in-situ-photo-rheology-monitors-viscoelastic-changes-photo-responsive

<em>In situ</em> Photo-rheology Monitors Viscoelastic Changes in Photo-responsive Polymer Networks

In situ photo-rheology is a powerful technique for characterizing the evolution of photo-responsive materials during irradiation. This technique enables ...

Stanford University

Natural selective filtering systems (e.g., the extracellular matrix, nuclear pores, and mucus) separate molecules selectively and efficiently, and the detailed understanding of transport mechanisms exploited in these systems provides important bioinspired design principles for selective filters. In particular, nucleoporins consist of consensus repeat sequences that are readily utilized for engineering repeat proteins. Here, the consensus repeat sequence of Nsp1, a yeast nucleoporin, is polymerized to form a nucleoporin-like protein (NLP) and mutated to understand the effect of sequence on selective transport. The hydrophilic spacers of the NLPs were redesigned considering net charge, charge distribution, and polarity. Mutations were made near to and far from the FSFG interacting domain to explore the role of highly conserved residues as a function of spatial proximity. A nuclear transport receptor-cargo complex, nuclear transport factor 2-green fluorescent protein (NTF2-GFP), was used as a model for changes in transport. For mutations of the charged spacer, some mutations of highly conserved charged residues were possible without knocking out selective transport of the NTF2, but the formation of regions of clustered negative charge has an unfavorable effect on nuclear transporter permeation. Thus, positive net charge and alternating positive and negative charge within the hydrophilic spacer are advantageous for recognition and selective transport. In the polarity panel, mutations that increased the interaction between NTF2-GFP and the gel led to decreased permeation of the NTF2-GFP due to blocking of the interface and inability of the NTF2-GFP to transport into the gel. Therefore, these results provide a strategy for tuning selective permeability of biomolecules using the artificially designed consensus repeat-based hydrogels.

Tuning Selective Transport of Biomolecules through Site-Mutated Nucleoporin-like Protein (NLP) Hydrogels.

Single polymer studies have revealed unexpected and heterogeneous dynamics among identical or seemingly similar macromolecules. In recent years, direct observation of single polymers has uncovered broad distributions in molecular behavior that play a key role in determining bulk properties. Early single polymer experiments focused primarily on biological macromolecules such as DNA, but recent advances in synthesis, imaging, and force spectroscopy have enabled broad exploration of chemically diverse polymer systems. In this Viewpoint, we discuss the recent study of synthetic polymers using single-molecule methods. In terms of polymer synthesis, direct observation of single chain polymerization has revealed heterogeneity in monomer insertion events at catalytic centers and decoupling of local and global growth kinetics. In terms of single polymer visualization, recent advances in super-resolution imaging, atomic force microscopy (AFM), and liquid-cell transmission electron microscopy (LC-TEM) can resolve structure and dynamics in single synthetic chains. Moreover, single synthetic polymers can be probed in the context of bulk material environments, including hydrogels, nanostructured polymers, and crystalline polymers. In each area, we highlight key challenges and exciting opportunities in using single polymer techniques to enhance our understanding of polymer science. Overall, the expanding versatility of single polymer methods will enable the molecular-scale design and fundamental understanding of a broad range of chemically diverse and functional polymeric materials.

100th Anniversary of Macromolecular Science Viewpoint: Single-Molecule Studies of Synthetic Polymers.

Embedding nanomaterials into polymer hydrogels enables the design of functional materials with tailored chemical, mechanical, and optical properties. Nanocapsules that protect interior cargo and disperse readily through a polymeric matrix have drawn particular interest for their ability to integrate chemically incompatible systems and to further expand the parameter space for polymer nanocomposite hydrogels. The properties of polymer nanocomposite hydrogels depend on the material composition and processing route, which were explored systematically in this work. The gelation kinetics of network-forming polymer solutions with and without silica-coated nanocapsules bearing polyethylene glycol (PEG) surface ligands were investigated using  dynamic rheology measurements. Network-forming polymers comprised either 4-arm or 8-arm star PEG with terminal anthracene groups, which dimerize upon irradiation with ultraviolet (UV) light. The PEG-anthracene solutions exhibited rapid gel formation upon UV exposure (365 nm); gel formation was observed as a crossover from liquid-like to solid-like behavior during  small-amplitude oscillatory shear rheology. This crossover time was non-monotonic with polymer concentration. Far below the overlap concentration (/* ≪ 1), spatially separated PEG-anthracene molecules were subject to forming intramolecular loops over intermolecular cross-links, thereby slowing the gelation process. Near the polymer overlap concentration (/* ∼ 1), rapid gelation was attributed to the ideal proximity of anthracene end groups from neighboring polymer molecules. Above the overlap concentration (/* > 1), increased solution viscosities hindered molecular diffusion, thereby reducing the frequency of dimerization reactions. Adding nanocapsules to PEG-anthracene solutions resulted in faster gelation than nanocapsule-free PEG-anthracene solutions with equivalent effective polymer concentrations. The final elastic modulus of nanocomposite hydrogels increased with nanocapsule volume fraction, signifying synergistic mechanical reinforcement by nanocapsules despite not being cross-linked into the polymer network. Overall, these findings quantify the impact of nanocapsule addition on the gelation kinetics and mechanical properties of polymer nanocomposite hydrogels, which are promising materials for applications in optoelectronics, biotechnology, and additive manufacturing.

