In the present study, we developed and assessed novel injectable hydrogel matrices combined with nanoparticles and secretome biomolecules to reduce Doxorubicin (DOX)- induced cytotoxicity in human stem cell-derived cardiomyocytes. A Fe2O3 nanoparticle-loaded biocompatible silk sericin (MSS) nanocomposite form was fabricated and used as an injectable carrier for secretome for in vivo cardiomyocyte metabolism. Biological analyses revealed that the secretome-encapsulated Fe3O2-Silk sericin (Sec@MSS) hydrogel markedly reduced acridine orange/ethidium bromide (AO-EB) dual staining in cardiomyocytes, revealing significantly induced apoptosis. Furthermore, we evaluated the mitochondrial membrane potential for DOX and Sec@MSS hydrogel, and demonstrated apoptosis of the cardiomyocytes in the DOX-alone and Sec@MSS groups. However, the cardiotoxicity of Sec@MSS sericin was much lower than that in the DOX group. The acoustic behaviors of the functionalized Sec@MSS were examined. The results indicate that Sec@MSS hydrogel might serve as an effective treatment agent in cardiac diseases in the future.