Controlled, supramolecular polymer formulation to engineer hydrogels with tunable mechanical and dynamic properties

Nature uses combined covalent (chemical bonds) and non-covalent (physical bonds) synthesis in a highly, controlled multi-step fashion to create functional materials with different mechanical and dynamic properties out of similar building blocks. Surprisingly, this control in fully synthetic systems remains elusive, even though the effects of formulation pathways on the assembly processes have been emphasized—highlighting the importance of and relationship between energy landscapes and function in synthetic systems. Here, we control multiple, coherent supramolecular assembly processes (fiber formation and crosslinking) to formulate hydrogels with tunable mechanical and dynamic properties. Hydrogels are prepared via two different formulation methods using similar building blocks (monofunctional and bifunctional supramolecular monomers), including (1) the mixing of the supramolecular monomers under basic conditions (Fbasic) and (2) the mixing of the supramolecular monomers under neutral conditions (Fneutral). In Fbasic, network formation is induced via simultaneous fiber formation and crosslinking, yielding homogeneously mixed networks which are dense, stiff (~10 kPa) and robust. In Fneutral, network formation is induced through sequential fiber formation and crosslinking, yielding heterogenous, soft (~2 kPa) and dynamic networks. With these results, we advance towards the controlled, multistep, non-covalent synthesis to create larger hierarchical, functional structures, similar to nature.</p

Development of a Fully Synthetic Corneal Stromal Construct via Supramolecular Hydrogel Engineering

Recent advances in the field of ophthalmology show great potential in the design of bioengineered constructs to mimic the corneal stroma. Hydrogels based on synthetic supramolecular polymers, are attractive synthetic mimics of the natural highly hydrated corneal stroma. Here, a fully synthetic corneal stromal construct is developed via engineering of an injectable supramolecular hydrogel based on ureido-pyrimidinone (UPy) moieties. The hydrogel displays a dynamic and tunable behavior, which allows for control of biochemical and mechanical cues. Two hydrogels are developed, a fully synthetic hydrogel functionalized with a bioactive cyclic arginine-glycine-aspartate UPy (UPy-cRGD) additive, and a hybrid hydrogel based on UPy-moieties mixed with collagen type I fibers. Both hydrogels supported cell encapsulation and associated cellular deposition of extracellular matrix (ECM) proteins after 21 days. Excitingly, the hydrogels support the activation of isolated primary keratocytes into stromal fibroblasts as well as the differentiation toward more quiescent corneal stromal keratocytes, demonstrated by their characteristic long dendritic protrusions and a substantially diminished cytokine secretion. Furthermore, cells survive shear stresses during an injectability test. Together, these findings highlight the development of an injectable supramolecular hydrogel as a synthetic corneal stromal microenvironment able to host primary keratocytes.</p

Virus-like particles as crosslinkers in fibrous biomimetic hydrogels: Approaches towards capsid rupture and gel repair

Biological hydrogels can become many times stiffer under deformation. This unique ability has only recently been realised in fully synthetic gels. Typically, these networks are composed of semi-flexible polymers and bundles and show such large mechanical responses at very small strains, which makes them particularly suitable for application as strain-responsive materials. In this work, we introduced strain-responsiveness by crosslinking the architecture with a multi-functional virus-like particle. At high stresses, we find that the virus particles disintegrate, which creates an (irreversible) mechanical energy dissipation pathway, analogous to the high stress response of fibrin networks. A cooling-heating cycle allows for re-crosslinking at the damaged site, which gives rise to much stronger hydrogels. Virus particles and capsids are promising drug delivery vehicles and our approach offers an effective strategy to trigger the release mechanically without compromising the mechanical integrity of the host material

In Vivo Retention Quantification of Supramolecular Hydrogels Engineered for Cardiac Delivery

Recent advances in the field of cardiac regeneration show great potential in the use of injectable hydrogels to reduce immediate flush-out of injected factors, thereby increasing the effectiveness of the encapsulated drugs. To establish a relation between cardiac function and retention of the drug-encapsulating hydrogel, a quantitative in vivo imaging method is required. Here, the supramolecular ureido-pyrimidinone modified poly(ethylene glycol) (UPy-PEG) material is developed into a bioactive hydrogel for radioactive imaging in a large animal model. A radioactive label is synthesized, being a ureido-pyrimidinone moiety functionalized with a chelator (UPy-DOTA) complexed with the radioactive isotope indium-111 (UPy-DOTA-111In) that is mixed with the hydrogel. Additionally, bioactive and adhesive properties of the UPy-PEG hydrogel are increased by supramolecular introduction of a UPy-functionalized recombinant collagen type 1-based material (UPy-PEG-RCPhC1). This method enables in vivo tracking of the nonbioactive and bioactive supramolecular hydrogels and quantification of hydrogel retention in a porcine heart. In a small pilot, cardiac retention values of 8% for UPy-PEG and 16% for UPy-PEG-RCPhC1 hydrogel are observed 4 h postinjection. This work highlights the importance of retention quantification of hydrogels in vivo, where elucidation of hydrogel quantity at the target site is proposed to strongly influence efficacy of the intended therapy