The transient self-assembly of molecules under the direction of a consumable fuel source is fundamental to biological processes such as cellular organization and motility. Such biomolecular assemblies exist in an outof-equilibrium state, requiring continuous consumption of high energy molecules. At the same time, the creation of bioinspired supramolecular hydrogels has traditionally focused on associations occurring at the thermodynamic equilibrium state. Here, hydrogels are prepared from cucurbit[7]uril host-guest supramolecular interactions through transient physical crosslinking driven by the consumption of a reactive chemical fuel. Upon action from this fuel, the affinity and dynamics of CB[7]-guest recognition are altered. In this way, the lifetime of transient hydrogel formation and the dynamic modulus obtained are governed by fuel consumption, rather than being directed by equilibrium complex formation.

The fields of precision imaging and drug delivery have revealed a number of tools to improve target specificity and increase efficacy in diagnosing and treating disease. Biological molecules, such as antibodies, continue to be the primary means of assuring active targeting of various payloads. However, molecularscale recognition motifs have emerged in recent decades to achieve specificity through the design of interacting chemical motifs. In this regard, an assortment of bioorthogonal covalent conjugations offer possibilities for in situ complexation under physiological conditions. Herein, a related concept is discussed that leverages interactions from noncovalent or supramolecular motifs to facilitate in situ recognition and complex formation in the body. Classic supramolecular motifs based on host−guest complexation offer one such means of facilitating recognition. In addition, synthetic bioinspired motifs based on oligonucleotide hybridization and coiled-coil peptide bundles afford other routes to form complexes in situ. The architectures to include recognition of these various motifs for targeting enable both monovalent and multivalent presentation, seeking high affinity or engineered avidity to facilitate conjugation even under dilute conditions of the body. Accordingly, supramolecular "click chemistry" offers a complementary tool in the growing arsenal targeting improved healthcare efficacy.

Host−guest physical cross-linking has been used to prepare supramolecular hydrogels for various biomedical applications. More recent efforts to endow these materials with stimuli-responsivity offers an opportunity to precisely tune their function for a target use. In the context of light-responsive materials, azobenzenes are one prevailing motif. Here, an asymmetric azobenzene was explored for its ability to form homoternary complexes with the cucurbit[8]uril macrocycle, exhibiting an affinity (K eq ) of 6.21 × 10 10 M −2 for sequential binding, though having negative cooperativity. Copolymers were first prepared from different and tunable ratios of NIPAM and DMAEA, and DMAEA groups were then postsynthetically modified with this asymmetric azobenzene. Upon macrocycle addition, these polymers formed supramolecular hydrogels; relaxation dynamics increased with temperature due to temperature-dependent affinity reduction for the ternary complex. Application of UV light disrupted the supramolecular motif through azobenzene photoisomerization, prompting a gel-to-sol transition in the hydrogel. Excitingly, within several minutes at room temperature, thermal relaxation of azobenzene to its trans state afforded rapid hydrogel recovery. By revealing this supramolecular motif and employing facile means for its attachment onto pre-synthesized polymers, the approach described here may further enable stimuli-directed control of supramolecular hydrogels for a number of applications.

Supramolecular hydrogels prepared from host−guest physical cross-linking of polymers have versatile utility in a number of applications. Routes to integrate stimuliresponsive features in these materials are intended to add another dimension to enhance their functionality. Herein, a guest which forms a homoternary complex with the cucurbit[8]uril macrocycle was appended to the ends of Pluronic F-127 polymers. This polymer undergoes temperature-responsive micelle formation, upon which CB [8] promotes their physical cross-linking via its host−guest interactions with the appended guests yielding a percolated hydrogel network. The particular guests used to form the homoternary complex can further be photo-dimerized to replace the physical host−guest interaction with a covalently bonded interaction. This change results in a reduction in hydrogel dynamics of roughly 2 orders of magnitude, yet temperature-responsive gelation and overall network architecture remain apparently unchanged. Hydrogels composed of micelles cross-linked by both supramolecular and photo-dimerized interactions support the injection and encapsulation of cells and enable inclusion and release of macromolecular payloads in vitro and in vivo. Thus, this approach points to a strategy to integrate external stimuli into supramolecular hydrogels through a combination of responsive polymers and light-directed supramolecular motifs.

Hydrogels prepared from supramolecular cross-linking motifs are appealing for use as biomaterials and drug delivery technologies. The inclusion of macromolecules (e.g., protein therapeutics) in these materials is relevant to many of their intended uses. However, the impact of dynamic network cross-linking on macromolecule diffusion must be better understood. Here, hydrogel networks with identical topology but disparate cross-link dynamics are explored. These materials are prepared from cross-linking with host–guest complexes of the cucurbit[7]­uril (CB[7]) macrocycle and two guests of different affinity. Rheology confirms differences in bulk material dynamics arising from differences in cross-link thermodynamics. Fluorescence recovery after photobleaching (FRAP) provides insight into macromolecule diffusion as a function of probe molecular weight and hydrogel network dynamics. Together, both rheology and FRAP enable the estimation of the mean network mesh size, which is then related to the solute hydrodynamic diameters to further understand macromolecule diffusion. Interestingly, the thermodynamics of host–guest cross-linking are correlated with a marked deviation from classical diffusion behavior for higher molecular weight probes, yielding solute aggregation in high-affinity networks. These studies offer insights into fundamental macromolecular transport phenomena as they relate to the association dynamics of supramolecular networks. Translation of these materials from in vitro to in vivo is also assessed by bulk release of an encapsulated macromolecule. Contradictory in vitro to in vivo results with inverse relationships in release between the two hydrogels underscores the caution demanded when translating supramolecular biomaterials into application.