Mechanical release of homogenous proteins from supramolecular gels.

A long-standing challenge is how to formulate proteins and vaccines to retain function during storage and transport and to remove the burdens of cold-chain management. Any solution must be practical to use, with the protein being released or applied using clinically relevant triggers. Advanced biologic therapies are distributed cold, using substantial energy, limiting equitable distribution in low-resource countries and placing responsibility on the user for correct storage and handling. Cold-chain management is the best solution at present for protein transport but requires substantial infrastructure and energy. For example, in research laboratories, a single freezer at -80 °C consumes as much energy per day as a small household1. Of biological (protein or cell) therapies and all vaccines, 75% require cold-chain management; the cost of cold-chain management in clinical trials has increased by about 20% since 2015, reflecting this complexity. Bespoke formulations and excipients are now required, with trehalose2, sucrose or polymers3 widely used, which stabilize proteins by replacing surface water molecules and thereby make denaturation thermodynamically less likely; this has enabled both freeze-dried proteins and frozen proteins. For example, the human papilloma virus vaccine requires aluminium salt adjuvants to function, but these render it unstable against freeze-thaw4, leading to a very complex and expensive supply chain. Other ideas involve ensilication5 and chemical modification of proteins6. In short, protein stabilization is a challenge with no universal solution7,8. Here we designed a stiff hydrogel that stabilizes proteins against thermal denaturation even at 50 °C, and that can, unlike present technologies, deliver pure, excipient-free protein by mechanically releasing it from a syringe. Macromolecules can be loaded at up to 10 wt% without affecting the mechanism of release. This unique stabilization and excipient-free release synergy offers a practical, scalable and versatile solution to enable the low-cost, cold-chain-free and equitable delivery of therapies worldwide.

Chemical control of structure and guest uptake by a conformationally mobile porous material.

Metal-organic frameworks (MOFs) are crystalline synthetic porous materials formed by binding organic linkers to metal nodes: they can be either rigid1,2 or flexible3. Zeolites and rigid MOFs have widespread applications in sorption, separation and catalysis that arise from their ability to control the arrangement and chemistry of guest molecules in their pores via the shape and functionality of their internal surface, defined by their chemistry and structure4,5. Their structures correspond to an energy landscape with a single, albeit highly functional, energy minimum. By contrast, proteins function by navigating between multiple metastable structures using bond rotations of the polypeptide6,7, where each structure lies in one of the minima of a conformational energy landscape and can be selected according to the chemistry of the molecules that interact with the protein. These structural changes are realized through the mechanisms of conformational selection (where a higher-energy minimum characteristic of the protein is stabilized by small-molecule binding) and induced fit (where a small molecule imposes a structure on the protein that is not a minimum in the absence of that molecule)8. Here we show that rotation about covalent bonds in a peptide linker can change a flexible MOF to afford nine distinct crystal structures, revealing a conformational energy landscape that is characterized by multiple structural minima. The uptake of small-molecule guests by the MOF can be chemically triggered by inducing peptide conformational change. This change transforms the material from a minimum on the landscape that is inactive for guest sorption to an active one. Chemical control of the conformation of a flexible organic linker offers a route to modifying the pore geometry and internal surface chemistry and thus the function of open-framework materials.

Reversibly Tuning the Viscosity of Peptide-Based Solutions Using Visible Light.

Light can be used to design stimuli-responsive systems. We induce transient changes in the assembly of a low molecular weight gelator solution using a merocyanine photoacid. Through our approach, reversible viscosity changes can be achieved via irradiation, delivering systems where flow can be controlled non-invasively on demand.

Metal Cross-Linked Supramolecular Gel Noodles: Structural Insights and Antibacterial Assessment.

Achieving precise control over gelator alignment and morphology is crucial for crafting tailored materials and supramolecular structures with distinct properties. We successfully aligned the self-assembled micelles formed by a functionalized dipeptide 2NapFF into long 1-D "gel noodles" by cross-linking with divalent metal chlorides. We identify the most effective cross-linker for alignment, enhancing mechanical stability, and imparting functional properties. Our study shows that Group 2 metal ions are particularly suited for creating mechanically robust yet flexible gel noodles because of their ionic and nondirectional bonding with carboxylate groups. In contrast, the covalent nature and high directional bonds of d-block metal ions with carboxylates tend to disrupt the self-assembly of 2NapFF. Furthermore, the 2NapFF-Cu noodles demonstrated selective antibacterial activity, indicating that the potent antibacterial property of the copper(II) ion is preserved within the cross-linked system. By merging insights into molecular alignment, gel extrusion processing, and integrating specific functionalities, we illustrate how the versatility of dipeptide-based gels can be utilized in creating next-generation soft materials.

Correction: Multi-layer 3D printed dipeptide-based low molecular weight gels.

Correction for 'Multi-layer 3D printed dipeptide-based low molecular weight gels' by Max J. S. Hill et al., Soft Matter, 2022, 18, 5960-5965, https://doi.org/10.1039/D2SM00663D.

Photothermal Perylene Bisimide Hydrogels.

