Full-cycle study on developing a novel structured micromixer and evaluating the nanoparticle products as mRNA delivery carriers.

Achieving precise control of nanoparticle size while maintaining consistency and high uniformity is of paramount importance for improving the efficacy of nanoparticle-based therapies and minimizing potential side effects. Although microfluidic technologies are widely used for reliable nanoparticle synthesis, they face challenges in meeting critical homogeneity requirements, mainly due to imperfect mixing efficiency. Furthermore, channel clogging during continuous operation presents a significant obstacle in terms of quality control, as it progressively impedes the mixing behavior necessary for consistent nanoparticle production for therapeutic delivery and complicates the scaling-up process. This study entailed the development of a 3D-printed novel micromixer embedded with hemispherical baffle microstructures, a dual vortex mixer (DVM), which integrates Dean vortices to generate two symmetrical counter-rotating intensified secondary flows. The DVM with a relatively large mixer volume showed rapid mixing characteristics even at a flow rate of several mL min-1 and produced highly uniform lipids, liposomes, and polymer nanoparticles in a size range (50-130 nm) and polydispersity index (PDI) values below 0.15. For the evaluation of products, SARS-CoV-2 Spike mRNA-loaded lipid nanoparticles were examined to verify protein expression in vitro and in vivo using firefly luciferase (FLuc) mRNA. This showed that the performance of the system is comparable to that of a commercial toroidal mixer. Moreover, the vigorous in-situ dispersion of nanoparticles by harnessing the power of vortex physically minimizes the occurrence of aggregation, ensuring consistent production performance without internal clogging of a half-day operation and facilitating quality control of the nanoparticles at desired scales.

Magnetic delivery and ultrasound-responsive release of chelating microcapsules for selective removal of urolithiasis.

A novel urolithiasis treatment in which a chelating solution encapsulated in poly(lactic-co-glycolic acid); PLGA-based microcapsules was delivered magnetically to specific urolithiasis sites and then subjected to ultrasound (US) to release the chelating solution and dissolve the stones. Using a double-droplet microfluidics method, a hexametaphosphate (HMP) chelating solution was encapsulated in an Fe3O4 nanoparticle (Fe3O4 NP)-loaded PLGA polymer shell with a thickness of <15 µm, forming homogenous microcapsules of 319 ± 14 µm in size. The obtained microcapsules (HMP/Fe3O4@PLGA) exhibited efficient magnetic mobility and US-responsive solution release. Moreover, in a Ψ-shaped flow chip, selective delivery of HMP from the microcapsules was achieved with high magnetic delivery efficiency (>90%), and an effective removal efficacy (>95%, 7 repeat cycles) of artificial calcium oxalate (5 mm in size) via a chelating effect. Eventually, the potential removal of urolithiasis in the body was verified using a PDMS-based kidney urinary flow-imitating chip with a human kidney stone (CaOx 100%, 5-7 mm in size) located in the minor calyx under an artificial urine counter flow (0.5 mL min-1). In the end, more than 50% of the stone, even in surgically tricky regions, was removed by 10 repeated treatments. Therefore, the selective approach of stone-dissolution capsules will help to develop alternative urolithiasis treatments to conventional surgical and systemic dissolution approaches.

Laminar flow-assisted synthesis of amorphous ZIF-8-based nano-motor with enhanced transmigration for photothermal cancer therapy.

Because of their biocompatibility, there are promising applications in various fields for enzyme-powered nano-motors. However, enzymes can undergo denaturation under harsh conditions. Here, we report the flow-assisted synthesis of an enzyme-based amorphous ZIF-8 nano-motor (A-motor; Pdop@urease@aZIF-8) for enhanced movement and protection of polydopamine and enzymes. Multiple laminar flow types with varied input ratios effectively entrapped enzymes into amorphous ZIF-8 shells in a serial flow with a momentary difference. The obtained A-motor exhibited superior enzymatic activity and photothermal ablation properties with excellent durability due to the protection the amorphous shell offers from the external environment. Furthermore, in the bio-mimic 2D membrane model, the enhanced mobility of the A-motor afforded high transmigration (>80%), which had a powerful effect on bladder cancer cell ablation via photothermal therapy. This work envisages that the rapid flow approach will facilitate scalable manufacturing of the nano-motors under low stress to vulnerable biomolecules, which would be extended to nano-biomedical applications in various body environments.

An Active Absorbent for Cleanup of High-Concentration Strong Acid and Base Solutions.

There is significant interest in developing novel absorbents for hazardous material cleanup. Iron oxide-coated melamine formaldehyde sponge (MFS/IO) absorbents with various IO layer thicknesses were synthesized. Various other absorbents were also synthesized and compared to evaluate the absorption capability of the MFS/IO absorbents for strong acid (15%, v/v) and base (50%, m/m) solutions. Specifically, absorbent and solution drop tests, dust tests, and droplet fragment tests were performed. Among the various absorbents, MFS/IO absorbents possessing a needlelike surface morphology showed several unique characteristics not observed in other absorbents. The MFS/IO absorbents naturally absorbed a strong base solution (absorption time: 0.71-0.5 s, absorption capacity: 10,000-34,000%) without an additional external force and immediately absorbed a strong acid solution (0.31-0.43 s, 9830-10,810%) without absorption delay/overflow during absorbent and solution drop tests, respectively. The MFS/IO absorbents were also demonstrated to be ideal absorbents that generated fewer dust particles (semiclass 1 (ISO 3) level of 280 piece/L) than the level of a clean room (class 100). Furthermore, the MFS/IO absorbents were able to prevent the formation of droplet fragments and solution overflow during the solution drop test due to their unique surface morphology and extremely high absorption speed/capacity, respectively.

Polyelectrolyte Brush-Grafted Polydopamine-Based Catalysts with Enhanced Catalytic Activity and Stability.

Three types of surface treatments, namely, polyethyleneimine (PEI) coating, short PEI (S-PEI) grafting, and long PEI (L-PEI) grafting, were performed on polydopamine (Pdop)-based catalysts to enhance their catalytic activity and stability. Brush-grafted catalysts were prepared by the stepwise synthesis of Au and short (or long) PEI brushes on Pdop particles (PdopP/Au/S- or L-PEI grafting). PEI-coated Pdop-based catalysts (PdopP/Au/PEI coating) were also prepared as non-brush-grafted catalysts. Among the surface-treated PdopP/Au catalysts, the brush-grafted catalysts (S-PEI and L-PEI grafting) exhibited excellent and stable catalytic performance because the brush grafting enabled the protection of the catalysts against harsh conditions, effective transfer of reactants to the catalysts, and confinement of reactants around the catalysts. The brush-grafted catalysts could also more effectively decompose larger dyes than the non-brush-grafted catalysts. The process-to-effectiveness of PEI coating is the best because the release of Pdop from PdopP/Au was moderately inhibited by the presence of only one layer of PEI coating on the PdopP/Au. Thus, this approach could be an alternative method to enhance the stability of PdopP/Au catalysts.