Pickering Emulsions and Antibubbles Stabilized by PLA/PLGA Nanoparticles.

Micrometer-sized double emulsions and antibubbles were produced and stabilized via the Pickering mechanism by colloidal interfacial layers of polymeric nanoparticles (NPs). Two types of nanoparticles, consisting either of polylactic acid (PLA) or polylactic-co-glycolic acid (PLGA), were synthesized by the antisolvent technique without requiring any surfactant. PLA nanoparticles were able to stabilize water-in-oil (W/O) emulsions only after tuning the hydrophobicity by means of a thermal treatment. A water-in-oil-in-water (W/O/W) emulsion was realized by emulsifying the previous W/O emulsion in a continuous water phase containing hydrophilic PLGA nanoparticles. Both inner and outer water phases contained a sugar capable of forming a glassy phase, while the oil was crystallizable upon freezing. Freeze drying the double emulsion allowed removing the oil and water and replacing them with air without losing the three-dimensional (3D) structure of the original emulsion owing to the sugar glassy phase. Reconstitution of the freeze-dried double emulsion in water yielded a dispersion of antibubbles, i.e., micrometric bubbles containing aqueous droplets, with the interfaces of the antibubbles being stabilized by a layer of adsorbed polymeric nanoparticles. Remarkably, it was possible to achieve controlled release of a flourescent probe (calcein) from the antibubbles through heating to 37 °C leading to bursting of the antibubbles.

Generation of antibubbles from core-shell double emulsion templates produced by microfluidics.

We report the preparation of antibubbles by microfluidic methods. More specifically, we demonstrate a two-step approach, wherein a monodisperse water-in-oil-in-water (W/O/W) emulsion of core-shell construction is first generated via microfluidics and freeze-dried thereafter to yield, upon subsequent reconstitution, an aqueous dispersion of antibubbles. Stable antibubbles are attained by stabilization of the air-water interfaces through a combination of adsorbed particles and polymeric surfactant. The antibubbles strongly resemble the double emulsion templates from which they were formed. When triggered to release, antibubbles show complete release of their cores within about 100 ms.

Sono-processes: Emerging systems and their applicability within the (bio-)medical field.

Sonochemistry, although established in various fields, is still an emerging field finding new effects of ultrasound on chemical systems and are of particular interest for the biomedical field. This interdisciplinary area of research explores the use of acoustic waves with frequencies ranging from 20 kHz to 1 MHz to induce physical and chemical changes. By subjecting liquids to ultrasonic waves, sonochemistry has demonstrated the ability to accelerate reaction rates, alter chemical reaction pathways, and change physical properties of the system while operating under mild reaction conditions. It has found its way into diverse industries including food processing, pharmaceuticals, material science, and environmental remediation. This review provides an overview of the principles, advancements, and applications of sonochemistry with a particular focus on the domain of (bio-)medicine. Despite the numerous benefits sonochemistry has to offer, most of the research in the (bio-)medical field remains in the laboratory stage. Translation of these systems into clinical practice is complex as parameters used for medical ultrasound are limited and toxic side effects must be minimized in order to meet regulatory approval. However, directing attention towards the applicability of the system in clinical practice from the early stages of research holds significant potential to further amplify the role of sonochemistry in clinical applications.

Antibubbles Enable Tunable Payload Release with Low-Intensity Ultrasound.

The benefits of ultrasound are its ease-of-use and its ability to precisely deliver energy in opaque and complex media. However, most materials responsive to ultrasound show a weak response, requiring the use of high powers, which are associated with undesirable streaming, cavitation, or temperature rise. These effects hinder response control and may even cause damage to the medium where the ultrasound is applied. Moreover, materials that are currently in use rely on all-or-nothing effects, limiting the ability to fine-tune the response of the material on the fly. For these reasons, there is a need for materials that can respond to low intensity ultrasound with programmable responses. Here it is demonstrated that antibubbles are a low-intensity-ultrasound-responsive material system that can controllably release a payload using acoustic pressures in the kilopascal range. Varying their size and composition tunes the release pressure, and the response can be switched between a single release and stepwise release across multiple ultrasound pulses. Observations using confocal and high-speed microscopy reveal different ways that can lead to release. These findings lay the groundwork to design antibubbles that controllably respond to low-intensity ultrasound, opening a wide range of applications ranging from ultrasound-responsive material systems to carriers for targeted delivery.

