Lipid Microbubbles Research Articles

Ultrasound-Targeted Microbubble Destruction Accelerates Angiogenesis and Ameliorates Left Ventricular Dysfunction after Myocardial Infarction in Mice

Failure of coronary recanalization within 12 h or no flow in the myocardium after percutaneous coronary intervention is associated with high mortality from myocardial infarction, and insufficient angiogenesis in the border zone results in the expansion of infarct area. In this study, we examined the effects of ultrasound-targeted microbubble destruction (UTMD) on angiogenesis and left ventricular dysfunction in a mouse model of myocardial infarction. Fifty-four mice with MI were treated with no UTMD, ultrasound (US) alone or UTMD four times (days 1, 3, 5 and 7), and another 18 mice underwent sham operation and therapy. Therapeutic US was generated with a linear transducer connected to a commercial diagnostic US system (VINNO70). UTMD was performed with the VINNO70 at a peak negative pressure of 0.8 MPa and lipid microbubbles. Transthoracic echocardiography was performed on the first and seventh days. The results indicated that UTMD decreased the infarct size ratio from 78.1 ± 5.3% (untreated) to 43.3 ± 6.4%, accelerated angiogenesis and ameliorated left ventricular dysfunction. The ejection fraction increased from 25.05 ± 8.52% (untreated) to 42.83 ± 9.44% (UTMD). Compared with that in other groups, expression of vascular endothelial growth factor and endothelial nitric oxide synthase and release of nitric oxide were significantly upregulated after UTMD treatment, indicating angiogenesis. Therefore, UTMD is a potential physical approach in the treatment of myocardial infarction.

Temperature controlled transformations of giant unilamellar vesicles of amphiphilic triblock copolymers synthesized via microfluidic mixing

We report on a simple approach for synthesis of temperature-responsive giant unilamellar vesicles (GUVs) from poly(N-vinylcaprolactam)15-block-poly(dimethylsiloxane)65-block-poly(N-vinylcaprolactam)15 (PVCL15-PDMS65-PVCL15) triblock copolymer and non-temperature responsive small and giant vesicles from novel poly(N-vinylpyrrolidone)-block-poly(dimethylsiloxane)-block-poly(N-vinylpyrrolidone) (PVPON15-PDMS65-PVPON15 and PVPON6-PDMS30-PVPON6) triblock copolymers using microfluidic mixing at 25 °C. We show that temperature-responsive PVCL15-PDMS65-PVCL15 GUVs with the average diameter of 1.4 ± 0.2 µm while being stable at room temperature for at least 14 days, transformed irreversibly into small vesicles of 168 ± 40 nm after incubation of their aqueous solution at 42 °C for 24 h. We hypothesized that this transformation is induced by local compressive stresses of the vesicle membrane due to the collapse of PVCL blocks above the copolymer lower critical solution temperature (LCST) leading to the decrease of the vesicle membrane thickness. Consequently, we found that the temperature-induced size transformation of the PVCL-based GUVs at 42°C can be suppressed by substituting PVCL with its hydrophilic homologue PVPON, or by suppressing the PVCL's LCST behavior through hydrogen-bonding with tannic acid molecules. In the former case, novel PVPONn-PDMSm-PVPONn triblock copolymers (n = 15, m = 65 and n = 6, m = 30) assemble into vesicles stable from 25 °C to 55 °C as confirmed by optical and electron microscopy, dynamic light scattering (DLS) and small-angle neutron scattering (SANS). In the latter, hydrogen bonding interactions of PVCL with the polyphenol tannic acid (TA) at room temperature resulted in stable PVCL-based GUVs at 42°C as confirmed by optical, electron, and atomic force microscopies. We also found that physical crosslinking of the PVCL corona through hydrogen bonding with TA in PVCL15-PDMS65-PVCL15 GUVs will delay their low pH-induced degradation at 37°C by 48 h compared to non-modified GUVs. Our findings open opportunities for the development of temperature-regulated stable micro-vehicles that would change their structural characteristics in the physiologically relevant temperature range from 25 to 42°C and can be utilized for cell mimicking studies. The developed GUVs also have potential in theranostic drug delivery as substitutes for polymer microcapsules and lipid microbubbles as well as for stimuli-triggered sensing, protection, and rapid response in an aqueous environment.

