Tissue engineering using new strategies has become a growing and promising method for treating large tissue lesions in the body. On the other hand, microRNAs (miRNAs), which are small non-coding regulatory RNAs, are a new class of genetic materials that can have effective pharmacological roles. The combination of these two themes has created promising prospects for the treatment of diseases. Herein, human induced pluripotent stem cells (iPSCs) were transduced with miRNA-2861 and then the osteogenic differentiation potential of transduced iPSCs and non-transduced iPSCs was investigated while cultured on the electrospun poly lactic-co-glycolic acid (PLGA) nanofibrous scaffold and culture plate. MiR-2861-transduced iPSCs showed a significantly higher viability, mineralization, alkaline phosphatase (ALP) activity, calcium content, and bone-related gene expression in comparison with those iPSCs that non-transduced. The results also indicated that this increase is improved when miR-2861 transduced iPSCs are cultured on the PLGA nanofibrous scaffold synergistically. This synergy was also confirmed by the results obtained from of Western blot analysis. It can be concluded that, miR-2861, by negative regulation of those proteins that decrease/inhibit osteogenic differentiation and PLGA nanofibrous scaffold by preparation of a suitable artificial extracellular matrix, have a great positive impact in improving iPSCs osteogenic differentiation potential and this blend can be proposed to use in bone tissue engineering application.