High-Throughput Fabrication of Size-Controlled Pickering Emulsions, Colloidosomes, and Air-Coated Particles via Clog-Free Jetting of Suspensions.

Microcapsules with a liquid core and a solid shell composed of hydrophobic nanoparticles are broadly applied in food, pharmaceutics, and biotechnologies. For example, Pickering emulsions, colloidosomes, or antibubbles (droplets surrounded by air layers in water) enable controlled release of active agents, biocompatibility, and contact-less liquid transportation. However, producing controlled nanoparticle- or polymer-laden hydrophobic shells at scale is highly challenging, since bulk methods are polydisperse and microfluidic chips are prone to clogging and slow. Here, clog-free coating of an aqueous jet with silica nanoparticle suspensions with concentrations up to 10% (w/v), as well as high concentrations of polymers (30% (w/v) poly(lactic acid) (PLA)), is demonstrated, enabling continuous generation of microcapsules at flow rates up to 4 mL min-1 . Pickering emulsions are converted into capsules, providing hydrophobic shells consisting of nanoparticles for controlled release. As a highlight, the scalable fabrication of air-coated capsules (antibubbles) in the sub-millimeter range is demonstrated. The shell contains an air film that protects the liquid core for days yet enables ultrasound-induced release within 3 min. By enabling rapid fabrication of controlled Pickering emulsions, colloidosomes, antibubbles, and biodegradable capsules, jetting through a liquid layer (JetALL) provides a versatile platform for advanced applications in food, pharmacy, and life science.

Preparation of acid-responsive antibubbles from CaCO3-based Pickering emulsions.

HYPOTHESIS: Hydrophobized fumed silica particles were previously reported for producing antibubbles that are quite stable in neutral as well as in acidic media. To produce acid-responsive antibubbles (e.g., for gastric drug delivery), the silica nanoparticles must be replaced by suitable particles, e.g., calcium carbonate (CaCO3), which can degrade at low pH to release the encapsulated drug. EXPERIMENTS: Two variants of CaCO3-stabilized antibubbles were prepared (by using CaCO3 particles pre-coated with stearic acid, or by using native CaCO3 particles in combination with sodium stearoyl lactylate) and drug release was compared with classic antibubbles produced with hydrophobized fumed silica particles. FINDINGS: CaCO3 particles (pre-coated with stearic acid) can be used to produce stable antibubbles, which provided an entrapment efficiency of a model drug (methylene blue, MB) of around 85%. A burst release of MB (∼60%) from the antibubbles was observed at pH 2 (i.e., the pH of the stomach), which was further increased to 80% during the next 30 min. On the contrary, at neutral pH, about 70% of the drug remained encapsulated for at least 2 h. We further demonstrated that the acidic conditions led to the desorption of CaCO3 particles from the air-liquid interface resulting in the destabilization of the antibubbles and the release of drug-containing cores.

Triple-Emulsion-Based Antibubbles: A Step Forward in Fabricating Novel Multi-Drug Delivery Systems.

Developing carriers capable of efficiently transporting both hydrophilic and lipophilic payloads is a captivating focus within the pharmaceutical and drug delivery research domain. Antibubbles, constituting an innovative encapsulation system designed for drug delivery purposes, have garnered scientific interest thanks to their distinctive water-in-air-in-water (W1/A/W2) structure. However, in contrast to their precursor, i.e., nanoparticle-stabilized W1/O/W2 double emulsion, traditional antibubbles lack the ability to accommodate a lipophilic payload, as the intermediary (volatile) oil layer of the emulsion is replaced by air during the antibubble fabrication process. Therefore, here, we report the fabrication of triple-emulsion-based antibubbles (O1/W1/A/W2), in which the inner aqueous phase was loaded with a nanoemulsion stabilized by various proteins, including whey, soy, or pea protein isolates. As model drugs, we employed the dyes Nile red in the oil phase and methylene blue in the aqueous phase. The produced antibubbles were characterized regarding their size distribution, entrapment efficiency, and stability. The produced antibubbles demonstrated substantial entrapment efficiencies for both lipophilic (ranging from 80% to 90%) and hydrophilic (ranging from 70% to 82%) components while also exhibiting an appreciable degree of stability during an extended rehydration period of two weeks. The observed variations among different antibubble variants were primarily attributed to differences in protein concentration rather than the type of protein used.

