Ultrasound-mediated Drug Delivery Research Articles

Clinical Applications of Micro/Nanobubble Technology in Neurological Diseases.

Nanomedicine, leveraging the unique properties of nanoparticles, has revolutionized the diagnosis and treatment of neurological diseases. Among various nanotechnological advancements, ultrasound-mediated drug delivery using micro- and nanobubbles offers promising solutions to overcome the blood-brain barrier (BBB), enhancing the precision and efficacy of therapeutic interventions. This review explores the principles, current clinical applications, challenges, and future directions of ultrasound-mediated drug delivery systems in treating stroke, brain tumors, neurodegenerative diseases, and neuroinflammatory disorders. Additionally, ongoing clinical trials and potential advancements in this field are discussed, providing a comprehensive overview of the impact of nanomedicine on neurological diseases.

Real-time non-invasive control of ultrasound hyperthermia using high-frequency ultrasonic backscattered energy in ex vivo tissue and in vivo animal studies

Objective.A reliable, calibrated, non-invasive thermometry method is essential for thermal therapies to monitor and control the treatment. Ultrasound (US) is an effective thermometry modality due to its relatively high sensitivity to temperature changes, and fast data acquisition and processing capabilities.Approach.In this work, the change in backscattered energy (CBE) was used to control the tissue temperature non-invasively using a real-time proportional-integral-derivative (PID) controller. A clinical high-frequency US scanner was used to acquire radio-frequency echo data fromex vivoporcine tissue samples andin vivomice hind leg tissue while the tissue was treated with mild hyperthermia by a focused US applicator. The PID controller maintained the focal temperature at approximately 40 °C for about 4 min.Main results.The results show that the US thermometry based on CBE estimated by a high-frequency US scanner can produce 2D temperature maps of a localized heating region and to estimate the focal temperature during mild hyperthermia treatments. The CBE estimated temperature varied by an average of ±0.85 °C and ±0.97 °C, compared to a calibrated thermocouple, inex vivoandin vivostudies, respectively. The mean absolute deviations of CBE thermometry during the controlled hyperthermia treatment were ±0.45 °C and ±0.54 °C inex vivoandin vivo,respectively.Significance.It is concluded that non-invasive US thermometry via backscattered energies at high frequencies can be used for real-time monitoring and control of hyperthermia treatments with acceptable accuracy. This provides a foundation for an US mediated drug delivery system.

Ultrasound-Mediated Antibiotic Delivery to In Vivo Biofilm Infections: A Review.

Bacterial biofilms are a significant concern in various medical contexts due to their resilience to our immune system as well as antibiotic therapy. Biofilms often require surgical removal and frequently lead to recurrent or chronic infections. Therefore, there is an urgent need for improved strategies to treat biofilm infections. Ultrasound-mediated drug delivery is a technique that combines ultrasound application, often with the administration of acoustically-active agents, to enhance drug delivery to specific target tissues or cells within the body. This method involves using ultrasound waves to assist in the transportation or activation of medications, improving their penetration, distribution, and efficacy at the desired site. The advantages of ultrasound-mediated drug delivery include targeted and localized delivery, reduced systemic side effects, and improved efficacy of the drug at lower doses. This review scrutinizes recent advances in the application of ultrasound-mediated drug delivery for treating biofilm infections, focusing on in vivo studies. We examine the strengths and limitations of this technology in the context of wound infections, device-associated infections, lung infections and abscesses, and discuss current gaps in knowledge and clinical translation considerations.

