In this study, ultrafine magnetic nanofibers were developed for bone regeneration purposes. Nanofibers were acquired by electrospinning using a two-component nanofiber matrix (CP: chitosan (Cs) and polyvinyl alcohol (PVA)) containing different concentrations of succinate conjugated-magnetic hydroxyapatite nanocomposites (SMHA). Hybrid nanofibers (CP&SMHA) containing 5 mg ml-1 of SMHA nanocomposite showed well-defined properties in terms of physicochemical properties and cell behavior. Then, they were modified with adenosine 5ʹ-triphosphate (ATP) and Mg2+ ions. The initial adhesion of mesenchymal stem cells and their proliferation rate on the surface of modified nanofibers (Mg.ATP.CP&SMHA) were significantly increased as compared to those of bare nanofibers. Analysis of common osteogenic markers such as alkaline phosphatase activity and the expression of Runt-related transcription factor 2 and osteocalcin confirmed the osteogenic efficacy enhancement of CP&SMHA nanofibers when they were functionalized with ATP and Mg2+. The utilization of the antagonist of purine receptor, P2X7, revealed that this receptor has a major role in the osteogenesis process induced by Mg.ATP.CP&SMHA. Moreover, the results showed that cell adhesion, proliferation, and differentiation improved as nanofibers were under the influence of the electromagnetic field (EMF), displaying synergistic effects in the process of bone formation. Mg.ATP.CP&SMHA also showed an antibacterial effect against gram-negative and gram-positive bacteria, Escherichia coli and Staphylococcus aureus, respectively. Considering the high osteogenic potential and antibacterial activity of Mg.ATP.CP&SMHA nanofibers particularly in combination with EMF, it can serve as a great candidate for use in bone tissue engineering applications.

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