.02 and 80 W and at a duty factor of 1 and 8 W.

High intensity ultrasound activation of coagulation factors

Mitchell, S.B., N.C. Juaire, and Y.-N. Wang, "High intensity ultrasound activation of coagulation factors," J. Acoust. Soc. Am., 123, 3222, 2008.

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1 May 2008

Although it has been demonstrated that High Intensity Focused Ultrasound (HIFU) can induce vascular cauterization, acoustic hemostasis technology has not been successful in making the transition from proof-of-concept to clinical settings. One reason for this lack of acceptance is the limited understanding of the fundamental mechanisms involved in ultrasound–vessel and ultrasound–blood interactions. The aim of this research was to investigate the hematological and biochemical mechanisms which are influenced by HIFU induced coagulation. HIFU was used to induce coagulation in an in vitro hematological flow system and in animal models. The flow circuit and in vivo arteries were instrumented with flow probes and thermocouples in the treatment region. Physiological parameters were recorded throughout the in vivo experiments. Blood samples were drawn for analysis prior to, during, and immediately following each HIFU treatment. Clotting time, complete blood count, and biochemical analysis were performed immediately and citrated samples were immediately centrifuged and frozen for future analysis. Results indicate that HIFU can change local and systemic levels of thombin/anti-thrombin complex (TAT) and tissue plasminogen activator (tPA), as well as induce changes in activated clotting time (ACT). These results indicate that HIFU can induce coagulation via the coagulation cascades (TAT) and that normal hematological response to thrombus formation is unaffected.

Therapeutic ultrasound induced cell death from a histological perspective

Brayman, A., P. Kaczkowski, Y.-N. Wang, M. Andrew, L.A. Crum, S. Kargl, and G. Speyer, "Therapeutic ultrasound induced cell death from a histological perspective," J. Acoust. Soc. Am., 123, 2996, doi:10.1121/1.2932547, 2008.

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1 May 2008

High-power, short-exposure time, High Intensity Focused Ultrasound (HIFU) treatment protocols are under development that offer the potential to increase procedure throughput and optimize individual therapies. Histological examination and optical image analysis of tissues following dynamic HIFU exposure in ex vivo bovine liver have revealed that cells undergo a fundamentally different form of cell death. The rapid temperature rise due to the HIFU exposure leaves the cells structurally intact but no longer viable, similar to the cell "fixation" induced by snap-freezing. These results suggest that careful choice of both staining technique and metric for determining cell death are important in quantifying type and morphology of cell ablation, and more broadly, safety and efficacy of treatment. This finding is similar to those obtained and under discussion in the laser and RF ablation communities. Specifically, the NADH staining technique is superior to H&E for assessing cell viability, and an alternative measure of cell death may be preferable to the binary thermal dose threshold currently the standard for HIFU treatment.

Inventions

MRI-Guided Lithotripsy of Urinary Tract Stones

Record of Invention Number: 47984

Mike Bailey, Wayne Kreider, Adam Maxwell, Yak-Nam Wang

Disclosure

23 Feb 2017

Methods of soft tissue emulsification using a mechanism of ultrasonic atomization inside gas or vapor cavities and associated systems and devices

Patent Number: 9,498,651

Oleg Sapozhnikov, Mike Bailey, Larry Crum, Vera Khokhlova, Yak-Nam Wang

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Patent

22 Nov 2016

The present technology is directed to methods of soft tissue emulsification using a mechanism of ultrasonic atomization inside gas or vapor cavities, and associated systems and devices. In several embodiments, for example, a method of non-invasively treating tissue includes pulsing ultrasound energy from the ultrasound source toward the target site in tissue. The ultrasound source is configured to emit high intensity focused ultrasound (HIFU) waves. The target site comprises a pressure-release interface of a gas or vapor cavity located within the tissue. The method continues by generating shock waves in the tissue to induce a lesion in the tissue at the target site. The method additionally includes characterizing the lesion based on a degree of at least one of a mechanical or thermal ablation of the tissue.

Noninvasive Disintegration of Peyronie's Plaque with Focused Ultrasound

Record of Invention Number: 47233

Hunter Wessells, Mike Bailey, Mahri Haider, Tatiana Khokhlova, Frank Lee, Adam Maxwell, George Schade, Yak-Nam Wang

Disclosure

23 Feb 2015

More Inventions

Methods to Selectively Fragment and Remove Tissue While Sparing Extracellular Matrix, Vessels and Similar Structures Using Microsecond-long High Intensity Focused Ultrasound Pulses with Shocks

Record of Invention Number: 46742

Yak-Nam Wang, Mike Bailey, Vera Khokhlova, Tanya Khokhlova, Wayne Kreider, Adam Maxwell

Disclosure

18 Nov 2013

Method of Trabecular Bone Loss Mitigation Through Exposure of Adjacent Muscle to Pulsed Ultrasound of Moderate Intensity

Record of Invention Number: 46674

Mike Bailey, Tanya Khokhlova, Julianna Simon, Yak-Nam Wang

Disclosure

26 Sep 2013

Acoustic Disruption and Deactivation of Biofilms in Catheters

Record of Invention Number: 45929

Yak-Nam Wang, Mike Bailey, Francesco Curra

Disclosure

15 Jan 2012

Enhanced Drug Delivery Using Endoscopic HIFU

Record of Invention Number: 8507D

Yak-Nam Wang, Joo Ha Hwang, Yufeng Zhou

Disclosure

29 Oct 2009

Acoustics Air-Sea Interaction & Remote Sensing Center for Environmental & Information Systems Center for Industrial & Medical Ultrasound Electronic & Photonic Systems Ocean Engineering Ocean Physics Polar Science Center
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