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

 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.

 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.

 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.

 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.

 It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.