The elastic modulus of the extracellular matrix is a dynamic property that changes during various biological processes, such as disease progression or wound healing. Most cell culture platforms, however, have traditionally exhibited static properties, making it necessary to replate cells to study the effects of different elastic moduli on cell phenotype. Recently, much progress has been made in the development of substrates with mechanisms for either increasing or decreasing stiffness in situ, but there are fewer examples of substrates that can both stiffen and soften, which may be important for simulating the effects of repeated ECM injury and resolution. In the work presented here, poly(ethylene glycol)-based hydrogels reversibly stiffen and soften with multiple light stimuli via photoisomerization of an azobenzene-containing crosslinker. Upon irradiation with cytocompatible doses of 365 nm light (10 mW/cm2, 5 min), isomerization to the azobenzene cis configuration leads to a softening of the hydrogel up to 100-200 Pa (shear storage modulus, G’). This change in gel properties is maintained over a timescale of several hours due to the long half-life of the cis isomer. The initial modulus of the gel can be recovered upon irradiation with similar doses of visible light. With applications in mechanobiology in mind, cytocompatibility with a mechanoresponsive primary cell type is demonstrated. Porcine aortic valvular interstitial cells were encapsulated in the developed hydrogels and shown to exhibit high levels of survival, as well as a spread morphology. The developed hydrogels enable a route to the noninvasive control of substrate modulus independent of changes in the chemical composition or network connectivity, allowing for investigations of the effect of dynamic matrix stiffness on adhered cell behavior.

The relationship between the structure of sequence-defined peptoid polymers and their ability to assemble into well-defined nanostructures is important to the creation of new bioinspired platforms with sophisticated functionality. Here, the hydrophobic N-(2-phenylethyl)glycine (Npe) monomers of the standard nanosheet-forming peptoid sequence were modified in an effort to (1) produce nanosheets from relatively short peptoids, (2) inhibit the aggregation of peptoids in bulk solution, (3) increase nanosheet stability by promoting packing interactions within the hydrophobic core, and (4) produce nanosheets with a nonaromatic hydrophobic core. Fluorescence and optical microscopy of individual nanosheets reveal that certain modifications to the hydrophobic core were well tolerated, whereas others resulted in instability or aggregation or prevented assembly. Importantly, we demonstrate that substitution at the meta and para positions of the Npe aromatic ring are well tolerated, enabling significant opportunities to tune the functional properties of peptoid nanosheets. We also found that N-aryl glycine monomers inhibit nanosheet formation, whereas branched aliphatic monomers have the ability to form nanosheets. An analysis of the crystal structures of several N,N'-disubstituted diketopiperazines (DKPs), a simple model system, revealed that the preferred solid-state packing arrangement of the hydrophobic groups can directly inform the assembly of stable peptoid nanosheets.

Peptoid nanosheets are novel protein-mimetic materials that form from the supramolecular assembly of sequence-defined peptoid polymers. The component polymer chains organize themselves via a unique mechanism at the air-water interface, in which the collapse of a compressed peptoid monolayer results in free-floating, bilayer nanosheets. To impart functionality into these bilayer materials, structural engineering of the nanosheet-forming peptoid strand is necessary. We previously synthesized a series of peptoid analogues with modifications to the hydrophobic core in order to probe the nanosheet tolerance to different packing interactions. Although many substitutions were well-tolerated, routine surface pressure measurements and monolayer collapse isotherms were insufficient to explain which molecular processes contributed to the ability or inability of these peptoid analogues to form nanosheets. Here, we show that surface dilational rheology measurements of assembled peptoid monolayers at the air-water interface provide great insight into their nanosheet-forming ability. We find that a key property required for nanosheet formation is the ability to assemble into a solidlike monolayer in which the residence time of the peptoid within the monolayer is very long and does not exchange rapidly with the subphase. These collapse-competent monolayers typically have a characteristic time of diffusion-exchange values, τ, of >5000 s. Thus, rheological measurements provide an efficient method for assessing the nanosheet-forming ability of peptoid analogues. Results from these studies can be used to guide the rational design of peptoids for assembly into functional nanosheets.

There is a clear link between defects in autophagy and the development of autoimmune and chronic inflammatory diseases, raising interest in better understanding the roles of autophagy within the immune system. In addition, autophagy has been implicated in the immune response to infection by pathogenic microbes. As such, there are efforts currently underway to develop modulators of autophagy as a therapeutic strategy for the treatment of the autoimmune, inflammatory, and infectious diseases. In this review, we discuss the numerous roles for autophagy in immunity and how these activities are linked to disease. We highlight how autophagy affects pathogen clearance, phagocytosis, pattern recognition receptor signaling, inflammation, antigen presentation, cell death, and immune cell development and maintenance. With these diverse and extensive immune-related functions for autophagy in mind, we finish by considering the possible implications of targeting autophagy as a therapeutic strategy.