Delivery vehicle and route of administration influences self-amplifying RNA biodistribution, expression kinetics, and reactogenicity.

Self-amplifying RNA (saRNA) is a next-generation RNA platform derived from an alphavirus that enables replication in host cytosol, offering a promising shift from traditional messenger RNA (mRNA) therapies by enabling sustained protein production from minimal dosages. The approval of saRNA-based vaccines, such as the ARCT-154 for COVID-19 in Japan, underscores its potential for diverse therapeutic applications, including vaccine development, cancer immunotherapy, and gene therapy. This study investigates the role of delivery vehicle and administration route on saRNA expression kinetics and reactogenicity. Employing ionizable lipid-based nanoparticles (LNPs) and polymeric nanoparticles, we administered saRNA encoding firefly luciferase to BALB/c mice through six routes (intramuscular (IM), intradermal (ID), intraperitoneal (IP), intranasal (IN), intravenous (IV), and subcutaneous (SC)), and observed persistent saRNA expression over a month. Our findings reveal that while LNPs enable broad route applicability and stability, pABOL (poly (cystamine bisacrylamide-co-4-amino-1-butanol)) formulations significantly amplify protein expression via intramuscular delivery. Notably, the disparity between RNA biodistribution and protein expression highlight the nuanced interplay between administration routes, delivery vehicles, and therapeutic outcomes. Additionally, our research unveiled distinct biodistribution profiles and inflammatory responses contingent upon the chosen delivery formulation and route. This research illuminates the intricate dynamics governing saRNA delivery, biodistribution and reactogenicity, offering essential insights for optimizing therapeutic strategies and advancing the clinical and commercial viability of saRNA technologies.

Clinical perspective on topical vaccination strategies.

Vaccination is one of the most successful measures in modern medicine to combat diseases, especially infectious diseases, and saves millions of lives every year. Vaccine design and development remains critical and involves many aspects, including the choice of platform, antigen, adjuvant, and route of administration. Topical vaccination, defined herein as the introduction of a vaccine to any of the three layers of the human skin, has attracted interest in recent years as an alternative vaccination approach to the conventional intramuscular administration because of its potential to be needle-free and induce a superior immune response against pathogens. In this review, we describe recent progress in developing topical vaccines, highlight progress in the development of delivery technologies for topical vaccines, discuss potential factors that might impact the topical vaccine efficacy, and provide an overview of the current clinical landscape of topical vaccines.

Quantification of surface composition and segregation on AuAg bimetallic nanoparticles by MALDI MS.

In this work we show that it is possible to use MALDI-TOF as a tool to quantify the atomic composition and to describe the phase segragation of the surface of ligand-coated, bimetallic AuAg nanoparticles. Our investigation shows that AuAg nanoparticles of various compositions exhibit core-shell heterogeneity with surface enrichment of Ag. A Monte-Carlo type simulation demonstrates that the surface Au and Ag atoms arrange in a random fashion.

Comparative characterisation of non-monodisperse gold nanoparticle populations by X-ray scattering and electron microscopy.

Accurate nanoparticle size determination is essential across various research domains, with many functionalities in nanoscience and biomedical research being size-dependent. Although electron microscopy is capable of resolving a single particle down to the sub-nm scale, the reliable representation of entire populations is plagued by challenges in providing statistical significance, suboptimal preparation procedures and operator bias. While alternative techniques exist that provide ensemble information in solution, their implementation is generally challenging for non-monodisperse populations. Herein, we explore the use of small-angle X-ray scattering in combination with form-free Monte Carlo fitting of scattering profiles as an alternative to conventional electron microscopy imaging in providing access to any type of core size distribution. We report on a cross-method comparison for quasi-monodisperse, polydisperse and bimodal gold nanoparticles of 2-7 nm in diameter and discuss advantages and limitations of both techniques.

Polymeric Micelles Loading Proteins through Concurrent Ion Complexation and pH-Cleavable Covalent Bonding for In Vivo Delivery.

Protein drugs have great potential as targeted therapies, yet their application suffers from several drawbacks, such as instability, short half-life, and adverse immune responses. Thus, protein delivery approaches based on stimuli-responsive nanocarriers can provide effective strategies for selectively enhancing the availability and activation of proteins in targeted tissues. Herein, polymeric micelles with the ability of encapsulating proteins are developed via concurrent ion complexation and pH-cleavable covalent bonding between proteins and block copolymers directed to pH-triggered release of the protein payload. Carboxydimethylmaleic anhydride (CDM) is selected as the pH-sensitive moiety, since the CDMamide bond is stable at physiological pH (pH 7.4), while it cleaves at pH 6.5, that is, the pathophysiological pH of tumors and inflammatory tissues. By using poly(ethylene glycol)-poly(l-lysine) block copolymers having 45% CDM addition, different proteins with various sizes and isoelectric points are loaded successfully. By using myoglobin-loaded micelles (myo/m) as a model, the stability of the micelles in physiological conditions and the dissociation and release of functional myoglobin at pH 6.5 are successfully confirmed. Moreover, myo/m shows extended half-life in blood compared to free myoglobin and micelles assembled solely by polyion complex, indicating the potential of this system for in vivo delivery of proteins.