Sonoporation Using Nanoparticle-Loaded Microbubbles Increases Cellular Uptake of Nanoparticles Compared to Co-Incubation of Nanoparticles and Microbubbles.

Therapeutic agents can benefit from encapsulation in nanoparticles, due to improved pharmacokinetics and biodistribution, protection from degradation, increased cellular uptake and sustained release. Microbubbles in combination with ultrasound have been shown to improve the delivery of nanoparticles and drugs to tumors and across the blood-brain barrier. Here, we evaluate two different microbubbles for enhancing the delivery of polymeric nanoparticles to cells in vitro: a commercially available lipid microbubble (Sonazoid) and a microbubble with a shell composed of protein and nanoparticles. Various ultrasound parameters are applied and confocal microscopy is employed to image cellular uptake. Ultrasound enhanced cellular uptake depending on the pressure and duty cycle. The responsible mechanisms are probably sonoporation and sonoprinting, followed by uptake, and to a smaller degree enhanced endocytosis. The use of commercial Sonazoid microbubbles leads to significantly lower uptake than when using nanoparticle-loaded microbubbles, suggesting that proximity between cells, nanoparticles and microbubbles is important, and that mainly nanoparticles in the shell are taken up, rather than free nanoparticles in solution.

AFGF Targeted Mediated by Novel Nanoparticles-Microbubble Complex Combined With Ultrasound-Targeted Microbubble Destruction attenuates Doxorubicin-Induced Heart Failure via Anti-Apoptosis and Promoting Cardiac Angiogenesis.

The purpose of this study was to evaluate the protective effect of acidic fibroblast growth factor targeted mediated by novel nanoparticles–cationic lipid microbubbles complex (aFGF–NP + CPMBs) combined with ultrasound targeted microbubble destruction (UTMD)on doxorubicin–induced heart failure (HF)and its mechanism. Heart failure rats induced by intraperitoneal injection with doxorubicin (DOX) to achieve cummulative dose of 15mg/kg for continuous 6 weeks showed left ventricular dysfunction, seriously oxidative stress, cardiomyocyte apoptosis, and decrease of myocardial vascular density. In contrast, aFGF–NP + CPMBs combined with UTMD therapy (3ug/kg, caudal vein injection, twice a week, 6weeks)prominently ameliorated left ventricular dysfunction by increased ejection fraction (EF) and fractional shortening (FS), decreased brain natriuretic peptide (BNP); strengthened the ability of antioxidant stress confirmed by increasing the activity of SOD and reducing the production of MDA; exerted the effect of anti–cardiomyocyte apoptosis and promotion angiogenesis by inhibited Bax expression and increased Bcl–2 expression and platelet endothelial cell adhesion molecule (CD31) expression. Taken together, the research suggested that aFGF targeted mediated by novel nanoparticles–cationic lipid microbubbles complex combined with UTMD should be a promising targeted treatment for heart failure.

Production of Membrane-Filtered Phase-Shift Decafluorobutane Nanodroplets from Preformed Microbubbles.

There are many methods that can be used for the production of vaporizable phase-shift droplets for imaging and therapy. Each method utilizes different techniques and varies in price, materials, and purpose. Many of these fabrication methods result in polydisperse populations with non-uniform activation thresholds. Additionally, controlling the droplet sizes typically requires stable perfluorocarbon liquids with high activation thresholds that are not practical in vivo. Producing uniform droplet sizes using low-boiling point gases would be beneficial for in vivo imaging and therapy experiments. This article describes a simple and economical method for the formation of size-filtered lipid-stabilized phase-shift nanodroplets with low-boiling point decafluorobutane (DFB). A common method of generating lipid microbubbles is described, in addition to a novel method of condensing them with high-pressure extrusion in a single step. This method is designed to save time, maximize efficiency, and generate larger volumes of microbubble and nanodroplet solutions for a wide variety of applications using common laboratory equipment found in many biological laboratories.

