Ultrasound 96 Probe Device Protocol for cancer cell treatment v1

Ultrasound is a sound wave with frequencies ranging between 20 kHz and 20 MHz. Ultrasound is able to temporarily and repeatedly open the BBB safely and enhance chemotherapeutic delivery without adverse effects.(Deprez et al., 2021). This novel technique in drug delivery benefits from the powerful ability of ultrasound to produce cavitation activity. Cavitation is the generation and activity of gas-filled bubbles in a medium exposed to ultrasound. As the pressure wave passes through the media, gas bubbles expand at low pressure and contract at high pressure. This leads to oscillation which produces a circulating fluid flow known as microstreaming around the bubble with velocities and shear rates proportional to the amplitude of the oscillation. At high amplitudes the associated shear forces can cut open liposomes (Wanigasekara et al., 2021; Deprez et al., 2021). Vesicles denser than the surrounding liquid are drawn into the shear field surrounding an oscillating bubble. If the shear stress is greater than the strength of the vesicle, it will burst and spill its contents. In a liposome, the vesicle will reform, often at a smaller size than before meeting the shear field. Hence, some interior liquid must be released during the break down. (Pitt et al., 2004) This protocol describes the use of an ultrasound probe to trigger the release of liposomes in glioblastoma cells. This method uses an ultrasound device which is set to the following parameters: Time = 3 min, Pulse = 59 /01, Amplitude = 20%. The ultrasound technique is an easy and reliable technique making it useful in the study of a variety of areas such as oncology. When applied to an ultrasonic transducer, the Pulser part of the instrument generates short, large amplitude electric pulses of controlled energy, which are transformed into short ultrasonic pulses. The VCX 750 is the ultrasonic liquid processor used for this experiment. It is powerful and versatile and can process a wide range of sample types and volumes for many different applications.

Efficiency of ultrasonic treatment of polysaccharide from brown algae

Ultrasonic exposure can be used for depolymerization of brown algae polysaccharides. However, its effectiveness depends on several factors, including cavitation activity in the treatment medium. Therefore, the purpose of the work was to determine the cavitation activity and the effectiveness of the ultrasonic exposure to fucoidan in order to optimize the processing processes of polysaccharide from brown algae. A change in cavitation activity was revealed depending on the composition of the processing environment, as well as on the intensity of ultrasonic exposure with a constant frequency of the ultrasonic wave. Similar dynamics of change of cavitation activity were established at the intensity of ultrasonic treatment of 100 and 133 W/cm2 with amplification of electric signal at the increase of ultrasound intensity. The use of SDS in the processing medium led to an increase in cavitation activity to 14.9±0.47 mV. Treatment of the fucoidan solution for 40 minutes under various conditions allowed to obtain fractions with a change in the average hydrodynamic particle diameter from 113 nm (100 W/cm2) to 85 nm (200 W/cm2) and 124 nm (SDS).

Gas-stabilizing nanoparticles for ultrasound imaging and therapy of cancer

The use of ultrasound in the clinic has been long established for cancer detection and image-guided tissue biopsies. In addition, ultrasound-based methods have been widely explored to develop more effective cancer therapies such as localized drug delivery, sonodynamic therapy, and focused ultrasound surgery. Stabilized fluorocarbon microbubbles have been in use as contrast agents for ultrasound imaging in the clinic for several decades. It is also known that microbubble cavitation could generate thermal, mechanical, and chemical effects in the tissue to improve ultrasound-based therapies. However, the large size, poor stability, and short-term cavitation activity of microbubbles limit their applications in cancer imaging and therapy. This review will focus on an alternative type of ultrasound responsive material; gas-stabilizing nanoparticles, which can address the limitations of microbubbles with their nanoscale size, robustness, and high cavitation activity. This review will be of interest to researchers who wish to explore new agents to develop improved methods for molecular ultrasound imaging and therapy of cancer.