Numerical analysis of ultrasound-mediated microbubble interactions in vascular systems: Effects on shear stress and vessel mechanics

The present study concerns the numerical modeling of microbubble oscillation within an elastic microvessel, aiming to enhance the safety and efficacy of ultrasound-mediated drug delivery and diagnostic imaging. The success of such applications depends on a thorough understanding of microbubble–vessel interactions. Despite some progress, the critical impact of the stabilizing shell around gas core has remained underexplored. To address this, we developed a novel numerical approach that models the stabilizing shell. Additionally, there is novelty in modeling consequent vascular deformation in response to complicated spatiotemporal microbubble oscillations. The novel approach was implemented for shear stress evaluation as a critical factor in vascular permeability. Finally, our unique approach offered novel insights into microbubble–vessel interactions under diverse acoustic conditions. Results indicated substantial impact of shell properties and acoustic parameters on induced shear stress. With a fourfold increase in acoustic pressure amplitude, 15.6-fold and sixfold increases were observed in maximum shear stress at 1 and 3 MHz, respectively. Also, the peak shear stress could reach up to 15.6 kPa for a shell elasticity of 0.2 N/m at 2.5 MHz. Furthermore, decreasing microvessel/bubble size ratio from 3 to 1.5 increased maximum shear stress from 5.1 to 24.3 kPa. These findings are crucial for optimizing ultrasound parameters in clinical applications, potentially improving treatment outcomes while minimizing risk of vessel damage. However, while our model demonstrated high fidelity in reproducing experimental observations, it is limited by assumptions of vessel geometry and homogeneity of vessel properties. Future work can improve our findings through in vitro experimental measurements.

Evaluation of Doxorubicin-loaded Echogenic Macroemulsion for Targeted Drug Delivery.

The latest technology trend in targeted drug delivery highlights stimuliresponsive particles that can release an anticancer drug in a solid tumor by responding to external stimuli. This study aims to design, fabricate, and evaluate an ultrasound-responsive drug delivery vehicle for an ultrasound-mediated drug delivery system. The drug-containing echogenic macroemulsion (eME) was fabricated by an emulsification method using the three phases (aqueous lipid solution as a shell, doxorubicin (DOX) contained oil, and perfluorohexane (PFH) as an ultrasound-responsive agent). The morphological structure of eMEs was investigated using fluorescence microscopy, and the size distribution was analyzed by using DLS. The echogenicity of eME was measured using a contrast-enhanced ultrasound device. The cytotoxicity was evaluated using a breast cancer cell (MDA-MB-231) via an in vitro cell experiment. The obtained eME showed an ideal morphological structure that contained both DOX and PFH in a single particle and indicated a suitable size for enhancing ultrasound response and avoiding complications in the blood vessel. The echogenicity of eME was demonstrated via an in vitro experiment, with results showcasing the potential for targeted drug delivery. Compared to free DOX, enhanced cytotoxicity and improved drug delivery efficiency in a cancer cell were proven by using DOX-loaded eMEs and ultrasound. This study established a platform technology to fabricate the ultrasound-responsive vehicle. The designed drug-loaded eME could be a promising platform with ultrasound technology for targeted drug delivery.

Fabrication of porous poly(L-lactide-co-ε-caprolactone) micropowder for microbubble effect and ultrasound-mediated drug delivery.

Biodegradable elastic poly(L-lactide-co-ε-caprolactone) (PLCL) copolymer (50:50, lactide:caprolactone molar ratio) was synthesized and porous PLCL micropowders was fabricated by a simple method involving rapid cooling of 0.1, 0.5, and 1% (wt/vol) PLCL/dioxane spray into liquid nitrogen. The physicochemical properties of the porous PLCL micropowders were examined by measuring their pore size, pore morphology, and microbead size using a scanning electron microscopy (SEM) and dye and temozolomide (TMZ)-release testing under ultrasound. Human U-87MG, glioblastoma (GBM) cell culture tests were performed to evaluate cell cytotoxicity by released drug from PLCL micropowders. In this study, the porous PLCL micropowders prepared from 1 wt%/vol% PLCL solutions showed a highly porous structure, satisfactory mechanical properties, and optimal drug release efficiency compared with those produced from 0.1 or 0.5 wt%/vol% solutions. The results of the accumulated release test with the results of the absorbance of the dye initially applied, it was confirmed that more than 80% of the added dye was trapped inside the micropowder, and clearly GBM cytotoxicity effect could be observed by the released TMZ. The drug release system using micropowders and ultrasound can be applied as a drug supply system for various diseases such as brain tumors with low drug permeability.

