Nano-Assembled Polyphosphazene Delivery System Enables Effective Intranasal Immunization with Nipah Virus Subunit Vaccine.

The ultimate vaccine against infections caused by Nipah virus should be capable of providing protection at the respiratory tract─the most probable port of entry for this pathogen. Intranasally delivered vaccines, which target nasal-associated lymphoid tissue and induce both systemic and mucosal immunity, are attractive candidates for enabling effective vaccination against this lethal disease. Herein, the water-soluble polyphosphazene delivery vehicle assembles into nanoscale supramolecular constructs with the soluble extracellular portion of the Hendra virus attachment glycoprotein─a promising subunit vaccine antigen against both Nipah and Hendra viruses. These supramolecular constructs signal through Toll-like receptor 7/8 and promote binding interactions with mucin─an important feature of effective mucosal adjuvants. High mass contrast of phosphorus-nitrogen backbone of the polymer enables a successful visualization of nanoconstructs in their vitrified state by cryogenic electron microscopy. Here, we characterize the self-assembly of polyphosphazene macromolecule with biologically relevant ligands by asymmetric flow field flow fractionation, dynamic light scattering, fluorescence spectrophotometry, and turbidimetric titration methods. Furthermore, a polyphosphazene-enabled intranasal Nipah vaccine candidate demonstrates the ability to induce immune responses in hamsters and shows superiority in inducing total IgG and neutralizing antibodies when benchmarked against the respective clinical stage alum adjuvanted vaccine. The results highlight the potential of polyphosphazene-enabled nanoassemblies in the development of intranasal vaccines.

Directly visualizing individual polyorganophosphazenes and their single-chain complexes with proteins.

Polyorganophosphazenes are water-soluble macromolecules with immunoadjuvant activity that self-assemble with proteins to enable biological functionality. Direct imaging by cryogenic electron microscopy uncovers the coil structure of those highly charged macromolecules. The successful visualization of individual polymer chains within the vitrified state is achieved in the absence of additives for contrast enhancement and is attributed to the high mass contrast of the inorganic backbone. Upon assembly with proteins, multiple protein copies bind at the single polymer chain level resulting in structures reminiscent of compact spherical complexes or stiffened coils. The outcome depends on protein characteristics and cannot be deduced by commonly used characterization techniques, such as light scattering, thus revealing direct morphological insights crucial for understanding biological activity. Atomic force microscopy supports the morphology outcomes while advanced analytical techniques confirm protein-polymer binding. The chain visualization methodology provides tools for gaining insights into the processes of supramolecular assembly and mechanistic aspects of polymer enabled vaccine delivery.

Engineering Degradation Rate of Polyphosphazene-Based Layer-by-Layer Polymer Coatings.

Degradable layer-by-layer (LbL) polymeric coatings have distinct advantages over traditional biomedical coatings due to their precision of assembly, versatile inclusion of bioactive molecules, and conformality to the complex architectures of implantable devices. However, controlling the degradation rate while achieving biocompatibility has remained a challenge. This work employs polyphosphazenes as promising candidates for film assembly due to their inherent biocompatibility, tunability of chemical composition, and the buffering capability of degradation products. The degradation of pyrrolidone-functionalized polyphosphazenes was monitored in solution, complexes and LbL coatings (with tannic acid), providing the first to our knowledge comparison of solution-state degradation to solid-state LbL degradation. In all cases, the rate of degradation accelerated in acidic conditions. Importantly, the tunability of the degradation rate of polyphosphazene-based LbL films was achieved by varying film assembly conditions. Specifically, by slightly increasing the ionization of tannic acid (near neutral pH), we introduce electrostatic "defects" to the hydrogen-bonded pairs that accelerate film degradation. Finally, we show that replacing the pyrrolidone side group with a carboxylic acid moiety greatly reduces the degradation rate of the LbL coatings. In practical applications, these coatings have the versatility to serve as biocompatible platforms for various biomedical applications and controlled release systems.

