Morphological and Molecular Analysis of Osteoblasts Differentiated from Mesenchymal Stem Cells in Polycaprolactone/Magnesium Oxide/Graphene Oxide Scaffold

Background: The loss or dysfunction of bone tissue that observed after bone tumor resections and severe nonunio‎n fractures afflicts 200 million people worldwide. Bone tissue engineering is a promising approach to repair osteoporotic fractures. Objective: In this paper, polycaprolactone (PCL)/magnesium oxide (MgO)/graphene oxide (GO) nanofibrous scaffold was fabricated by electrospining method, and its biocompatibility and osteogenic differentiation of adipose-derived mesenchymal stem cells (MSCs) on this scaffold were evaluated and compared with pure PCL nanofibrous scaffold. Methods: SEM analysis, DAPI staining and MTT assay were used to evaluation biocompatibility of PCL/ MgO/GO composite scaffold. In addition by ALP assay and proteomic approach, osteostimulatory effect of electrospun composite scaffold was investigated and the expression level of osteogenic markers including Runt-related transcription factor cbfa1/runx2 (runx2), collagen type I (Col1a1) and osteopontin (OPN) in MSCs seeded on PCL/MgO/GO composite scaffold was determined and compared with pure PCL scaffold. Then, RT-PCR technique was used to validate the level expression of these genes. Results: The obtained results showed that adhesion, viability and ALP activity of MSCs on PCL/MgO/GO scaffold considerably enhanced compared with pure PCL. As well as proteomic and real-time analysis illustrated the expression of osteogenic markers including runx2, Col1a1 and OPN increased (>2-fold) in cells seeded on PCL/MgO/GO composite scaffold. Conclusion: It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.