Ultrasound Microbubble-Mediated VHL Regulates the Biological Behavior of Ovarian Cancer Cells

According to the literature, the von Hippel–Lindau (VHL) gene has a certain correlation with ovarian cancer. In this study, we investigated the effect and mechanism of ultrasound microbubble-mediated VHL on the biological function of ovarian cancer cells. Non-targeting lipid microbubbles and targeted lipid microbubbles were prepared. OVCAR-3 cells were treated with VHL mediated by ultrasound and microbubbles alone or together. Expressions of VHL, Akt, epithelial–mesenchymal-transition-related proteins and apoptosis-related proteins were detected by Western blot and quantitative real-time polymerase chain reaction as needed. The effect of ultrasound microbubble-mediated VHL on the proliferation, apoptosis, cell cycle, migration and invasion of OVCAR-3 cells was examined by Cell Counting Kit-8, flow cytometry, wound-healing assay and Transwell. Compared with other treatment methods, ultrasound microbubble mediation enhanced VHL expression in OVCAR-3 cells. Overexpression of liposome-mediated VHL inhibited the proliferation and migration; caused cell-cycle arrest; promoted apoptosis: downregulated the expressions of MMP2, MMP9, E-cadherin, Akt and Bcl-2; and upregulated the expressions of VHL and BCL2-associated X protein (BAX) in OVCAR-3 cells. The effect of microbubble-treated VHL was similar to liposome-mediated regulation, while ultrasound treatment enhanced the effect of VHL on OVCAR-3 cells. More interestingly, ultrasound microbubble-mediated VHL had the most obvious regulatory effect on OVCAR-3 cells. Ultrasound microbubble technology increases the transfection efficiency of VHL into OVCAR-3 cells and enhances the effect of VHL gene on the biological function of OVCAR-3 cells.

STAT3 decoy oligonucleotide-carrying microbubbles with pulsed ultrasound for enhanced therapeutic effect in head and neck tumors.

Signal transducer and activator of transcription-3 (STAT3) is an oncogenic transcription factor implicated in carcinogenesis, tumor progression, and drug resistance in head and neck squamous cell carcinoma (HNSCC). A decoy oligonucleotide targeting STAT3 offers a promising anti-tumor strategy, but achieving targeted tumor delivery of the decoy with systemic administration poses a significant challenge. We previously showed the potential for STAT3 decoy-loaded microbubbles, in conjunction with ultrasound targeted microbubble cavitation (UTMC), to decrease tumor growth in murine squamous cell carcinoma. As a next step towards clinical translation, we sought to determine the anti-tumor efficacy of our STAT3 decoy delivery platform against human HNSCC and the effect of higher STAT3 decoy microbubble loading on tumor cell inhibition. STAT3 decoy was loaded on cationic lipid microbubbles (STAT3-MB) or loaded on liposome-conjugated lipid microbubbles to form STAT3-loaded liposome-microbubble complexes (STAT3-LPX). UTMC treatment efficacy with these two formulations was evaluated in vitro using viability and apoptosis assays in CAL33 (human HNSCC) cells. Anti-cancer efficacy in vivo was performed in a CAL33 tumor murine xenograft model. UTMC with STAT3-MB caused significantly lower CAL33 cell viability compared to UTMC with STAT3-LPX (56.8±8.4% vs 84.5±8.8%, respectively, p<0.05). In vivo, UTMC with STAT3-MB had strong anti-tumor effects, with significantly less tumor burden and greater survival compared to that of UTMC with microbubbles loaded with a mutant control decoy and untreated control groups (p<0.05). UTMC with STAT3 decoy-loaded microbubbles significantly decreases human HNSSC tumor progression. These data set the stage for clinical translation of our microbubble platform as an imaged-guided, targeted delivery strategy for STAT3 decoy, or other nucleotide-based therapeutics, in human cancer treatment.

