Liposomes-based Nanoplatform Enlarges Ultrasound-related Diagnostic and Therapeutic Precision.

Ultrasound (US) is recognized in the medical field as a safe and effective imaging modality due to its lack of ionizing radiation, non-invasive approach, and real-time monitoring capability. Accompanying recent progress in nanomedicine, US has been providing hope of theranostic capability not only for imaging-based diagnosis but also for US-based therapy by taking advantage of the bioeffects induced by US. Cavitation, sonoporation, thermal effects, and other cascade effects stimulated by acoustic energy conversion have contributed to medical problem-solving in the past decades, although to varying degrees of efficacy in comparison to other methods. Recently, the usage of liposomesbased nanoplatform fuels the development of nanomedicine and provides novel clinical strategies for antitumor, thrombolysis, and controlled drug release. The merging of novel liposome-based nanoplatforms and US-induced reactions has promise for a new blueprint for future medicine. In the present review article, the value of liposome-based nanoplatforms in US-related diagnosis and therapy will be discussed and summarized along with potential future directions for further investigations.

Gas Embolization in a Rodent Model of Hepatocellular Carcinoma Using Acoustic Droplet Vaporization.

Trans-arterial embolization is a commonly used therapy in unresectable hepatocellular carcinoma. Current methods involve the careful placement of an intraarterial catheter and the deposition of embolizing particles. Gas embolotherapy has been proposed as an embolization method with the potential for high spatial resolution without the need for a catheter. This method involves vaporizing intravenouslyadministered droplets into gas bubbles using focused ultrasound - a process termed acoustic droplet vaporization. The bubbles can become lodged in the vasculature, thereby creating an embolus. Here, we initially demonstrate the feasibility of achieving significant targeted embolization with this method in the rat cremaster using intravital microscopy. The therapy was then tested in an ectopic xenograft mouse model of hepatocellular carcinoma. Gas embolotherapy was shown to maintain the tumor volume at baseline over a twoweek treatment course while control groups showed significant tumor growth. These preliminary results demonstrate thatgas embolotherapy could serve as an effective noninvasive method for the management of unresectable hepatocellular carcinoma.

Quantitative Ultrasound Imaging to Assess the Biceps Brachii Muscle in Chronic Post-Stroke Spasticity: Preliminary Observation.

We prospectively investigated the feasibility of using quantitative ultrasound imaging (QUI) to assess the biceps brachii muscle (BBM) in individuals with chronic post-stroke spasticity. To quantify muscle echogenicity and stiffness, we measured QUI parameters (gray-scale pixel value and shear wave velocity [SWV, m/s]) of the BBM in three groups: 16 healthy BBMs; 12 post-stroke, non-spastic BBMs; and 12 post-stroke, spastic BBMs. The QUI results were compared with the Modified Ashworth Scale and Tardieu Scale. A total of 20 SWVs were measured in each BBM, once at elbow in 90° flexion and again at maximally achievable extension using acoustic radiation force impulse imaging. BBM pixel value was measured in gray-scale images captured at 90° elbow flexion using ImageJ software. Statistical analyses included analysis of variance for examining the difference in SWV and pixel values among the three groups; Bonferroni correction for testing the difference in SWV and pixel values in a paired group; t-test for examining the difference in SWV values measured at two elbow angles; and Pearson correlation coefficient for analyzing the correlation of QUI to Modified Ashworth Scale and Tardieu Scale. SWV significantly differed between spastic BBMs and non-spastic or healthy BBMs. For pixel values, each of the three groups significantly differed from the others at elbow 90° flexion. The difference in SWV measured between the two elbow angles was also significant (p <0.01). A strong negative correlation was found between SWV and passive range of motion (R2 = -0.88, p <0.0001) in spastic upper limbs. These results suggest that the use of QUI is feasible in quantitative assessment of spastic BBM.

Effects of Droplet Composition on Nanodroplet-Mediated Histotripsy.

