Hydrogels for RNA delivery.

RNA-based therapeutics have shown tremendous promise in disease intervention at the genetic level, and some have been approved for clinical use, including the recent COVID-19 messenger RNA vaccines. The clinical success of RNA therapy is largely dependent on the use of chemical modification, ligand conjugation or non-viral nanoparticles to improve RNA stability and facilitate intracellular delivery. Unlike molecular-level or nanoscale approaches, macroscopic hydrogels are soft, water-swollen three-dimensional structures that possess remarkable features such as biodegradability, tunable physiochemical properties and injectability, and recently they have attracted enormous attention for use in RNA therapy. Specifically, hydrogels can be engineered to exert precise spatiotemporal control over the release of RNA therapeutics, potentially minimizing systemic toxicity and enhancing in vivo efficacy. This Review provides a comprehensive overview of hydrogel loading of RNAs and hydrogel design for controlled release, highlights their biomedical applications and offers our perspectives on the opportunities and challenges in this exciting field of RNA delivery.

Oxidative-Species-Selective Materials for Diagnostic and Therapeutic Applications.

With the increasing recognition of the diverse roles and significance of oxidative species in the pathogenesis of many diseases, a tremendous amount of work on the development of oxidative-species-responsive materials has been conducted for 1) detecting oxygen metabolites or diagnosis of oxidative-stress-relevant diseases, 2) reducing oxidative stress in the disease sites, and/or 3) delivering therapeutic and diagnostic agents. In this review, we first discuss the distinct features and biological functions of each oxidative species. Then the selectivity and sensitivity of chemical linkers/groups to specific oxidative species and the underlying chemistry of their particular interactions are systematically elucidated. Their potential biomedical applications are also highlighted. We expect that this comprehensive review will provide more insights for the design and development of oxidative-species-selective materials for more effective diagnostic and therapeutic applications.

One step synthesis of antimicrobial peptide protected silver nanoparticles: The core-shell mutual enhancement of antibacterial activity.

Over the past few decades, the overuse of antibiotics has led to the emergence of resistant bacteria and environmental issues. Both silver nanoparticles (AgNPs) and antimicrobial peptides (AMPs) hold potential to replace antibiotics. Combining both AMPs and AgNPs into a composite material may create novel properties such as enhanced antibacterial activity, lower cytotoxicity and favorable stability in aqueous solution. We designed a 13 amino acid peptide (in short, P-13) with two functional regions: one is for antibacterial activity, and the other for reducing and stabilizing AgNPs with containing cysteine (C) residues in its C-terminus. With a single step reaction, we have successfully synthesized P-13 protected AgNPs (P-13@AgNPs) with a hydrodynamic diameter of about 11 nm. In the preliminary antibacterial activity assay, the minimum inhibitory concentrations (MICs) of P-13@AgNPs were up to 7.8 µg/mL against E. coli, S. aureus and B. pumilus, and 15.6 µg/mL against P. aeruginosa. Moreover, Flow cytometry analysis of E. coli, S. aureus, P. aeruginosa and B. pumilus show that the mortality of the strains reached 96 %, 96 %, 91 % and 90 %, respectively. The cytotoxicity of AgNPs was reduced dramatically after protected by P-13, and P-13 was favorable for the stability of the AgNPs solution. We believe this work could set up an example to make the best use of the individual material's properties to produce novel nanocomposites with better antibacterial activity.

Generating Cyan Fluorescence with De Novo Tripeptides: An In Vitro Mutation Study on the Role of Single Amino Acid Residues and Their Sequence.

Amino acids are natural choices as building blocks when developing biofunctional entities owing to their superior diversity and versatile physicochemical properties compared to nucleotide bases. A simple permutation of the amino acids creates a broad palette of proteins and these have been successfully engineered into useful biofunctional agents. For example, the intrinsic ultraviolet fluorescence of phenylalanine and tryptophan has been engineered to emit in the visible spectrum, which has broad applications for imaging/sensing probes, photothermal therapy agents, optogenetic switches, etc. Nature produces more colorful coats/furs, feathers/hairs, and eyes through various biochemical modifications of tyrosine-based pigmentation. However, it is challenging to modulate the fluorescence wavelength from the UV to the visible region through oligopeptides. Herein, we report an innovative approach to obtain cyan fluorescence by using de novo tripeptides containing glycine, tyrosine, and lysine, which form robust dimer structures under moderate oxidizing conditions. Through an in vitro mutation approach, we deduce that both the amino acids and their sequence play significant roles in modulating the fluorescence. We believe this work holds great promise for developing novel cell imaging and resonance energy-transfer-based fluorescent probes.

A novel ultrasensitive surface plasmon resonance-based nanosensor for nitrite detection.

Nitrite is a common food additive, however, its reduction product, nitrosamine, is a strong carcinogen, and hence the ultra-sensitive detection of nitrite is an effective means to prevent related cancers. In this study, different sized gold nanoparticles (AuNPs) were modified with P-aminothiophenol (ATP) and naphthylethylenediamine (NED). In the presence of nitrite, satellite-like AuNPs aggregates formed via the diazotization coupling reaction and the color of the system was changed by the functionalized AuNPs aggregates. The carcinogenic nitrite content could be detected by colorimetry according to the change in the system color. The linear concentration range of sodium nitrite was 0-1.0 µg mL-1 and the detection limit was determined to be 3.0 ng mL-1. Compared with the traditional method, this method has the advantages of high sensitivity, low detection limit, good selectivity and can significantly lower the naked-eye detection limit to 3.0 ng mL-1. In addition, this method is suitable for the determination of nitrite in various foods. We think this novel designed highly sensitive nitrate nanosensor holds great market potential.