Ultrasound Imaging Based on Magnetic Lipid Microbubble Contrast Agent with Fe₃O₄ Nanoparticles.

To construct magnetic microbubbles with dual response characteristics of ultrasound and magnetic field. The surfactant-based microbubble ultrasound contrast agent (ST68) was prepared using acoustic cavitation. Magnetic Fe₃O₄ nanoparticles having a negative charge on the surface were prepared by the polyol method. The microbubbles were used as the template, and the magnetic microbubbles were prepared by alternating deposition of polyethyleneimine and magnetic Fe₃O₄ nanoparticles on the surface of microbubbles using electrostatic attraction for a layer-by-layer selfassembly. An in vitro ultrasound contrast device was set up to compare the ultrasound images of the silicone tube in the device before and after the injection of human magnetic microbubbles (3×10/M), and to observe the movement of the magnetic microbubbles after the magnetic field was applied to the silicone tube. To evaluate the contrast effect of magnetic microbubbles in vivo, we compared the renal ultrasound images of New Zealand white rabbits before and after injection of human magnetic microbubbles. The surface of Fe₃O₄ nanoparticles displayed a stable negative charge (-24.6±67 mV). More than 98% of the assembled magnetic microbubbles were smaller than 8 μm, which met the basic size requirement for an ultrasound contrast agent. There was no echo signal in the silicone tube before the injection of magnetic microbubbles. Following injection, a solid echo was detected in the silicone tube. After the application of the magnetic field, the magnetic microbubbles migrated in the direction of magnetic field. Contrast-enhanced ultrasound did not image the kidney before the injection of magnetic microbubbles, while the kidney shadow was clear following injection. Magnetic targeted microbubbles and contrast-enhanced ultrasound provide good imaging, which lays the foundation for further studies of magnetic targeted microbubble contrast agents in diagnosis and treatment.

Ultrasonic features and pathological information of lymphoepithelial carcinomas of the salivary gland with ferroferric oxide nanometer ultrasound contrast agent

This study aimed to investigate and analyze the ultrasonic features and pathological information of lymphoepithelial carcinomas of the salivary gland using ultrasound. The general clinical information of 62 patients with lymphoepithelial carcinomas of the salivary gland and the ultrasonic features and pathological information of 28 patients who underwent surgery were retrospectively analyzed and compared with ferroferric oxide (Fe3O4) nanometer lipid microbubble contrast agent using Minitab software with P &lt; 0.005 to indicate statistical significance. Similarly, inflammatory nodules, lymphomas, and metastatic nasopharyngeal carcinomas were identified using ultrasound. Lymphoepithelial carcinomas were often observed in the parotid glands, with hypoechoic nodules. Network-like or cord-like hyperechoes were often observed in the nodules, with unclear boundaries, irregular morphologies, and significantly less invasion into the surrounding tissues. Differences in the incidence in the right and left glands, blood flow signal, nerve invasion, lymphatic metastasis, and tumor volume were insignificant. Lymphatic metastasis was often noted in ipsilateral neck level II, followed by the periparotid region or ipsilateral neck level I. The diagnoses of inflammatory nodules and lymphomas had a combined pathology, while the diagnosis of nasopharynx cancer was dependent on the result of nasopharyngoscopy. The ultrasonic features of lymphoepithelial carcinomas of the salivary gland are network-like or cord-like hypoechoic nodules that possess the ultrasonic and biological features of malignant tumors.

Lipid Microbubble-Conjugated Anti-CD3 and Anti-CD28 Antibodies (Microbubble-Based Human T Cell Activator) Offer Superior Long-Term Expansion of Human Naive T Cells In Vitro.