Low energy nebulization preserves integrity of SARS-CoV-2 mRNA vaccines for respiratory delivery.

Nebulization of mRNA therapeutics can be used to directly target the respiratory tract. A promising prospect is that mucosal administration of lipid nanoparticle (LNP)-based mRNA vaccines may lead to a more efficient protection against respiratory viruses. However, the nebulization process can rupture the LNP vehicles and degrade the mRNA molecules inside. Here we present a novel nebulization method able to preserve substantially the integrity of vaccines, as tested with two SARS-CoV-2 mRNA vaccines. We compare the new method with well-known nebulization methods used for medical respiratory applications. We find that a lower energy level in generating LNP droplets using the new nebulization method helps safeguard the integrity of the LNP and vaccine. By comparing nebulization techniques with different energy dissipation levels we find that LNPs and mRNAs can be kept largely intact if the energy dissipation remains below a threshold value, for LNP integrity 5-10 J/g and for mRNA integrity 10-20 J/g for both vaccines.

Advances in antibubble formation and potential applications.

Antibubbles are unusual physical objects consisting of a liquid core(s) surrounded by a thin air film/shell while in a bulk liquid. Antibubbles carry two air-liquid interfaces, i.e., one with the inner liquid and the other with the outer liquid. The distinct structure of antibubbles makes them quite attractive for drug and therapeutic delivery, although their potential applications have not been realized so far. The major challenge in this regard is a short-lived span of antibubbles, which is usually in the order of a few minutes to a few hours based on the stabilization mechanism used. We present a critical overview of different techniques that can be used to generate antibubbles. This includes a more commonly applied conventional approach in which the air-film is created through surface entrapment when a liquid jet/drop falls on a bulk liquid. The other available options rely on entirely different mechanisms for antibubble formation, for instance, through drop encapsulation by a submerged air bubble, or through evaporation/sublimation of volatile oil from a W/O/W double emulsion. Furthermore, the mechanisms of antibubble formation and collapse, and the factors affecting their stability have been discussed explicitly; and wherever required, the concept is correlated to other allied physical objects such as bubbles, liquid marbles, etc. Finally, the potential applications, research gaps in the existing knowledge, and some directions for future research are provided towards the end of this article.

Formulation and characterisation of drug-loaded antibubbles for image-guided and ultrasound-triggered drug delivery.

The aim of this study was to develop high load-capacity antibubbles that can be visualized using diagnostic ultrasound and the encapsulated drug can be released and delivered using clinically translatable ultrasound. The antibubbles were developed by optimising a silica nanoparticle stabilised double emulsion template. We produced an emulsion with a mean size diameter of 4.23 ± 1.63 µm where 38.9 ± 3.1% of the droplets contained a one or more cores. Following conversion to antibubbles, the mean size decreased to 2.96 ± 1.94 µm where 99% of antibubbles were <10 µm. The antibubbles had a peak attenuation of 4.8 dB/cm at 3.0 MHz at a concentration of 200 × 103 particles/mL and showed distinct attenuation spikes at frequencies between 5.5 and 13.5 MHz. No increase in subharmonic response was observed for the antibubbles in contrast to SonoVue®. High-speed imaging revealed that antibubbles can release their cores at MIs of 0.6. In vivo imaging indicated that the antibubbles have a long half-life of 68.49 s vs. 40.02 s for SonoVue®. The antibubbles could be visualised using diagnostic ultrasound and could be disrupted at MIs of ≥0.6. The in vitro drug delivery results showed that antibubbles can significantly improve drug delivery (p < 0.0001) and deliver the drug within the antibubbles. In conclusion antibubbles are a viable concept for ultrasound guided drug delivery.