Self-Assembly of Enzyme-Like Nanofibrous G-Molecular Hydrogel for Printed Flexible Electrochemical Sensors.

Conducting hydrogels provide great potential for creating designer shape-morphing architectures for biomedical applications owing to their unique solid-liquid interface and ease of processability. Here, a novel nanofibrous hydrogel with significant enzyme-like activity that can be used as "ink" to print flexible electrochemical devices is developed. The nanofibrous hydrogel is self-assembled from guanosine (G) and KB(OH)4 with simultaneous incorporation of hemin into the G-quartet scaffold, giving rise to significant enzyme-like activity. The rapid switching between the sol and gel states responsive to shear stress enables free-form fabrication of different patterns. Furthermore, the replication of the G-quartet wires into a conductive matrix by in situ catalytic deposition of polyaniline on nanofibers is demonstrated, which can be directly printed into a flexible electrochemical electrode. By loading glucose oxidase into this novel hydrogel, a flexible glucose biosensor is developed. This study sheds new light on developing artificial enzymes with new functionalities and on fabrication of flexible bioelectronics.

Logic Catalytic Interconversion of G-Molecular Hydrogel.

By incorporating hemin into G-quadruplex (G4) during cation-templated self-assembly between guanosine and KB(OH)4, we have constructed an artificial enzyme hydrogel (AEH)-based system for the highly sensitive and selective detection of Pb2+. The sensing strategy is based on a Pb2+-induced decrease in AEH activity. Because of the higher efficiency of Pb2+ for stabilizing G4 compared with K+, the Pb2+ ions substitute K+ and trigger hemin release from G4, thus giving rise to a conformational interconversion accompanied by the loss of enzyme activity. The Pb2+-induced catalytic interconversion endows the AEH-based system with high sensitivity and selectivity for detecting Pb2+. As a result, the AEH-based system shows an excellent response for Pb2+ in the range from 1 pM to 50 nM with a limit of detection of ∼0.32 pM, which is much lower than that of the previously reported G4-DNAzyme. We also demonstrate that this AEH-based system exhibits high selectivity toward Pb2+ over other metal ions. Furthermore, two two-input INHIBIT logic gates have been constructed via switching of the catalytic interconversion induced by K+ and Pb2+ or K+ and pH. Given its versatility, this AEH-based system provides a novel platform for sensing and biomolecular computation.

Fluorescence resonance energy transfer between bovine serum albumin and fluoresceinamine.

Physical binding-mediated organic dye direct-labelling of proteins could be a promising technology for bio-nanomedical applications. Upon binding, it was found that fluorescence resonance energy transfer (FRET) occurred between donor bovine serum albumin (BSA; an amphiphilic protein) and acceptor fluoresceinamine (FA; a hydrophobic fluorophore), which could explain fluorescence quenching found for BSA. FRET efficiency and the distance between FA and BSA tryptophan residues were determined to 17% and 2.29 nm, respectively. Using a spectroscopic superimposition method, the saturated number of FAs that bound to BSA was determined as eight to give a complex formula of FA8-BSA. Finally, molecular docking between BSA and FA was conducted, and conformational change that occurred in BSA upon binding to FA molecules was also studied by three-dimensional fluorescence microscopy.

Understanding the Robust Physisorption between Bovine Serum Albumin and Amphiphilic Polymer Coated Nanoparticles.

The robust physisorption between nanoparticles (NPs) and proteins has attracted increasing attention due to the significance for both conjugation techniques and protein's corona formation at the bionano interface. In the present study, we first explored the possible binding sites of the bovine serum albumin (BSA) on amphiphilic polymer coated gold nanoparticles (AP-AuNPs). By using mass spectrometry, a 105-amino-acid peptide (12.2 kDa) is discovered as the possible "epitope" responsible for the robust physisorption between BSA and AP-AuNPs. Second, with the help of nanometal surface energy transfer (NSET) theory, we further found that the epitope peptide could insert at least 2.9 nm into the organic molecular layers of AP-AuNPs when the robust conjugates formed, which indicates how such a long epitope peptide can be accommodated by AP-AuNPs and resist protease's digestion. These findings might shed light on a new strategy for studying interactions between proteins and NPs, and further guide the rational design of NPs for safe and effective biomedical applications.

A fluorimetric study on the interaction between a Trp-containing beta-strand peptide and amphiphilic polymer-coated gold nanoparticles.

Owing to the inevitability of nanoparticles encountering proteins/peptides in current bio-nano-medicine development, it is important to know how they interact with each other in vitro before developing in vivo applications. To this end, a model de novo ß-sheet-forming peptide and typical biocompatible nanoparticles were selected to study thermodynamic aspects of their interactions via a fluorescence quenching method. The results showed that Pep11 and AuNPs spontaneously formed conjugates, mainly driven by a coulombic interaction with a binding affinity of ~ 0.1 µM(-1); the physical adsorption process was cooperative. These results deepen our quantitative understanding of nanoparticle-peptide interactions. The results may also be helpful in further nanoparticle-peptide hybrid nanofabrication and also useful for the application of nanoparticles in the treatment of amyloid diseases.

Discrete nanoparticle-BSA conjugates manipulated by hydrophobic interaction.

Nanoparticle-protein conjugates are promising probes for biological diagnostics as well as versatile building blocks for nanotechnology. Here we demonstrate a facile method to prepare nanoparticles bearing discrete numbers of BSA simply by physical adsorption and electrophoretic isolation, in which the specific amphiphilic properties of BSA play important roles and the number of adsorbed BSA molecules can also be manipulated by tuning the coating extent of nanoparticles by amphiphilic polymer.