Stimulation of human primary T cells with immobilized anti-CD3 and anti-CD28 Abs in vitro provide a system to study T cell activation and proliferation and an avenue for expanding T cells for immunotherapy. Magnetic beads conjugated with anti-CD3 and anti-CD28 Abs (Dynabeads Human T-Activator [D-TCA]) have been a golden standard for stimulating human primary T cells in vitro. In this study, we report that an application using anti-CD3 and anti-CD28 Abs conjugated on lipid microbubbles (microbubble-based human T cell activator [MB-TCA]) to stimulate primary human naive T cells resulted in expansion superior to D-TCA. In 56-d cultures with three repeated stimulation cycles (14 d per stimulation), we found that 1) MB-TCA induced significantly better expansion (20- and 10-fold increase) of naive CD4+ and CD8+ T cells than did D-TCA; 2) MB-TCA- and D-TCA-stimulated T cells had a similar number of initial cell divisions, but MB-TCA had significantly lower activation-induced cell death than D-TCA; 3) MB-TCA-stimulated T cells produced less TNF-α than did D-TCA; and 4) blocking TNF-α action via adding an Ab against TNF-αR (TNFRSF1A) significantly improved expansion of T cells activated by D-TCA in vitro. Together, we demonstrated that the MB-TCA induces a better expansion of human naive T cells in vitro and offers advantages in both basic and clinical applications in which the outcome depends on the number of T cells.

Ultrasound-responsive lipid microbubbles for drug delivery: A review of preparation techniques to optimise formulation size, stability and drug loading.

Lipid-shelled microbubbles have received extensive interest to enhance ultrasound-responsive drug delivery outcomes due to their high biocompatibility. While therapeutic effectiveness of microbubbles is well established, there remain limitations in sample homogeneity, stability profile and drug loading properties which restrict these formulations from seeing widespread use in the clinical setting. In this review, we evaluate and discuss the most encouraging leads in lipid microbubble design and optimisation. We examine current applications in drug delivery for the systems and subsequently detail shell compositions and preparation strategies that improve monodispersity while retaining ultrasound responsiveness. We review how excipients and storage techniques help maximise stability and introduce different characterisation and drug loading techniques and evaluate their impact on formulation performance. The review concludes with current quality control measures in place to ensure lipid microbubbles can be reproducibly used in drug delivery.

Anti-CD3 and anti-CD28 antibodies conjugated with lipid microbubbles (MB-TAC) provide a superior expansion of human naive CD4+ and CD8+ T cells in long term culture

Abstract Anti-CD3 and anti-CD28 antibodies are a method of choice for general and physiological activation of human primary T cells in vitro. Here we report a comparison of the growth, cell death, and cytokine production of anti-CD3 and anti-CD28 antibodies conjugated with lipid microbubbles (MB-TCA) and the standard Dynal beads (D-TCA). In 56-day cultures with three repeated stimulation cycles (14 days per stimulation), we found that MB-TCA induced a significantly better expansion (~20-fold increase) of naïve CD4 and CD8 T cells than did D-TCA. We found the main difference between MB-TCA and D-TCA was not the speed of activation-induced initial T cell proliferation but rather the survival of activated T cells. Furthermore, we found that MB-TCA activated T cells had a better telomere length maintenance and produced less TNFα and other cytokines than did D-TCA. Finally, blocking TNFα action via antibody against TNFRSF1A significantly improved expansion of T cells activated by D-TCA in vitro. Together, we demonstrated that MB-TCA offers a better expansion of primary human T cells in vitro and will be a useful method for long term culture of primary human T cells for both basic and clinical applications.

One-Step Microfluidic Coating of Phospholipid Microbubbles with Natural Alginate Polymer as a Delivery System for Human Epithelial Lung Adenocarcinoma.