Gelation Dynamics during Photo-Cross-Linking of Polymer Nanocomposite Hydrogels.

Polymeric protagonists for biological processes.

UV light can trigger a plethora of useful photochemical reactions for diverse applications, including photocatalysis, photopolymerization, and drug delivery. These applications typically require penetration of high-energy photons deep into materials, yet delivering these photons beyond the surface is extremely challenging due to absorption and scattering effects. Triplet-triplet annihilation upconversion (TTA-UC) shows great promise to circumvent this issue by generating high-energy photons from incident lower-energy photons. However, molecules that facilitate TTA-UC usually have poor water solubility, limiting their deployment in aqueous environments. To address this challenge, a nanoencapsulation method is leveraged to fabricate water-compatible UC micelles, enabling on-demand UV photon generation deep into materials. Two iridium-based complexes are presented for use as TTA-UC sensitizers with increased solubilities that facilitate the formation of highly emissive UV-upconverting micelles. Furthermore, this encapsulation method is shown to be generalizable to nineteen UV-emitting UC systems, accessing a range of upconverted UV emission profiles with wavelengths as low as 350 nm. As a proof-of-principle demonstration of precision photochemistry at depth, UV-emitting UC micelles are used to photolyze a fluorophore at a focal point nearly a centimeter beyond the surface, revealing opportunities for spatially controlled manipulation deep into UV-responsive materials.

Spatially Controlled UV Light Generation at Depth using Upconversion Micelles.

Elastin-like polypeptides (ELPs) are a promising material platform for engineering stimuli-responsive biomaterials, as ELPs undergo phase separation above a tunable transition temperature. ELPs with phase behavior that is isothermally regulated by biological stimuli remain attractive for applications in biological systems. Herein, we report protease-driven phase separation of ELPs. Protease-responsive "cleavable" ELPs comprise a hydrophobic ELP block connected to a hydrophilic ELP block by a protease cleavage site linker. The hydrophilic ELP block acts as a solubility tag for the hydrophobic ELP block, creating a temperature window in which the cleavable ELP reactant is soluble and the proteolytically generated hydrophobic ELP block is insoluble. Within this temperature window, isothermal, protease-driven phase separation occurs when a critical concentration of hydrophobic cleavage product accumulates. Furthermore, protease-driven phase separation is generalizable to four compatible protease-cleavable ELP pairings. This work presents exciting opportunities to regulate ELP phase behavior in biological systems using proteases.

Protease-Driven Phase Separation of Elastin-Like Polypeptides.

Biomolecules present promising stimuli-responsive mechanisms to revolutionize soft actuators. Proteins, peptides, and nucleic acids foster specific intermolecular interactions, and their boundless sequence design spaces encode precise actuation capabilities. Drawing inspiration from nature, biomolecular actuators harness existing stimuli-responsive properties to meet the needs of diverse applications. This review features biomolecular actuators that respond to a wide variety of stimuli to drive both user-directed and autonomous actuation. We discuss how advances in biomaterial fabrication accelerate prototyping of precise, custom actuators, and we identify biomolecules with untapped actuation potential. Finally, we highlight opportunities for multifunctional and reconfigurable biomolecules to improve the versatility and sustainability of next-generation soft actuators.

Article	Year
Tuning Selective Transport of Biomolecules through Site-Mutated Nucleoporin-like Protein (NLP) Hydrogels. Biomacromolecules\| PubMed ID: 33428378	2021
100th Anniversary of Macromolecular Science Viewpoint: Single-Molecule Studies of Synthetic Polymers. ACS macro letters\| PubMed ID: 35638639	2020
Gelation Dynamics during Photo-Cross-Linking of Polymer Nanocomposite Hydrogels. ACS polymers Au\| PubMed ID: 37065714	2023
Polymeric protagonists for biological processes. Nature chemistry\| PubMed ID: 37248342	2023
Spatially Controlled UV Light Generation at Depth using Upconversion Micelles. Advanced materials (Deerfield Beach, Fla.)\| PubMed ID: 37548335	2023
Protease-Driven Phase Separation of Elastin-Like Polypeptides. Biomacromolecules\| PubMed ID: 38980747	2024
Biomolecular Actuators for Soft Robots. Chemical reviews\| PubMed ID: 40331746	2025

Danielle J. Mai

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