In this study, carboxymethyl chitosan (CMC) was grafted on polycaprolactone (PCL) nanofibers to fabricate scaffolds for bone tissue engineering. The electrospun PCL nanofibers were treated by cold atmospheric plasma (CAP) of helium to generate the reactive functions necessary for CMC grafting. β-carotene (βC) as a biochemical clue and electromagnetic field (EMF, 31.4 μT, 1 h per day) as a biophysical stimulator were used to promote the proliferation and osteodifferentiation of adipose mesenchymal stem cells (ADSCs). Alizarin red staining and calcium content results indicated the generation of nodal calcium on the CMC30%-g-PCL scaffold after 14 days of incubation in the presence or absence of external stimulation factors. Immunocytochemistry (ICC) results confirmed the expression of osteonectin protein for the stem cells seeded on CMC30%-g-PCL with or without using βC or EMF. These results suggest that the fabricated CMC-grafted scaffolds have the ability to self-differentiate stem cells to osteoblasts due to the osteoinductive effects of the grafted CMC. Furthermore, the osteodifferentiation of ADSCs is promoted by using an external stimulation factor such as βC or EMF.
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