Acoustic frequency-dependent physical mechanism of sub-MHz ultrasound neurostimulation

Ultrasound is an innovative physical modality allowing non-invasive and reversible modulation of neural circuit activity in the brain with high spatial resolution. Despite growing interest in clinical applications, the safe and effective use of ultrasound neuromodulation has been limited by a lack of understanding of the physical mechanisms underlying its effects. Here, we demonstrate acoustic frequency-dependent physical effects that underlie ultrasound neuromodulation, where cavitation and radiation forces are the dominant sources of low- and high-frequency stimulation, respectively. We used 39.5 kHz and 500 kHz acoustic frequencies to stimulate cultured neural and glial cells, excised from rat cortex, to study acoustic frequency-dependent neural responses. We demonstrate increased evoked responses due to increased cavitation activity at the 39.5 kHz acoustic frequency. In contrast, notable cavitation activity was not detected at 500 kHz despite detection of evoked responses. Our work highlights the dependence of ultrasound neuromodulation on acoustic frequencies, with different physical effects underlying cell responses to low and high sub-MHz acoustic frequency ranges.

Synergistic Cisplatin-induced cell death by ultrasound-microbubble mediated intracellular delivery in breast cancer cells.

Ultrasound-microbubble (USMB) potentiated cisplatin (CDDP) therapy was assessed in human breast cancer cells. Cells, MDA-MB-231, in suspension were exposed to USMB and CDDP at varying conditions, during which microbubble cavitation activity was measured using passive cavitation detection and 48 hours post-treatment cell viability and intracellular platinum concentration were measured using MTT assay and mass cytometry, respectively. USMB synergistically enhanced cell death (~20 fold) when combined with CDDP and significantly increased intracellular CDDP concentration (~8 fold) compared to CDDP treatment alone. Cell death and intracellular CDDP concentration were correlated to microbubble cavitation activity, which increased with peak negative pressure and microbubble concentration. Combined treatment of USMB and CDDP at relatively lower integrated cavitation dose (ICD) induced a synergistic effect on cell death whereas ICD greater than 10 induced an additive effect. USMB mediated CDDP intracellular accumulation synergistically enhances cell death in CDDPresistant breast cancer cells.

Synergistic Cisplatin-induced cell death by ultrasound-microbubble mediated intracellular delivery in breast cancer cells.

Ultrasound-microbubble (USMB) potentiated cisplatin (CDDP) therapy was assessed in human breast cancer cells. Cells, MDA-MB-231, in suspension were exposed to USMB and CDDP at varying conditions, during which microbubble cavitation activity was measured using passive cavitation detection and 48 hours post-treatment cell viability and intracellular platinum concentration were measured using MTT assay and mass cytometry, respectively. USMB synergistically enhanced cell death (~20 fold) when combined with CDDP and significantly increased intracellular CDDP concentration (~8 fold) compared to CDDP treatment alone. Cell death and intracellular CDDP concentration were correlated to microbubble cavitation activity, which increased with peak negative pressure and microbubble concentration. Combined treatment of USMB and CDDP at relatively lower integrated cavitation dose (ICD) induced a synergistic effect on cell death whereas ICD greater than 10 induced an additive effect. USMB mediated CDDP intracellular accumulation synergistically enhances cell death in CDDPresistant breast cancer cells.

Dynamics of cavitation zone development during sonication of suspensions of magnesium particles

The cavitation activity during ultrasonic treatment of magnesium particles has been investigated. The cavitation activity recorded in a continuous mode of ultrasonic treatment altered in a wide range at constant output parameters of the generator. The rate and nature of cavitation activity variation depended on the mass fraction of particles in the suspension. It has been demonstrated that during the ultrasonic treatment of magnesium aqueous suspensions it is possible to determine the following stages: growth of cavitation activity, reaching a maximum followed by a decrease and reaching a plateau (or repeated cycles of increasing or decreasing cavitation activity). The complex nature of the cavitation activity dynamics is associated with the participation of hydrogen released as a result of the chemical interaction of magnesium particles with water in the formation of the cavitation zone. The magnesium particles modified with ultrasound were characterised with the use of scanning electron microscopy, X-ray phase analysis and thermal analysis. It has been found that ultrasonic treatment of magnesium particles resulted in the formation of magnesium hydroxide and magnesium hydride phases.