Cryo-EM and AFM visualize linear polyorganophosphazene: individual chains and single-chain assemblies with proteins.

Polyorganophosphazenes are biodegradable macromolecules with potent immunoadjuvant activity that self-assemble with protein antigens to provide biological activity. Direct imaging by cryogenic electron microscopy reveals the coil structure of the highly-charged high molecular mass synthetic polyorganophosphazenes within the vitrified state without any additives for contrast enhancement for the first time. Upon mixing with protein antigens under a controlled stoichiometric ratio, multiple proteins bind at the single chain level revealing a structural change reminiscent of compact spherical complexes or stiffened coils depending on the bound protein antigen. The structural outcome depends on the protein charge density that cannot be deduced by methods, such as dynamic light scattering, thus revealing direct morphological insight necessary to understand in vivo biological activity. Complementary atomic force microscopy supports the binding morphology outcomes as well as additional analytical techniques that indicate binding. These observations open opportunities to understand supramolecular assembly of proteins and other biomacromolecules at the single chain level with highly charged polyelectrolytes for vaccines as well as important to developing fields such as polyelectrolyte complex coacervation.

Integrating Antioxidant Functionality into Polymer Materials: Fundamentals, Strategies, and Applications.

While antioxidants are widely known as natural components of healthy food and drinks or as additives to commercial polymer materials to prevent their degradation, recent years have seen increasing interest in enhancing the antioxidant functionality of newly developed polymer materials and coatings. This paper provides a critical overview and comparative analysis of multiple ways of integrating antioxidants within diverse polymer materials, including bulk films, electrospun fibers, and self-assembled coatings. Polyphenolic antioxidant moieties with varied molecular architecture are in the focus of this Review, because of their abundance, nontoxic nature, and potent antioxidant activity. Polymer materials with integrated polyphenolic functionality offer opportunities and challenges that span from the fundamentals to their applications. In addition to the traditional blending of antioxidants with polymer materials, developments in surface grafting and assembly via noncovalent interaction for controlling localization versus migration of antioxidant molecules are discussed. The versatile chemistry of polyphenolic antioxidants offers numerous possibilities for programmed inclusion of these molecules in polymer materials using not only van der Waals interactions or covalent tethering to polymers, but also via their hydrogen-bonding assembly with neutral molecules. An understanding and rational use of interactions of polyphenol moieties with surrounding molecules can enable precise control of concentration and retention versus delivery rate of antioxidants in polymer materials that are critical in food packaging, biomedical, and environmental applications.

Ocean Salinity Sensing Using Long-Period Fiber Gratings Functionalized with Layer-by-Layer Hydrogels.

Rapid, accurate, and real-time measurements of ocean salinity are of great importance for a host of scientific, commercial, and defense applications. We demonstrate a highly sensitive, fast-responding fiber-optic salinity sensor that integrates long-period fiber gratings (LPFGs) with ionic strength-responsive hydrogel. The submicron-thick hydrogel was synthesized via layer-by-layer electrostatic assembly of partially quaternized poly(4-vinylpyridine) (qP4VP) and poly(acrylic acid), followed by chemical cross-linking. Spectroscopic ellipsometry measurement of a hydrogel made of 37% quaternized qP4VP showed robust and reversible swelling/deswelling in solutions with salt concentrations ranging from 0.4 to 0.8 M (22.8-44.7 g/kg) around pH 8.1. The swelling/deswelling process induced large changes in the refractive index of the hydrogel, leading to resultant shift in the resonance wavelength (RW) of LPFGs. The salinity-dependent optical response of the hydrogel-coated LPFGs is in good agreement with ellipsometry measurement. LPFGs coated with the hydrogel exhibited a sensitivity of 7 nm RW shift/M (125.5 pm/‰) with a measurement time less than 5 s. The shift in the resonance wavelength correlated linearly with salt concentration, making quantification of measured salinity straightforward.