Photoresponsive Chemistries for User-Directed Hydrogel Network Modulation to Investigate Cell–Matrix Interactions

Synthetic extracellular matrix (ECM) engineering is a highly interdisciplinary field integrating materials and polymer science and engineering, chemistry, cell biology, and medicine to develop innovative strategies to investigate and control cell-matrix interactions. Cellular microenvironments are complex and highly dynamic, changing in response to injury and disease. To capture some of these critical dynamics in vitro, biomaterial matrices have been developed with tailorable properties that can be modulated in situ in the presence of cells. While numerous macromolecules can serve as a basis in the design of a synthetic ECM, our group has exploited multi-arm poly(ethylene glycol) (PEG) macromolecules because of the ease of functionalization, many complementary bio-click reactions to conjugate biological signals, and ultimately, the ability to create well-defined systems to investigate cell-matrix interactions. To date, significant strides have been made in developing bio-responsive and transient synthetic ECM materials that degrade, relax stress, or strain-stiffen in response to cell-mediated stimuli through ECM-cleaving enzymes or integrin-mediated ECM adhesions. However, our group has also designed hydrogels incorporating different photoresponsive moieties, and these moieties facilitate user-defined spatiotemporal modulation of the extracellular microenvironment in vitro. The application of light allows one to break, form, and rearrange network bonds in the presence of cells to alter the biomechanical and biochemical microenvironment to investigate cell-matrix interactions in real-time. Such photoresponsive materials have facilitated fundamental discoveries in the biological pathways related to outside-in signaling, which guide important processes related to tissue development, homeostasis, disease progression, and regeneration. This review focuses on the phototunable chemical toolbox that has been used by Anseth and co-workers to modulate hydrogel properties post-network formation through: bond-breaking chemistries, such as o-nitrobenzyl and coumarin methyl ester photolysis; bond-forming chemistries, such as azadibenzocyclooctyne photo-oligomerization and anthracene dimerization; and bond-rearranging chemistries, such as allyl sulfide addition-fragmentation chain transfer and reversible ring opening polymerization of 1,2-dithiolanes. By using light to modulate the cellular microenvironment (in 2D, 3D, and even 4D), innovative experiments can be designed to study mechanosensing of single cells or multicellular constructs, pattern adhesive ligands to spatially control cell-integrin binding or modulate on-demand the surrounding cell niche to alter outside-in signaling in a temporally controlled manner. To date, these photochemically defined materials have been used for the culture, differentiation, and directed morphogenesis of primary cells and stem cells, co-cultured cells, and even multicellular constructs (e.g., organoids).Herein, we present examples of how this photochemical toolbox has been used under physiological reaction conditions with spatiotemporal control to answer important biological questions and address medical needs. Specifically, our group has exploited these materials to study mesenchymal stem cell mechanosensing and differentiation, the activation of fibroblasts in the context of valve and cardiac fibrosis, muscle stem cell response to matrix changes during injury and aging, and predictable symmetry breaking during intestinal organoid development. The materials and reactions described herein are diverse and enable the design and implementation of an array of hydrogels that can serve as cell delivery systems, tissue engineering scaffolds, or even in vitro models for studying disease or screening for new drug treatments.