Ambient air plasmas have been known to kill cancer cells. To enhance selectivity we have used antibody-conjugated nanoparticles. We achieved five times enhancement of melanoma cell death over the case of the plasma alone by using an air plasma with gold nanoparticles bound to anti-FAK antibodies. Our results show that this new interdisciplinary technique has enormous potential for use as a complement to conventional therapies.
Living cells can spatiotemporally control biochemical reactions to dynamically assemble membraneless organelles and remodel cytoskeleton. Herein, we present a microfluidic approach to prepare semi-permeable polymersomes comprising of amphiphilic triblock copolymer to achieve external signal-driven complex coacervation as well as biophysical reconstitution of cytoskeleton within the polymersomes. We also show that the microfluidic synthesis of polymersomes enables precise control over size, efficient encapsulation of enzymes as well as regulation of substrates without the use of biopores. Moreover, we demonstrate that the resulting triblock copolymer-based membrane in polymersomes is size-selective, allowing phosphoenol pyruvate to readily diffuse through the membrane and induce enzymatic reaction and successive coacervation or actin polymerization in the presence of pyruvate kinase and adenosine diphosphate inside the polymersomes. We envision that the Pluronic-based polymersomes presented in this work will shed light in the design of in vitro enzymatic reactions in artificial cell-like vesicles.
Recent advances in droplet microfluidics have led to the fabrication of versatile vesicles with a structure that mimics the cellular membrane. These artificial cell-like vesicles including polymersomes and liposomes effectively enclose an aqueous core with well-defined size and composition from the surrounding environment to implement various biological reactions, serving as a diverse functional reactor. The advantage of realizing various biological phenomena within a compartment separated by a membrane that resembles a natural cell membrane is actively explored in the fields of synthetic biology as well as biomedical applications including drug delivery, biosensors, and bioreactors, to name a few. In this Perspective, we first summarize various methods utilized in producing these polymersomes and liposomes. Moreover, we will highlight some of the recent advances in the design of these artificial cell-like vesicles for functional bioreactors and discuss the current issues and future perspectives.
Aqueous two-phase systems (ATPSs) have been widely used in the separation, purification, and enrichment of biomolecules for their excellent biocompatibility. While ultracentrifugation and microfluidic devices have been combined with ATPS to facilitate the separation of biomolecules and achieve high recovery yields, they often lack the ability to effectively isolate and separate biomolecules in low concentrations. In this work, we present a strategy that leverages the preferential partitioning of biomolecules in ATPS droplets to efficiently separate model extracellular vesicle (EV) particles. We demonstrate that the additional oil phase between the inner ATPS droplets and the aqueous continuous phase in triple emulsion droplets resolves the size controllability and instability issues of ATPS droplets, enabling the production of highly monodisperse ATPS-based polymersomes with enhanced stability for effective isolation of ATPS droplets from the surrounding environment. Furthermore, we achieve separation of model EV particles in a single dextran (DEX)-rich droplet by the massive production of ATPS-based polymersomes and osmotic-pressure-induced rupture of the selected polymersome in a hypertonic solution composed of poly(ethylene glycol) (PEG).
The recent development of RNA-based therapeutics in delivering nucleic acids for gene editing and regulating protein translation has led to the effective treatment of various diseases including cancer, inflammatory and genetic disorder, as well as infectious diseases. Among these, lipid nanoparticles (LNP) have emerged as a promising platform for RNA delivery and have shed light by resolving the inherent instability issues of naked RNA and thereby enhancing the therapeutic potency. These LNP consisting of ionizable lipid, helper lipid, cholesterol, and poly(ethylene glycol)-anchored lipid can stably enclose RNA and help them release into the cells' cytosol. Herein, the significant progress made in LNP research starting from the LNP constituents, formulation, and their diverse applications is summarized first. Moreover, the microfluidic methodologies which allow precise assembly of these newly developed constituents to achieve LNP with controllable composition and size, high encapsulation efficiency as well as scalable production are highlighted. Furthermore, a short discussion on current challenges as well as an outlook will be given on emerging approaches to resolving these issues.
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