A New Cardiac Decellularized Extracellular Matrix (dECM)-Based Hydrogel: From Its Development with a Standardized Myocardial Decellularization Procedure to In Vitro Model Applications

Cardiovascular diseases remain the leading cause of mortality worldwide, underscoring the urgent need for reliable in vitro models that recapitulate the complexity of the native myocardium. Conventional two-dimensional (2D) cultures lack structural and biochemical complexity, whereas in vivo models are costly, raise ethical concerns, and have poor translational potential. In this study, we developed a novel hydrogel scaffold derived from decellularized porcine ventricular myocardium (dECM). A newly optimized decellularization strategy effectively removed cellular and nuclear components while preserving essential extracellular matrix proteins. The dECM-based hydrogel exhibited reproducible self-crosslinking, gelation kinetics, and stability. Cytocompatibility assays using human bone marrow-derived mesenchymal stem cells demonstrated excellent viability and proliferation upon contact with the biomaterial. Multidimensional hydrogel applications (2.5D and 3D) in vitro revealed higher cell densities than those observed under 2D conditions. Moreover, using human umbilical vein endothelial cells, the dECM-based hydrogel proved to be a valid tool for fabricating cardiovascular in vitro models. As such, this cardiac dECM-based hydrogel is a structurally preserved, biocompatible platform that supports both short- and long-term cell culture. The scaffold has the potential to serve promising applications in cardiac tissue engineering, disease modeling, and cardiotoxicity screening by offering a closer mimicry of the native myocardial environment.