biostudies-literatureJagiello AEdwards LifesciencesNational Institutes of Health Office of the DirectorNational Institutes of HealthNIH HHSNational Science Foundation39-47https://www.ebi.ac.uk/biostudies/studies/S-EPMC9552552biostudies-literatureUnknown141Cells are known to constantly interact with their local extracellular matrix (ECM) and respond to a variety of biochemical and mechanical cues received from the ECM. Nonetheless, comprehensive understanding of cell-ECM interactions has been elusive. Many studies rely on analysis of cell behavior on 2D substrates, which do not reflect a natural cell environment. Further, lack of dynamic control over local stiffness anisotropies and fiber alignment hinders progress in studies in naturally derived fibrous 3D cultures. Here, we present a cell-safe method of patterned photocrosslinking, which can aid in studying biological hypotheses related to mechanotransduction in 3D hydrogels. As previously described by our group, ruthenium-catalyzed photocrosslinking (RCP) of selected ECM regions promotes Acta biomaterialiaPatterned photocrosslinking to establish stiffness anisotropies in fibrous 3D hydrogels.PMC95525521S10OD025064-01A1S10 OD025064DMS-1953410Botvinick ECastillo UJagiello AHu QfalsePatterned photocrosslinking to establish stiffness anisotropies in fibrous 3D hydrogels.Cells are known to constantly interact with their local extracellular matrix (ECM) and respond to a variety of biochemical and mechanical cues received from the ECM. Nonetheless, comprehensive understanding of cell-ECM interactions has been elusive. Many studies rely on analysis of cell behavior on 2D substrates, which do not reflect a natural cell environment. Further, lack of dynamic control over local stiffness anisotropies and fiber alignment hinders progress in studies in naturally derived fibrous 3D cultures. Here, we present a cell-safe method of patterned photocrosslinking, which can aid in studying biological hypotheses related to mechanotransduction in 3D hydrogels. As previously described by our group, ruthenium-catalyzed photocrosslinking (RCP) of selected ECM regions promotes 2022-01-01T00:00:00Z2022 Mar2025-04-29T11:02:35.534Z2025-04-06T19:48:00.382ZS-EPMC95525523497178610.1016/j.actbio.2021.12.028