Nanodroplet-mediated histotripsy (NMH) is a targeted ablation technique combining histotripsy with nanodroplets that can be selectively delivered to tumor cells. In two previous studies, polymer-encapsulated perfluoropentane nanodroplets were used to generate well-defined ablation similar to that obtained with histotripsy, but at significantly lower pressure, when NMH therapy was applied at a pulse repetition frequency (PRF) of 10 Hz. However, cavitation was not maintained over multiple pulses when ultrasound was applied at a lower PRF (i.e., 1-5 Hz). We hypothesized that nanodroplets with a higher-boiling-point perfluorocarbon core would provide sustainable cavitation nuclei, allowing cavitation to be maintained over multiple pulses, even at low PRF, which is needed for efficient and complete tissue fractionation via histotripsy. To test this hypothesis, we investigated the effects of droplet composition on NMH therapy by applying histotripsy at various frequencies (345 kHz, 500 kHz, 1.5 MHz, 3 MHz) to tissue phantoms containing perfluoropentane (PFP, boiling point ∼29°C, surface tension ∼9.5 mN/m) and perfluorohexane (PFH, boiling point ∼56°C, surface tension ∼11.9 mN/m) nanodroplets. First, the effects of droplet composition on the NMH cavitation threshold were investigated, with results revealing a significant decrease (>10 MPa) in the peak negative pressure (p-) cavitation threshold for both types of nanodroplets compared with controls. A slight decrease (∼1-3 MPa) in threshold was observed for PFP phantoms compared with PFH phantoms. Next, the ability of nanodroplets to function as sustainable cavitation nuclei over multiple pulses was investigated, with results revealing that PFH nanodroplets were sustainable cavitation nuclei over 1,000 pulses, whereas PFP nanodroplets were destroyed during the first few pulses (<50 pulses), likely because of the lower boiling point. Finally, tissue phantoms containing a layer of embedded red blood cells were used to compare the damage generated for NMH treatments using PFP and PFH droplets, with results indicating that PFH nanodroplets significantly improved NMH ablation, allowing for well-defined lesions to be generated at all frequencies and PRFs tested. Overall, the results of this study provide significant insight into the role of droplet composition in NMH therapy and provide a rational basis to tailor droplet parameters to improve NMH tissue fractionation.

The role of positive and negative pressure on cavitation nucleation in nanodroplet-mediated histotripsy.

Nanodroplet-mediated histotripsy (NMH) is an ultrasound ablation technique combining histotripsy with acoustically sensitive perfluorocarbon (PFC) nanodroplets that can be selectively delivered to tumor cells for targeted tumor ablation. NMH takes advantage of the significantly reduced cavitation threshold of the nanodroplets, allowing for cavitation to be selectively generated only in regions containing nanodroplets. Understanding the physical mechanisms underlying the nanodroplet cavitation process is essential to the development of NMH. In this study, we hypothesize that cavitation nucleation is caused by the negative pressure (p-) exposed to the PFC, and the NMH cavitation threshold is therefore determined by the incident p- of the single-cycle pulses commonly used in NMH. This paper reports the first study that separately investigates the effects of negative and positive pressure on the NMH cavitation threshold using near half-cycle ultrasound pulses with dominant negative (negative-polarity pulses) or positive (positive-polarity pulses) pressure phases. Tissue phantoms containing perfluorohexane (PFH) nanodroplets were exposed to negative-polarity and positive-polarity pulses generated by a frequency compounding transducer recently developed in our lab, and the probability of generating cavitation was measured as a function of peak negative (p-) and peak positive (p+) pressure. The results showed close agreement in the p- cavitation threshold for PFH phantoms exposed to negative-polarity (11.4 ± 0.1 MPa) and positive-polarity (11.7 ± 0.2 MPa) pulses. The p+ at the cavitation threshold, in contrast, was measured to be sign ficantly different for the negative-polarity (4.0 ± 0.1 MPa) and positive-polarity (42.6 ± 0.2 MPa) pulses. In the final part of this study, the experimental results were compared to the cavitation threshold predicted by classical nucleation theory (CNT), with results showing close agreement between simulations and experiments. Overall, the results support our hypothesis and provide significant insight into the physical mechanisms underlying NMH.

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.

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 Young's 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.

[Perfluoropentane droplets enhanced HIFU therapeutic effect].

A small HIFU system was used to investigate the phase-shift droplet vaporization in vivo and its effect on thermal absorption in tissue-mimicking phantoms. The experiments demonstrated that droplets could be vaporized to bubbles in vivo by the small HIFU system and the volume of bubbles could increase by tens of times. With appropriate droplets concentration, lesion volume produced by HIFU could be increased significantly under the same HIFU parameter.

Ultrafast dynamics of the acoustic vaporization of phase-change microdroplets.

Acoustically sensitive emulsions are a promising tool for medical applications such as localized drug delivery. The physical mechanisms underlying the ultrasound-triggered nucleation and subsequent vaporization of these phase-change emulsions are largely unexplored. Here, the acoustic vaporization of individual micron-sized perfluoropentane (PFP) droplets is studied at a nanoseconds timescale. Highly diluted emulsions of PFP-in-water and oil-in-PFP-in-water droplets, ranging from 3.5 to 11 µm in radius, were prepared and the nucleation and growth of the vapor bubbles was imaged at frame rates of up to 20 Mfps. The droplet vaporization dynamics was observed to have three distinct regimes: (1) prior to nucleation, a regime of droplet deformation and oscillatory translations within the surrounding fluid along the propagation direction of the applied ultrasound; (2) a regime characterized by the rapid growth of a vapor bubble enhanced by ultrasound-driven rectified heat transfer; and (3) a final phase characterized by a relatively slow expansion, after ultrasound stops, that is fully dominated by heat transfer. A method to measure the moment of inception of the nucleation event with respect to the phase of the ultrasound wave is proposed. A simple physical model captures quantitatively all of the features of the subsequent vapor bubble growth.