Gels formed using a perylene bisimide (PBI) as a low molecular weight gelator can show the photothermal effect. Formation of the PBI radical anion results in new absorption bands forming, meaning that subsequent irradiation with a wavelength of light overlapping with the new absorption band leads to heating of the gel. This approach can be used to heat the gel, as well as the surrounding milieu. We show how we can use electrochemical methods as well as multicomponent systems to form the radical anion without the need for UV light, and how we can use the photothermal effect to induce phase transitions in the solutions above the gels by exploiting photothermal behavior.

Effect of Imposing Spatial Constraints on Low Molecular Weight Gels.

We outline the effect of imposing spatial constraints during gelation on hydrogels formed by dipeptide-based low molecular weight gelators. The gels were formed via either a solvent switch or a change in pH and formed in different sized vessels to produce gels of different thickness while maintaining the same volume. The different methods of gelation led to gels with different underlying microstructure. Confocal microscopy was used to visualize the resulting microstructures, while the corresponding mechanical properties were probed via cavitation rheology. We show that solvent-switch-triggered gels are sensitive to imposed spatial constraints, in both altered microstructure and mechanical properties, while their pH-triggered equivalents are not. These results are significant because it is often necessary to form gels of different thicknesses for different analytical techniques. Also, gels of different thicknesses are utilized between various applications of these materials. Our data show that it is important to consider the spatial constraints imposed in these situations.

Self-Sorting in Diastereomeric Mixtures of Functionalized Dipeptides.

Self-sorting in functionalized dipeptide systems can be driven by the chirality of a single amino acid, both at a high pH in the micellar state and at a low pH in the gel state. The structures formed are affected to some degree by the relative concentrations of each component showing the complexity of such an approach. The structures underpinning the gel network are predefined by the micellar structures at a high pH. Here, we describe the systems prepared from two dipeptide-based gelators that differ only by the chirality of one of the amino acids. We provide firm evidence for self-sorting in the micellar and gel phases using small-angle neutron scattering and cryo-transmission electron microscopy (cryo-TEM), showing that complete self-sorting occurs across a range of relative concentrations.

Investigating multigelator systems across multiple length scales.

Preparation of multicomponent systems provides a method for changing the properties of low molecular weight gelator (LMWG)-based systems. Here we have prepared a variety of multicomponent systems where both components are N-functionalised dipeptide-based LMWGs that may either co-assemble or self-sort when mixed. We exemplify how varying the concentration ratio of the two components can be used to tune the properties of the multicomponent systems pre-gelation, during gelation and in the gel state using viscosity and rheology measurements, circular dichroism, NMR and small angle neutron scattering. We also investigate the effect of changing the chirality of a single component on the properties of these systems. While predicting the outcome of multicomponent assembly is a challenge, the preparation of a variety of systems allows us to probe the factors affecting their design. This work provides insights into how the properties of multicomponent systems composed of two gelators with the same basic structural design can be tuned by varying the chirality and the concentration ratio of the two components and considering the behaviour of the two components when alone.

Controlled Annealing in Adaptive Multicomponent Gels.

We use a pH-driven annealing process to convert between co-assembled and self-sorted networks in multicomponent gels. The initially formed gels at low pH are co-assembled, with the two components coexisting within the same self-assembled structures. We use an enzymatic approach to increase the pH, resulting in a gel-to-sol transition, followed by a hydrolysis to lower the pH once again. As the pH decreases, a self-sorted network is formed by a two-stage gelation process determined by the pKa of each component. This approach can be expanded to layered systems to generate many varied systems by changing composition and rates of pH change, adapting their microstructure and so allowing access to a far greater range of morphologies and complexity than can be achieved in single component systems.

Personal Perspective on Understanding Low Molecular Weight Gels.

Low molecular weight gels are formed by the self-assembly of small molecules into anisotropic structures that form a network capable of immobilizing the solvent. Such gels are common, with a huge number of different examples existing, and they have many applications. However, there are still significant gaps in our understanding of these systems and challenges that need to be addressed if we are to be able to fully design such systems. Here, a number of these challenges are discussed.

A Self-Assembling Flavin for Visible Photooxidation.

A new flavin-based gelator is reported which forms micellar structures at high pH and gels at low pH. This flavin can be used for the photooxidation of thiols under visible light, with the catalytic efficiency being linked to the self-assembled structures present.

Synthesis and characterisation of diketopyrrolopyrrole-based hydrogels.

Diketopyrrolopyrrole (DPP) based materials can be easily tuned by functionalising with groups that extend the conjugation and thus alter the electronic properties. When attaching thiophenes to give dithiophene-diketopyrrolopyrroles (DTDPPs), a donor-acceptor-donor system is created that is suitable for charge-transfer applications. This core also promotes π-stacking and hydrophobic interactions. Here, we describe a number of DTDPPs functionalised with amino acids that undergo pH-trigerred gelation. We show that the optical properties of our DTDPPs are affected by whether the amino acids have aromatic or aliphatic side chains. We also describe the effect of solvent polarity. We have successfully produced hydrogels via a pH trigger with examples containing phenylalanine (F), valine (V), leucine (L) and alanine (A) amino acids. Viscosity and small angle X-ray scattering measurements show the presence of micellar structures in solution in water at pH 10.5, with gelation starting at a pH less than 7 due to the formation of a fibrous network.