In this study, the neoplastic drug frequently used in the treatment of lung cancer, carboplatin is loaded to microbubbles via a microfluidic platform. In order to increase the drug loading capacity of microbubbles, carboplatin is encapsulated into alginate polymer layer. The phospholipid microbubbles (MBs) are synthesized by MicroSphere Creator, which is connected with T-junction and micromixer for the treatment with CaCl2 solution to provide gelation of the alginate coated phospholipid microbubbles (AMBs). The carboplatin loaded alginate coated phospholipid microbubbles (CAMBs) result in 12.2 ± 0.21 µm mean size, obtained by mixing with 0.05% CaCl2 using T-junction. The cytotoxic activities of the synthesized MBs, AMBs, and CAMBs are also investigated with the 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) and live/dead fluorescent dying assays in the A549 and BEAS-2B cell lines. The one-step microfluidic coating of lipid microbubbles with natural alginate polymer appears to be a promising strategy for enhanced drug reservoir properties.

Ultrasound-Mediated Co-Delivery of miR-34a and sPD-1 Complexed with Microbubbles for Synergistic Cancer Therapy.

BackgroundmiR-34a was downregulated and PD-L1 was upregulated in cervical cancer; however, the treatment of cervical cancer lacks precision and targeting. This study explored the ultrasound-mediated co-delivery of miR-34a and sPD-1 complexes with microbubbles for synergistic cancer therapy.MethodsCationic lipid microbubbles (CLMBs) were prepared by membrane hydration and mechanical oscillation. U14 subcutaneous xenograft mice were injected with CLMBs-loaded sPD-1 and miR-34a combined with ultrasound targeted destruction, and tumor volume and tumor weight of mice were measured. TUNEL apoptosis test and the mRNA expression of apoptosis-related gene Bcl-2 and Bax were analyzed by qRT-PCR. Antitumor immune-related cytokines IFN-γ were investigated by qRT-PCR, LDH Cytotoxicity Assay Kit were performed to test cytotoxic T lymphocytes (CTL).ResultsCLMBs were successfully prepared and the plasmid bound to its surface. The tumor volume and weight were specifically decreased by ultrasound-mediated co-delivery of miR-34a and sPD-1 complexes with microbubbles, apoptosis was induced and the apoptosis suppressor gene Bcl-2 was downregulated and proapoptotic gene Bax were upregulated. qRT-PCR analysis revealed that antitumor immunity-related IFN-γ was strongly upregulated in mice, which were treated with CLMBs-loaded sPD-1 and miR-34a combined with ultrasound targeted destruction, and the percentage of CTL was increased.ConclusionThese findings from the study demonstrated that CLMBs could deliver miR-34a and sPD-1, combined with ultrasound targeted destruction, could suppress the tumor tissue growing, induce apoptosis and enhance antitumor immunity in U14 subcutaneous xenograft mice.

Ultrasound Responsive Magnetic Mesoporous Silica Nanoparticle-Loaded Microbubbles for Efficient Gene Delivery

Purpose: Gene therapy is an important therapeutic strategy for cancer. Nanoparticles are used for noninvasive gene delivery, which has great potential in tumor therapy. However, it is a challenge to construct a targeted gene delivery vector with high gene delivery efficiency, good biocompatibility, and multiple functions. Method: Herein, we designed magnetic mesoporous silica nanoparticle loading microbubbles (M-MSN@MBs) for ultrasound-mediated imaging and gene transfection. The plasmid DNA (pDNA) was encapsulated into the pores of M-MSNs. Also, the pDNA-carrying M-MSNs were loaded in the lipid microbubbles. Results: The gene vector presented good biocompatibility, DNA binding stability, ultrasound imaging performance, and magnetic responsiveness. The polyethyleneimine (PEI)-modified M-MSNs effectively protected the loaded pDNA from enzyme degradation. The cytotoxicity of M-MSNs was significantly reduced via encapsulating in lipid microbubbles. Upon the magnetic field, M-MSN@MBs were attracted to the tumor area. Then, ultrasound-targeted microbubble destruction (UTMD) not only released loaded M-MSNs but also facilitated M-MSNs delivery to tumor tissue by opening blood-tumor barrier and increasing the cytomembrane permeability, and ultimately improved the pDNA delivery efficiency. Conclusion: Our findings suggested that the developed ultrasound-responsive gene delivery system was a promising platform for gene therapy, which could noninvasively enhance tumor gene transfection.