RGD-coated polymeric microbubbles promote ultrasound-mediated drug delivery in an inflamed endothelium-pericyte co-culture model of the blood-brain barrier

Drug delivery to central nervous pathologies is compromised by the blood-brain barrier (BBB). A clinically explored strategy to promote drug delivery across the BBB is sonopermeation, which relies on the combined use of ultrasound (US) and microbubbles (MB) to induce temporally and spatially controlled opening of the BBB. We developed an advanced in vitro BBB model to study the impact of sonopermeation on the delivery of the prototypic polymeric drug carrier pHPMA as a larger molecule and the small molecule antiviral drug ribavirin. This was done under standard and under inflammatory conditions, employing both untargeted and RGD peptide-coated MB. The BBB model is based on human cerebral capillary endothelial cells and human placental pericytes, which are co-cultivated in transwell inserts and which present with proper transendothelial electrical resistance (TEER). Sonopermeation induced a significant decrease in TEER values and facilitated the trans-BBB delivery of fluorescently labeled pHPMA (Atto488-pHPMA). To study drug delivery under inflamed endothelial conditions, which are typical for e.g. tumors, neurodegenerative diseases and CNS infections, tumor necrosis factor (TNF) was employed to induce inflammation in the BBB model. RGD-coated MB bound to and permeabilized the inflamed endothelium-pericyte co-culture model, and potently improved Atto488-pHPMA and ribavirin delivery. Taken together, our work combines in vitro BBB bioengineering with MB-mediated drug delivery enhancement, thereby providing a framework for future studies on optimization of US-mediated drug delivery to the brain.Graphical abstract

Real time cavitation monitoring during ultrasound-mediated drug delivery in normothermically perfused human tumour-bearing livers

In the fight against a broad spectrum of human diseases, cavitation techniques show great promise for overcoming physical barriers that lead to suboptimal uptake of passively administered therapeutics. However, small animal testing of candidate therapies remains a poor predictor of clinical success. Here we demonstrate ultrasound-mediated drug delivery in normal and tumour-bearing human livers infused with protein-based cavitation nuclei (PCaN). Whole and partial human livers were obtained immediately from hepatectomy surgeries and were normothermically sustained using a clinically approved perfusion system (OrganOx Metra). Ultrasound was applied using a 0.5 MHz focused source (Sonic Concepts H107) and was monitored with a calibrated linear array (ATS L7-4) for real time structural and cavitational imaging implemented on an array controller (Verasonics Vantage 256). Specifically, the therapy process was monitored using passive acoustic mapping (PAM) of broadband cavitation emissions, employing a non-adaptive beamformer that deconvolves the array point spread function. Levels of fluorescently labelled drugs incorporated in and co-administered with the PCaN were quantified in blood and tissue samples collected during and following treatment, respectively. This presentation highlights PAM observations of broadband cavitation persistence and drug delivery in untargeted and targeted tissues, including the first ever experiments in tumour-bearing human livers.

A mathematical model for maximizing drug uptake in low-EPR tumors using different methods of ultrasound-mediated drug delivery