Using small angle scattering to understand low molecular weight gels.

The material properties of a gel are determined by the underpinning network that immobilises the solvent. When gels are formed by the self-assembly of small molecules into a so-called low molecular weight gel, the network is the result of the molecules forming one-dimensional objects such as fibres or nanotubes which entangle or otherwise cross-link to form a three-dimensional network. Characterising the one-dimensional objects and the network is difficult. Many conventional techniques rely on drying to probe the network, which often leads to artefacts. An effective tool to probe the gel in the solvated state is small angle scattering. Both small angle X-ray scattering (SAXS) and small angle neutron scattering (SANS) can be used. Here, we discuss these approaches and provide a tutorial review to describe how these approaches work, what opportunities there are and how the data treatment should be approached. We aim to show the power of this approach and provide enabling information to make them accessible to the non-specialist.

Multi-layer 3D printed dipeptide-based low molecular weight gels.

We describe the direct 3D printing of dipeptide hydrogels, forming layers from gels prepared from different dipeptides. The dipeptides self-assemble into fibres that lead to very different microstructures letting us differentiate between the gels. We show how the mechanical properties of the overall 3D printed structures are affected by the composition of each of the layers, allowing us to build up structures with different microstructure and stiffness. We also discuss the interface between layers formed from different gelators, showing that the gels remain independent from neighbouring printed material, even when prepared in very close proximity.

Electrofabrication of large volume di- and tripeptide hydrogels via hydroquinone oxidation.

The fabrication of protected peptide-based hydrogels on electrode surfaces can be achieved by employing the electrochemical oxidation of hydroquinone to benzoquinone, liberating protons at the electrode-solution interface. The localised reduction in pH below the dipeptide gelator molecules pKa initiates the neutralisation, self-assembly and formation of self-supporting hydrogels exclusively at the electrode surface. Previous examples have been on a nanometre to millimetre scale, using deposition times ranging from seconds to minutes. However, the maximum size to which these materials can grow and their subsequent mechanical properties have not yet been investigated. Here, we report the fabrication of the largest reported di- and tri-peptide based hydrogels using this electrochemical method, employing deposition times of two to five hours. To overcome the oxidation of hydroquinone in air, the fabrication process was performed under an inert nitrogen atmosphere. We show that this approach can be used to form multilayer gels, with the mechanical properties of each layer determined by gelator composition. We also describe examples where gel-to-crystal transitions and syneresis occur within the material.

Correction: Fmoc-diphenylalanine hydrogels: understanding the variability in reported mechanical properties.

Correction for 'Fmoc-diphenylalanine hydrogels: understanding the variability in reported mechanical properties' by Jaclyn Raeburn et al., Soft Matter, 2012, 8, 1168-1174, https://doi.org/10.1039/C1SM06929B.

Spatiotemporal Control Over Base-Catalyzed Hydrogelation Using a Bilayer System.

Controlling the formation and directional growth of hydrogels is a challenge. In this paper, a new methodology to program the gel formation both over space and time is proposed, using the diffusion and subsequent hydrolysis of 1,1'-carbonyldiimidazole from an immiscible organic solution to the aqueous gel media.

Stimuli responsive dynamic transformations in supramolecular gels.

Supramolecular gels are formed by the self-assembly of small molecules under the influence of various non-covalent interactions. As the interactions are individually weak and reversible, it is possible to perturb the gels easily, which in turn enables fine tuning of their properties. Synthetic supramolecular gels are kinetically trapped and usually do not show time variable changes in material properties after formation. However, such materials potentially become switchable when exposed to external stimuli like temperature, pH, light, enzyme, redox, and chemical analytes resulting in reconfiguration of gel matrix into a different type of network. Such transformations allow gel-to-gel transitions while the changes in the molecular aggregation result in alteration of physical and chemical properties of the gel with time. Here, we discuss various methods that have been used to achieve gel-to-gel transitions by modifying a pre-formed gel material through external perturbation. We also describe methods that allow time-dependent autonomous switching of gels into different networks enabling synthesis of next generation functional materials. Dynamic modification of gels allows construction of an array of supramolecular gels with various properties from a single material which eventually extend the limit of applications of the gels. In some cases, gel-to-gel transitions lead to materials that cannot be accessed directly. Finally, we point out the necessity and possibility of further exploration of the field.

Controlling Syneresis of Hydrogels Using Organic Salts.

Supramolecular hydrogels can spontaneously undergo syneresis through fibre-fibre interactions and expel significant amounts of water upon aging. In this process, the hydrophobicity of fibres which regulates the 3D-rearrangement of the self-assembled structures during syneresis is important. Here, we show that we can control the hydrophobic microenvironment of gels by incorporating organic salts into the co-assembled gel fibres thereby enabling control of the macroscopic gel volume phase transition.