Long-lived antibubbles: stable antibubbles through Pickering stabilization.

Antibubbles, which are liquid droplets surrounded by a thin shell of gas in a liquid phase, have several promising applications, among which is encapsulation. A major hurdle toward these applications has hitherto been the inherent instability of antibubbles, leading to lifetimes of at most minutes. Here we show the production of antibubbles with a lifetime of at least tens of hours, with their stability stemming from the adsorption of colloidal particles at gas-water interfaces. Antibubbles were produced by coating aqueous droplets with hydrophobic colloidal particles, gelling the droplets, and then dropping them into an aqueous colloidal suspension. This resulted in the formation of antibubbles with a long lifetime, also after the melting of the gel.

Experimental acoustic characterization of an endoskeletal antibubble contrast agent: First results.

PURPOSE: An antibubble is an encapsulated gas bubble with an incompressible inclusion inside the gas phase. Current-generation ultrasound contrast agents are bubble-based: they contain encapsulated gas bubbles with no inclusions. The objective of this work is to determine the linear and nonlinear responses of an antibubble contrast agent in comparison to two bubble-based ultrasound contrast agents, that is, reference bubbles and SonoVue TM . METHODS: Side scatter and attenuation of the three contrast agents were measured, using single-element ultrasound transducers, operating at 1.0, 2.25, and 3.5 MHz. The scatter measurements were performed at acoustic pressures of 200 and 300 kPa for 1.0 MHz, 300 kPa, and 450 kPa for 2.25 MHz, and 370 and 560 kPa for 3.5 MHz. Attenuation measurements were conducted at pressures of 13, 55, and 50 kPa for 1.0, 2.25, and 3.5 MHz, respectively. In addition, a dynamic contrast-enhanced ultrasound measurement was performed, imaging the contrast agent flow through a vascular phantom with a commercial diagnostic linear array probe. RESULTS: Antibubbles generated equivalent or stronger harmonic signal, compared to bubble-based ultrasound contrast agents. The second harmonic side-scatter amplitude of the antibubble agent was up to 3 dB greater than that of reference bubble agent and up to 4 dB greater than that of SonoVue TM at the estimated concentration of 8 × 10 4 bubbles/mL. For ultrasound with a center transmit frequency of 1.0 MHz, the attenuation coefficient of the antibubble agent was 8.7 dB/cm, whereas the attenuation coefficient of the reference agent was 7.7 and 0.3 dB/cm for SonoVue TM . At 2.25 MHz, the attenuation coefficients were 9.7, 3.0, and 0.6 dB/cm, respectively. For 3.5 MHz, they were 4.4, 1.8, and 1.0 dB/cm, respectively. A dynamic contrast-enhanced ultrasound recording showed the nonlinear signal of the antibubble agent to be 31% greater than for reference bubbles and 23% lower than SonoVue TM at a high concentration of 2 × 10 6 bubbles/mL. CONCLUSION: Endoskeletal antibubbles generate comparable or greater higher harmonics than reference bubbles and SonoVue TM . As a result, antibubbles with liquid therapeutic agents inside the gas phase have high potential to become a traceable therapeutic agent.

Microcapsules from self-assembled colloidal particles using aqueous phase-separated polymer solutions.

We report a new way of producing microcapsules consisting of a shell of aggregated colloids (also referred to as colloidosomes) using aqueous phase-separated polymer solutions (water-in-water emulsions) as a template. The extremely low interfacial tension of the template allows the production of reversible colloidosomes that spontaneously disintegrate depending on environmental conditions, making them ideal for controlled release purposes. Also, colloidosomes can have an elongated shape such that they may be used as microfluidic membranes or artificial arteries. Because the method described here does not use any organic solvents, it enables the use of sensitive materials such as cells and proteins.