Cancer remains a significant health burden and research efforts aim to improve drug delivery options. Focused ultrasound has emerged as a potential mechanism for enhanced targeted drug delivery in tumors with a low Enhanced Permeability and Retention (EPR) effect by “sonoporation”. Traditional sonoporation has shown promise, but it can still benefit from improvements in drug uptake. The current study investigates the use of a new non-portative technique, called “ultrasonic tumor painting,” to enhance vascular retention of nanoparticles in the tumor space to achieve targeted drug delivery and increase therapeutic efficacy. An existing mathematical model was adapted to simulate the concentration of free and liposome encapsulated doxorubicin in circulation. The simulations were performed in MATLAB and specifically evaluate doxorubicin uptake in low EPR tumors. To increase drug uptake, sonoporation was employed where efficiency of intratumoral liposomal doxorubicin uptake was increased. An alternative non-permeabilizing strategy, called “tumor painting,” was utilized to improve vascular retention instead of increasing permeability. After retention, the small molecule doxorubicin is allowed to leak into the tumors space via concentration mediated Fickian Diffusion. This study demonstrates that ultrasound-based drug delivery can be improved with tumor painting. This method is a valuable tool to overcome the limitations of traditional sonoporation and has exciting potential for enhancing the therapeutic efficacy of chemotherapy.

The Evolution and Recent Trends in Acoustic Targeting of Encapsulated Drugs to Solid Tumors: Strategies beyond Sonoporation.

Despite recent advancements in ultrasound-mediated drug delivery and the remarkable success observed in pre-clinical studies, no delivery platform utilizing ultrasound contrast agents has yet received FDA approval. The sonoporation effect was a game-changing discovery with a promising future in clinical settings. Various clinical trials are underway to assess sonoporation's efficacy in treating solid tumors; however, there are disagreements on its applicability to the broader population due to long-term safety issues. In this review, we first discuss how acoustic targeting of drugs gained importance in cancer pharmaceutics. Then, we discuss ultrasound-targeting strategies that have been less explored yet hold a promising future. We aim to shed light on recent innovations in ultrasound-based drug delivery including newer designs of ultrasound-sensitive particles specifically tailored for pharmaceutical usage.

Abstract 4304: Network-based inference identifies cell state-specific drugs targeting master regulator vulnerabilities in diffuse midline glioma

Abstract Diffuse Midline Glioma (DMG) are fatal pediatric brain tumors with no therapies. We leveraged network-based methodologies to dissect the heterogeneity of DMG tumors and to discover Master Regulator (MR) proteins representing pharmacologically accessible, mechanistic determinants of molecularly distinct cell states. We produced the first DMG regulatory network from 122 publicly available RNAseq profiles with ARACNe (Basso et al. Nat Genet 2005), and inferred sample-specific MR protein activity with VIPER (Alvarez et al. Nat Genet 2016) based on the differential expression of their targets. 7 of the top 25 most active MRs found comprise a well-characterized MR block (MRB2) (Paull et al.Cell 2021), frequently activated across aggressive tumors, and enriched in DMG patient MR signatures (Fisher’s Exact Test p=4.4 × 10−18). A CRISPR/Cas9 KO screen across 3 DMG patient cell lines identified a set of 73/77 essential genes that were enriched in the MR signature of 80% of patient samples (GSEA p=0.000034). FOXM1 emerged as an essential MR, significantly activated across virtually all patients. We then generated RNAseq profiles following perturbation with ~300 oncology drugs in 2 DMG cell lines most representative of patient MR signatures, and used this to identify drugs that invert patient MR activity profiles using the NYS/CA Dept. of Health approved OncoTreat algorithm (Alvarez et al. Nat Genet 2018). OncoTreat predicted sensitivity to HDAC, MEK, CDK, PI3K, and proteosome inhibitors in subsets of patients, overlapping with published DMG drug screens. Importantly, 80% of OncoTreat-predicted drugs (p<10−5) from 3 DMG patient tumor biopsies showed in vitro sensitivity in cultured tumor cells from the respective patients, with overall 68% accuracy among 223 drugs evaluated by both OncoTreat and in vitro (Fisher’s Exact Test p=0.0449). Further analysis of DMG intra-tumor heterogeneity via protein activity inference across DMG single cells from 6 published scRNAseq profiles identified 6 tumor clusters with unique MR signatures co-existing in virtually all patients representing distinct cellular states (2 astrocyte-, 1 oligodendrocyte-, and 3 oligodendrocyte precursor cell-like states). Targetable MRs and OncoTreat-predicted drugs were distinct between these states. Bulk RNAseq analysis recapitulated predictions seen in the more prevalent OPC-like states, but failed to capture MR and drug predictions for the AC-like states (e.g. JAK1/Ruxolitinib and STAT3/Napabucasin). We are currently validating cell state-specific drug predictions in vivo at single-cell resolution in subcutaneous patient-derived xenograft and orthotopic syngeneic DMG models that we have shown recapitulate patient tumor heterogeneity, including with focused ultrasound-mediated drug delivery. This provides a platform to nominate much-needed novel drugs and drug combinations to treat DMG. Citation Format: Ester Calvo Fernandez, Junqiang Wang, Xu Zhang, Hong-Jian Wei, Hanna E. Minns, Aaron T. Griffin, Lukas Vlahos, Timothy J. Martins, Pamela S. Becker, John Crawford, Robyn D. Gartrell, Luca Szalontay, Stergios Zacharoulis, Zhiguo Zhang, Robert Wechsler-Reya, Cheng-Chia Wu, Andrea Califano, Jovana Pavisic. Network-based inference identifies cell state-specific drugs targeting master regulator vulnerabilities in diffuse midline glioma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4304.

