Mimicking the extracellular matrix to have a similar nanofibrous structure regarding electrical conductivity and mechanical properties would be highly beneficial for cardiac tissue engineering. The molybdenum disulfide, MoS 2 , and reduced graphene oxide, rGO, nanosheets are two-dimensional nanomaterials which can be considered as great candidates for enhancing the electrical and mechanical properties of biological scaffolds for cardiac tissue engineering applications. In this study, MoS 2 and rGO nanosheets were synthesized and incorporated into silk fibroin nanofibers, SF, via electrospinning method. Then, the human iPSCs transfected with TBX-18 gene, TBX18-hiPSCs, were seeded on these scaffolds for in vitro studies. The MoS 2 and rGO nanosheets were studied by Raman spectroscopy. After incorporation of the nanosheets into SF nanofibers, the associated characterizations were carried out including scanning electron microscopy, transmission electron microscopy, water contact angle, and mechanical test. Furthermore, SF, SF/MoS 2 , and SF/rGO scaffolds were used for in vitro studies. Herein, the scaffolds exhibited acceptable biocompatibility and considerable attachment to TBX18-hiPSCs confirmed by 3-(4, 5dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide, MTT, assay, and scanning electron microscopy. Also, the real-time PCR and immunostaining studies confirmed the maturity and upregulation of cardiac functional genes, including GATA-4, c-TnT, and α-MHC in the SF/MoS 2 and SF/rGO scaffolds compared with the bare SF one.Therefore, the reinforcement of these SF-based scaffolds with MoS 2 and rGO endues them as a suitable candidate for cardiac tissue engineering.