3:18 PM Abstract No. 229 Antitumor effect of transarterial chemoembolization using doxorubicin-albumin nanoparticle loaded lipid microbubbles combined with ultrasound-targeted activation on VX2 rabbit liver tumors

To explore the antitumor effect of transarterial chemoembolization using Doxorubicin-albumin nanoparticle loaded lipid microbubbles (mbTACE) combined with ultrasound-targeted microbubble activation (US) in rabbit VX2 liver tumor model. VX2 rabbits were assigned to 4 groups (n = 7/group): (i) saline (control); (ii) conventional TACE (cTACE); (iii) mbTACE without ultrasound activation (US-); (iv) mbTACE with ultrasound activation (US+). To assess hepatic damage, serum aspartate transaminase and alanine transaminase levels were measured on baseline and 1,3,and 7 days posttherapy. Changes of tumor volumes were segmented on MRI (baseline, 7 days posttherapy). To assess tumor necrosis, animals were euthanized on day 7, and explanted tumors were stained with hematoxylin and eosin and a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling assay. Percentage areas of viable tumors were calculated using digitalized histopathologic specimen images. estimated viable tumor volume was calculated using viable tumor fraction multiplying tumor volume on day 7 MRI. Treated groups show significant tumor suppression effect on MR compared to control group, without significant inter-treatment group difference. Tumor viable fraction and viable volume were 5.34% ± 6.54% and 89.9 mm3 ± 91.73 mm3, 17.28% ± 9.18% and 643.98 mm3 ± 758.15 mm3, 12.37% ± 8.11% and 187.01 mm3 ± 100.79 mm3, and 35.27% ± 6.46% and 1778.167 mm3± 1512.82 mm3in groups US+mbTACE, US-mbTACE, cTACE, and control (P < .001 and P < .001). Liver enzyme levels were elevated after drug delivery but recovered during follow-up. No significant differences were observed between treated groups on days 1, 3, and 7. Treatment with a combination of ultrasound activation and mbTACE showed therapeutic benefit in a liver tumor animal model.

Spectral Imaging for Microbubble Characterization.

Microbubbles stabilized by an outer lipid shell have been studied extensively for both diagnostic and therapeutic applications. The shell composition can significantly influence microbubble behavior, but performing quantitative measurements of shell properties is challenging. The aim of this study is to investigate the use of spectral imaging to characterize the surface properties of a range of microbubble formulations representing both commercial and research agents. A lipophilic dye, C-laurdan, whose fluorescence emission varies according to the properties of the local environment, was used to compare the degree and uniformity of the lipid order in the microbubble shell, and these measurements were compared with the acoustic response and stability of the different formulations. Spectral imaging was found to be suitable for performing rapid and hence relatively high throughput measurements of microbubble surface properties. Interestingly, despite significant differences in lipid molecule size and charge, all of the different formulations exhibited highly ordered lipid shells. Measurements of liposomes with the same composition and the debris generated by destroying lipid microbubbles with ultrasound (US) showed that these exhibited a lower and more varied lipid order than intact microbubbles. This suggests that the high lipid order of microbubbles is due primarily to compression of the shell as a result of surface tension and is only minimally affected by composition. This also explains the similarity in acoustic response observedbetween the formulations, because microbubble dynamics are determined by the diameter and shell viscoelastic properties that are themselves a function of the lipid order. Within each population, there was considerable variability in the lipid order and response between individual microbubbles, suggesting the need for improved manufacturing techniques. In addition, the difference in the lipid order between the shell and lipid debris may be important for therapeutic applications in which shedding of the shell material is exploited, for example, drug delivery.