Drug delivery approaches for enhanced antibiofilm therapy.

Biofilms have attracted increasing attention in recent years. Many bacterial infections are associated with biofilm formation. A bacterial biofilm is an aggregated membrane-like substance that is composed of a large number of bacteria and their secreted extracellular polymeric substances. The traditional antibiofilm approaches, such as chemotherapy based on antibiotics, are often ineffective in eradicating biofilms owing to the limited diffusion ability of antibiotics within biofilms and inactivation of antibiotics by biofilms. Moreover, a larger dosage of antibiotics could be effective, but leads to an increased tolerance. Smart drug delivery systems that deliver antibiotics into the biofilm interior is a promising strategy to meet this challenge. In this review, we focus on the methods to improve drug delivery efficiency for enhanced chemotherapy of biofilms. Furthermore, we have summarized chemical approaches for enhanced drug delivery, such as chemical shields, charge reversal, and dual corona enhanced delivery strategies; these methods focus on physicochemical biofilm properties and specific biofilm features. Afterwards, physical approaches are discussed, such as magnetism-mediated drug delivery, electricity-mediated drug delivery, ultrasound-mediated drug delivery, and shock wave-mediated drug delivery. Finally, a perspective on the development of next-generation antibiofilm drug delivery systems is given.

A REVIEW ARTICLE ON THE NOVEL MECHANISMS FOR DELIVERY OF CANCER THERAPEUTICS

The majority of cancer treatments used today focus on surgically removing the tumour mass. The rapid proliferation of malignant cells is significantly slowed down by physical and chemical therapies like radiotherapy and chemotherapy. Additionally, these treatments have a high level of toxicity and severe side effects, which decrease patient compliance. To improve treatment indices, new approaches to conventional cancer therapeutics, including cancer stem cell therapy, triggered release, intracellular drug targeting, gene delivery, magnetic drug targeting, ultrasound mediated drug delivery and photodynamic therapy have added new techniques for treating cancer. These methods have aided in the selective detection of cancerous cells, enabling their eradication with the fewest possible negative effects. This review discusses cutting-edge methods for administering chemotherapy more effectively in an effort to improve cancer prognosis. KEYWORDS - cancer stem cells, photodynamic therapy, gene therapy, triggered release

Effect of acoustic cluster therapy (ACT®) combined with chemotherapy in a patient-derived xenograft mouse model of pancreatic cancer.