On the effect of ligand shell heterogeneity on nanoparticle/protein binding thermodynamics.

Nonspecific protein adhesion to nanoparticle (NP) has been proven to have important implications in nanomedicine. However, there are only a few examples of careful studies relating protein binding thermodynamics to NP physicochemical features. In particular, a systematic investigation of how NP/protein binding parameters scale with size for sub-10 nm NPs and whether this scaling is affected by the surface feature of NPs remain unaddressed. Previously, we have developed an analytical ultracentrifugation (AUC) based method to determine NP/protein binding thermodynamic parameters that was shown to be particularly effective for sub-10 nm NPs. In this work, we exclusively utilize this method to investigate the binding parameters for a well-defined set of gold NPs with varying size and surface ligand ratios to the model protein human serum albumin. We find that gold NPs with a homogenous distribution of hydrophilic molecules in their ligand shell have a monotonic dependence of their binding constants and of the maximum number of bound proteins as a function of their surface area. On the other hand, a more complex relation is found for particles with patchy ligand shell. The findings of this research highlight the significance of surface morphology on the interplay between protein binding behavior and NP size.

Draw-Spinning of Kilometer-Long and Highly Stretchable Polymer Submicrometer Fibers.

A new method is developed to directly spinning perfectly uniaxial fibers in an ultrafast manner. Besides, this method can tune the fibers' diameter through adjusting processing parameters such as the feeding rate of precursors. Uniaxial nylon 66 fibers prepared via this method show superior mechanical properties due to the alignment in each level of the structure.

Continuous Draw Spinning of Extra-Long Silver Submicron Fibers with Micrometer Patterning Capability.

Ultrathin metal fibers can serve as highly conducting and flexible current and heat transport channels, which are essential for numerous applications ranging from flexible electronics to energy conversion. Although industrial production of metal fibers with diameters of down to 2 µm is feasible, continuous production of high-quality and low-cost nanoscale metal wires is still challenging. Herein, we report the continuous draw spinning of highly conductive silver submicron fibers with the minimum diameter of ∼200 nm and length of more than kilometers. We obtained individual AgNO3/polymer fibers by continuous drawing from an aqueous solution at a speed of up to 8 m/s. With subsequent heat treatment, freestanding Ag submicron fibers with high mechanical flexibility and electric conductivity have been obtained. Woven mats of aligned Ag submicron fibers were used as transparent electrodes with high flexibility and high performance with sheet resistance of 7 Ω sq-1 at a transparency of 96%. Continuous draw spinning opened new avenues for scalable, flexible, and ultralow-cost fabrication of extra-long conductive ultrathin metal fibers.

Highly Flexible Indium Tin Oxide Nanofiber Transparent Electrodes by Blow Spinning.

Transparent conductive film (TCF) has found wide applications. Indium tin oxide (ITO) is currently the most widely used transparent electrode. However, major problem of ITO is the lacking of flexibility, which totally limits its applications. Here, we report a highly flexible transparent electrode consisting of freestanding ITO nanofiber network fabricated by blow spinning, the advantage of which is its high-efficiency, low cost and safety. When the bending radius decreased to 0.5 mm, the resistance of the transparent electrodes only increased by 18.4%. Furthermore, the resistance was almost unchanged after thousands of bending cycles at 3.5 mm bending radius.

Large-Scale Spinning of Silver Nanofibers as Flexible and Reliable Conductors.

Conducting metal nanowires can be assembled into thin films for flexible electronics and optoelectronics applications including transparent electrodes, nanocircuits, and electronic skin, however, the junction resistances and low aspect ratios still limit its performance. Herein we report high-quality silver nanofibers (AgNFs) synthesized by a gas-assistant solution spinning method. Compared with traditional Ag nanowires that usually have lengths below 100 µm, AgNFs are infinitely long and can be easily assembled into large-scale 2D and 3D flexible conductors with fused junctions between nanofibers. The AgNF networks showed high transparency, low sheet resistance (e. g, 6 Ω sq(-1) at ∼97% transparency), and high flexibility as transparent electrodes, whereas the 3D AgNF sponge could be used as a deformable and robust 3D conductor.