Nowadays, bone repair by means of stem cells potential is considered as a new approach in regenerative medicine. Adipose-derived mesenchymal stem cells (AD-MSCs) have been investigated as a plentiful cell source with the ability of osteogenic differentiation which can play an important role in bone tissue engineering applications. Discovering proper elements in combination of scaffolds structure to stimulate osteogenesis in adipose-derived stem cells is one of the major concerns in this issue. Porous polymeric scaffolds such as polyacrylonitrile (PAN) and susceptible nanoparticles have attracted a lot of attention recently due to biodegradability and differentiation potential respectively. In the present study, clay-PAN nanocomposite (CPN) and graphene-PAN scaffold have been electrospuned separately and evaluated from the point of the osteogenic potential in AD-MSCs. The objective of this study was to determine the effect of clay and graphene nanoparticles with PAN nanofibers on the fate of viability and osteogenesis of AD-MSCs. First, isolated mesenchymal cells were characterized by flow cytometry. After cell culture on the surface of scaffolds MTT assay, scanning electron microscope (SEM) and DAPI staining were done. The scaffolds were characterized and osteogenic differentiation potential of AD-MSCs has been investigated. The results have indicated that alkaline phosphatase (ALP) activity, calcium content and collagen expression of cells which cultured on clay-PAN nanofibers were higher than cells which cultured on graphene-PAN scaffold. Taken together, these results suggest that porous nanofiber clay-PAN scaffold can enhance the osteogenic differentiation of AD-MSCs, and can be used as a new biodegradable scaffold for bone tissue engineering applications.