Pancreatic ductal adenocarcinomas respond poorly to chemotherapy, in part due to the dense tumor stroma that hinders drug delivery. Ultrasound (US) in combination with microbubbles has previously shown promise as a means to improve drug delivery, and the therapeutic efficacy of ultrasound-mediated drug delivery is currently being evaluated in multiple clinical trials. However, most of these utilize echogenic contrast agents engineered for imaging, which might not be optimal compared to specialized formulations tailored for drug delivery. In this study, we evaluated the in vivo efficacy of phase-shifting microbubble-microdroplet clusters that, upon insonation, form bubbles in the size range of 20-30μm. We developed a patient-derived xenograft model of pancreatic cancer implanted in mice that largely retained the stromal content of the originating tumor and compared tumor growth in mice given chemotherapeutics (nab-paclitaxel plus gemcitabine or liposomal irinotecan) with mice given the same chemotherapeutics in addition to ultrasound and acoustic cluster therapy. We found that acoustic cluster therapy significantly improved the effect of both chemotherapeutic regimens and resulted in 7.2 times higher odds of complete remission of the tumor compared to the chemotherapeutics alone.

Ultrasound-mediated delivery of flexibility-tunable polymer drug conjugates for treating glioblastoma.

Effective chemotherapy delivery for glioblastoma multiforme (GBM) is limited by drug transport across the blood-brain barrier and poor efficacy of single agents. Polymer-drug conjugates can be used to deliver drug combinations with a ratiometric dosing. However, the behaviors and effectiveness of this system have never been well investigated in GBM models. Here, we report flexible conjugates of hyaluronic acid (HA) with camptothecin (CPT) and doxorubicin (DOX) delivered into the brain using focused ultrasound (FUS). In vitro toxicity assays reveal that DOX-CPT exhibited synergistic action against GBM in a ratio-dependent manner when delivered as HA conjugates. FUS is employed to improve penetration of DOX-HA-CPT conjugates into the brain in vivo in a murine GBM model. Small-angle x-ray scattering characterizations of the conjugates show that the DOX:CPT ratio affects the polymer chain flexibility. Conjugates with the highest flexibility yield the highest efficacy in treating mouse GBM in vivo. Our results demonstrate the association of FUS-enhanced delivery of combination chemotherapy and the drug-ratio-dependent flexibility of the HA conjugates. Drug ratio in the polymer nanocomplex may thus be employed as a key factor to modulate FUS drug delivery efficiency via controlling the polymer flexibility. Our characterizations also highlight the significance of understanding the flexibility of drug carriers in ultrasound-mediated drug delivery systems.

Microbubble-Nanoparticle Complexes for Ultrasound-Enhanced Cargo Delivery.

Targeted delivery of therapeutics to specific tissues is critically important for reducing systemic toxicity and optimizing therapeutic efficacy, especially in the case of cytotoxic drugs. Many strategies currently exist for targeting systemically administered drugs, and ultrasound-controlled targeting is a rapidly advancing strategy for externally-stimulated drug delivery. In this non-invasive method, ultrasound waves penetrate through tissue and stimulate gas-filled microbubbles, resulting in bubble rupture and biophysical effects that power delivery of attached cargo to surrounding cells. Drug delivery capabilities from ultrasound-sensitive microbubbles are greatly expanded when nanocarrier particles are attached to the bubble surface, and cargo loading is determined by the physicochemical properties of the nanoparticles. This review serves to highlight and discuss current microbubble-nanoparticle complex component materials and designs for ultrasound-mediated drug delivery. Nanocarriers that have been complexed with microbubbles for drug delivery include lipid-based, polymeric, lipid-polymer hybrid, protein, and inorganic nanoparticles. Several schemes exist for linking nanoparticles to microbubbles for efficient nanoparticle delivery, including biotin-avidin bridging, electrostatic bonding, and covalent linkages. When compared to unstimulated delivery, ultrasound-mediated cargo delivery enables enhanced cell uptake and accumulation of cargo in target organs and can result in improved therapeutic outcomes. These ultrasound-responsive delivery complexes can also be designed to facilitate other methods of targeting, including bioactive targeting ligands and responsivity to light or magnetic fields, and multi-level targeting can enhance therapeutic efficacy. Microbubble-nanoparticle complexes present a versatile platform for controlled drug delivery via ultrasound, allowing for enhanced tissue penetration and minimally invasive therapy. Future perspectives for application of this platform are also discussed in this review.