Effect of Temperature on Stability of Lipid Microbubbles

The effect of temperature on stability of lipid microbubble shell containing polyethyleneoxide-40-stearate (PEG40St) as emulsifier was investigated. Microbubbles at 4 °C were subjected to different temperatures up to 48 ºC (down-to-up) and it was found that both the number and the size of microbubbles remained unchanged in the population up to a certain time, so called “onset time”. The onset time was about 6 hrs at 10 °C, 2 hrs at 20 °C and shorter at elevated temperatures, exhibiting an exponential decrease with increasing temperature. Once the onset time was reached, the number of microbubbles started to decrease and the average size of the population started to increase. Observation of single microbubbles on a constant temperature heating stage exhibited that each microbubble had its own onset time, with the smaller microbubbles vanishing earlier than the larger ones. The Langmuir monolayer studies showed that hydration degree of the emulsifier PEG chains decreased with temperature, causing them go through conformational changes and subsequently destabilization of the shell. By subjecting the freshly produced microbubbles directly to the desired temperatures in up-to-down fashion, more stable microbubbles were able to be produced, with their onset time increased 40% at 10 °C to 500% at 38 °C. Overall, the results suggest that the new strategies need to be developed to control the collapse process in the microbubble shell resulting from the conformational changes in the PEG chains of the emulsifier for the design of more stable microbubbles.

Horses for courses (or something much smaller!): The design, development and acoustic testing of sub-micron cavitation nucleation agents for biomedical applications

Lipid and protein-shelled microbubbles have been the mainstay of cavitation nucleation strategies for therapeutic applications for several decades. However, a growing number of therapeutic ultrasound applications require nuclei that (i) are significantly smaller in size, in order to overcome a particular biological barrier such as the leaky vasculature of tumours or the stratum corneum; (ii) offer greatly increased cavitation persistence, both during a single extended ultrasound pulse and in terms of extended circulation following intravenous administration; and (iii) have better resilience to sudden ambient pressure changes in order to enable direct injection without nuclei destruction into tissue targets via a needle and syringe. We will review a range of novel cavitation agents currently under development, including sub-micron gas-stabilizing solid particles and nanodroplets, and provide an overview of their known characteristics in terms of acoustic emissions, activation and cavitation thresholds, cavitation persistence, and circulation. Where possible, the relevant performance of these cavitation nucleation agents will be compared to microbubbles for applications ranging from drug delivery to nucleated tissue fractionation.

Low intensity ultrasound targeted microbubble destruction assists MSCs delivery and improves neural function in brain ischaemic rats

Background and purpose: The present study aimed to explore the feasibility and efficacy of the targeted non-invasive implantation of mesenchymal stromal cells (MSCs) by low-intensity ultrasound-targeted microbubble destruction (LI-UTMD) assisted blood–brain barrier (BBB) opening and its improvement on neurobehavioural outcomes in brain ischaemic rats.Methods: A transcranial irradiation of low-intensity ultrasound by diagnostic devices was performed, and lipid microbubbles (MBs) and MSCs were simultaneously infused. Then, the MSC transmigration from brain vessels to parenchyma was demonstrated, and MSCs were statistically analysed on days 1, 4, 7 and 14. Behavioural function was statistically analysed.Results: The extra-vascular leakage of lanthanum and EB was observed at the brain ischaemic area receiving ultrasound. MSCs were observed at the ultrasound irradiated brain hemisphere, and the number of MSCs in LI-UTMD assisted MSCs group was significantly higher than that in the MSCs group (p < .01). The attachment, traversing and trans-migration of MSCs across the BBB were recorded. Neuro-behavioural function was improved with this approach.Conclusions: The transcranial irradiation of low intensity ultrasound targeted MBs destruction on brain ischaemic rats might be a safe and efficient BBB opening approach to prompt the successful delivery of MSCs into the targeted area of brain ischaemia, and ameliorate neurological function.