Safety of ultrasound contrast agents in patients with known or suspected cardiac shunts.

Contrast-enhanced ultrasound imaging is a radiation-free diagnostic tool that uses biocompatible ultrasound contrast agents (UCAs) to improve image clarity. UCAs, which do not contain dye, often salvage "technically difficult" ultrasound scans, increasing the accuracy and reliability of a front-line ultrasound diagnosis, reducing unnecessary downstream testing, lowering overall health care costs, changing therapy, and improving patient care. Two UCAs currently are approved and regulated by the US Food and Drug Administration. They have favorable safety profiles and risk/benefit ratios in adult and pediatric populations, including compromised patients with severe cardiovascular diseases. Nevertheless, these UCAs are contraindicated in patients with known or suspected right-to-left, bidirectional, or transient right-to-left cardiac shunts. These patients, who constitute 10% to 35% of the general population, typically receive no UCAs when they undergo echocardiography. If their echocardiographic images are suboptimal, they may receive inappropriate diagnosis and treatment, or they may be referred for additional diagnostic testing, including radiation-based procedures that increase their lifetime risk for cancer or procedures that use contrast agents containing dye, which may increase the risk for kidney damage. An exhaustive review of current peer-reviewed research demonstrated no scientific basis for the UCA contraindication in patients with known or suspected cardiac shunts. Initial safety concerns were based on limited rodent data and speculation related to macroaggregated albumin microspheres, a radioactive nuclear imaging agent with different physical and chemical properties and no relation to UCAs. Radioactive macroaggregated albumin is not contraindicated in adult or pediatric patients with cardiac shunts and is routinely used in these populations. In conclusion, the International Contrast Ultrasound Society Board recommends removal of the contraindication to further the public interest in safe, reliable, radiation-free diagnostic imaging options for patients with known or suspected cardiac shunts and to reduce their need for unnecessary downstream testing.

Acoustic Droplet Vaporization for the Enhancement of Ultrasound Thermal Therapy.

Acoustic droplet vaporization (ADV) is an ultrasound method for converting biocompatible microdroplets into microbubbles. The objective is to demonstrate that ADV bubbles can enhance high intensity focused ultrasound (HIFU) therapy by controlling and increasing energy absorption at the focus. Thermal phantoms were made with or without droplets. Compound lesions were formed in the phantoms by 5-second exposures with 5-second delays. Center to center spacing of individual lesions was 5.5 mm in either a linear pattern or a spiral pattern. Prior to the HIFU, 10 cycle tone bursts with 0.25% duty cycle were used to vaporize the droplets, forming an "acoustic trench" within 30 seconds. The transducer was then focused in the middle of the back bubble wall to form thermal lesions in the trench. All lesions were imaged optically and with 2T MRI. With the use of ADV and the acoustic trench, a uniform thermal ablation volume of 15 cm(3) was achieved in 4 minutes; without ADV only less than 15% of this volume was filled. The commonly seen tadpole shape characteristic of bubble-enhanced HIFU lesions was not evident with the acoustic trench. In conclusion, ADV shows promise for the spatial control and dramatic acceleration of thermal lesion production by HIFU.

Functional imaging with intraoperative ultrasound: detection of somatosensory cortex in dogs with color-duplex sonography.

OBJECTIVE: To evaluate the capability of intraoperative color-duplex sonography to detect eloquent flow-activated areas and their anatomic relationship in dogs. METHODS: After craniotomy, the sensory cortex of eight dogs was identified by recording the highest amplitude detected with a grid electrode evoked with somatosensory evoked potential stimulation of the nervus ischiadicus. A 7.5-MHz linear array transducer was placed on the dura, and eight images were taken in color-coded capture mode during baseline and somatosensory evoked potential stimulation of the ipsilateral (nonevoked) and contralateral (evoked) sensory cortex. The differences in flow velocity intensities were statistically compared (Wilcoxon test) in three arbitrary velocity ranges and across all colored pixels in a region of interest between baseline and stimulation in both hemispheres. RESULTS: Comparing both hemispheres during stimulation, the evoked sensory cortex demonstrated an increase of 10% in the number of counted colored pixels during stimulation, whereas the number of counted colored pixels in the ipsilateral sensory cortex decreased by 2% (P < 0.05), indicating an overall increase in measured flow during stimulation. Comparing differences during nonstimulation and stimulation in single hemispheres, the lowest of the three velocity ranges (approximately 10-20 mm/s) demonstrated a statistically significant (P = 0.01) increase during stimulation, whereas no change was observed during stimulation in the ipsilateral hemisphere. This increase has been confirmed by regional cerebral blood flow measurement with colored microspheres. CONCLUSION: This study indicates, for the first time, the capability of intraoperative ultrasound to detect functionally important areas during evoked stimulation.