284 (PB064) - Ultrasound-induced cavitation enhances therapeutic efficacy of AXL-targeting ADC leading to improved survival in a human xenograft model of renal cancer

Background: Antibody-drug conjugates (ADCs) are a new class of antitumor agents that combine the targeting specificity of monoclonal antibodies (mAbs) with the cell killing ability of powerful cytotoxic molecules. However, poor penetration of drugs into solid tumors, including mAbs, is a well-recognized barrier to effective therapy as this strongly limits the number of tumor cells targeted by the therapeutic agent. A novel non-invasive approach to tackle this problem is ultrasound-mediated drug delivery using co-infused cavitation nuclei, which shows promise as a way to pump more drug (including mAbs/ADCs) into and throughout tumors. The purpose of this study was to test a suboptimal single dose of mipasetamab uzoptirine (ADCT-601), an ADC targeting the AXL protein, in combination with ultrasound-induced cavitation in a preclinical mouse model of renal cancer. Material and methods: SCID mice were injected subcutaneously with SN12C human renal cancer cells and when tumors reached an average size of 150 mm3, animals were divided into 6 cohorts (N = 6–7 mice each): a) PBS (vehicle control); b-c) B12-PL1601 (control ADC) at 0.3 mg/kg ± cavitation; d–e) ADCT-601 at 0.3 mg/kg ± cavitation; f) ADCT-601 at 1 mg/kg (efficacious dose). Cavitation nuclei and drug agent were administered intravenously just before ultrasound application from the OxSonics SonoCart system (ultrasound parameters: fc = 0.5 MHz; 8000cycles; PRF = 0.5 Hz; PRFP = 1–2.9 MPa variable). Ultrasound amplitude was set according to the cavitation monitoring method (passive acoustic mapping [PAM]) to be within 0.1–0.5 nJ/pulse for 10 minutes of treatment. Mice were culled when tumors’ width + length passed the value of 24 mm. Results: Results indicated that cavitation was achieved in the desired range for all ultrasound groups. Following treatment, 2 groups (ADCT-601 at 1 mg/kg and ADCT-601 at 0.3 mg/kg + cavitation) showed significant tumor growth delay, ultimately leading to a significant improvement in survival when compared to all other groups. No significant difference between the high and the low ADCT-601 dose with cavitation was observed, indicating that the combination enhanced potency by 3.3-fold. Conclusions: Cavitation-enhanced delivery of ADCT-601 has a strong effect on xenograft tumor growth leading to a significant increase in survival. This was demonstrated for a suboptimal ADC dose, indicating that ultrasound-mediated drug delivery could be used to reduce the administered dose while preserving therapeutic efficacy. These preclinical data warrant further evaluation of this promising combination in the clinic. Conflict of interest: Ownership: Massimo Masiero, Erika Vojtasova, Paul Boulos, Calum Crake, Christian Coviello and Cliff Rowe own OxSonics Therapeutics share options. Christian Coviello also owns OxSonics Therapeutics shares. Francesca Zammarchi and Patrick van Berkel own ADC Therapeutics shares. Other Substantive Relationships: Massimo Masiero, Erika Vojtasova, Paul Boulos, Calum Crake, Christian Coviello and Cliff Rowe are OxSonics Therapeutics employees. Christian Coviello is also OxSonics Therapeutics co-founder. Francesca Zammarchi and Patrick van Berkel are ADC Therapeutics employees.

Acoustic radiation force dependence on properties of elastic spherical shells in standing waves

In this study, we theoretically investigate the acoustic radiation force acting on elastic spherical shells in standing waves in the dimensionless frequency range of 0<ka<0.7 (k is the wave number in the host fluid and a is the outer radius of the spherical shells), we show that in the low-frequency range (0<ka<0.2), the acoustic radiation force on core–shell spheres can vary from positive to negative by decreasing the effective density and effective bulk modulus of the spherical shells, while the effective density and bulk modulus are dependent on the hollow ratio of the core–shell spheres, the density and bulk modulus ratios of the shell to the core. Thus, the core–shell spheres cannot only be trapped from the pressure node to the pressure antinode by increasing the hollow ratio or decreasing the shell’s density or longitudinal or shear velocity, but are also unresponsive to ultrasound waves with an optimal hollow ratio, density, or acoustic velocity of the shell. We further show that in the relatively low-frequency range of0<ka<0.7, the acoustic radiation force on core–shell spheres can be significantly enhanced around resonant frequencies. The acoustic radiation force could be enhanced at lower frequencies by increasing the hollow ratio or decreasing the shell’s density or shear wave velocity. Our results pave the way for optimising the design of core–shell particles for efficient ultrasound-mediated drug delivery.

Evaluation of the potential of ultrasound-mediated drug delivery for the treatment of ovarian cancer through preclinical studies

Ovarian cancer (OC) has the greatest mortality rate among gynecological cancers, with a five-year survival rate of <50%. Contemporary adjuvant chemotherapy mostly fails in the case of OCs that are refractory, metastatic, recurrent, and drug-resistant. Emerging ultrasound (US)-mediated technologies show remarkable promise in overcoming these challenges. Absorption of US waves by the tissue results in the generation of heat due to its thermal effect causing increased diffusion of drugs from the carriers and triggering sonoporation by increasing the permeability of the cancer cells. Certain frequencies of US waves could also produce a cavitation effect on drug-filled microbubbles (MBs, phospholipid bilayers) thereby generating shear force and acoustic streaming that could assist drug release from the MBs, and promote the permeability of the cell membrane. A new class of nanoparticles that carry therapeutic agents and are guided by US contrast agents for precision delivery to the site of the ovarian tumor has been developed. Phase-shifting of nanoparticles by US sonication has also been engineered to enhance the drug delivery to the ovarian tumor site. These technologies have been used for targeting the ovarian cancer stem cells and protein moieties that are particularly elevated in OCs including luteinizing hormone-releasing hormone, folic acid receptor, and vascular endothelial growth factor. When compared to healthy ovarian tissue, the homeostatic parameters at the tissue microenvironment including pH, oxygen levels, and glucose metabolism differ significantly in ovarian tumors. US-based technologies have been developed to take advantage of these tumor-specific alterations for precision drug delivery. Preclinical efficacy of US-based targeting of currently used clinical chemotherapies presented in this review has the potential for rapid human translation, especially for formulations that use all substances that are deemed to be generally safe by the U.S. Food and Drug Administration.

Synthesis of echogenic liposomes for sonoporation

Ultrasound contrast agents (UCAs), which are groups of engineered microbubbles, have been recently studied for drug delivery applications, since the cavitation of bubbles can increase the temporary permeability of nearby cells. However, the internal volume of UCAs is generally filled with gas, hence loading drug molecules into UCAs is limited. In this study, an echogenic liposome with a liquid and gas core is proposed as an alternative carrier of genetic material for ultrasound-mediated drug delivery. The structure of the synthesized echogenic liposome was analysed via transmission electron microscopy and confocal microscopy with fluorescent labels. The protection of siRNA by the echogenic liposomes was also verified by exposure to RNase. The results indicate that at least 10% of the total siRNA used in the experiment was successfully protected by the proposed echogenic liposome. Additionally, the release of siRNA from the liposomes could be successfully achieved with 1 W/cm2 ultrasound sonication at 1 MHz; parameters low enough to be used in generic ultrasound therapeutic systems. Although further studies to clarify the responses to incident ultrasound fields and to quantitatively analyse the internal liquid volume for drug loading are required, the proposed echogenic liposome has great potential for ultrasound-mediated gene delivery.