<i>Emerita Analoga</i> Shell-Derived CS/GO Composite Incorporated into a Biomimetic PAN Nanofiber Membrane for Enhanced Bone Tissue Regeneration

Bone regeneration is a process that aims to restore the structure and function of damaged bone tissues. Modern approaches for bone regeneration involve a combination of strategies, including tissue engineering and biomaterials, to promote healing. In this study, electrospun nanofibers were developed by using biosynthesized chitosan (CS)- and graphene oxide (GO)-loaded polyacrylonitrile (PAN) nanofibers. These scaffolds demonstrated stable mechanical support and capability to promote rapid bone defect repair. The physicochemical properties of the prepared nanoparticles and nanofibers were characterized using XRD and XPS analysis. The nanofiber morphology and structure of the CS/GO composite were analyzed through SEM and TEM. In vitro studies and ALP activity demonstrated the membranes capability to promote new bone formation and support healing, and Alizarin red staining highlighted the membrane's ability to enhance cell-cell interactions and increase calcium deposition, crucial for tissue regeneration. Cytotoxicity analysis revealed that 97.66 ± 1.5% of MG-63 cells remained viable on the surface of the prepared nanofiber, as assessed by the MTT assay. At the molecular level, real-time RT-PCR was used to examine the mRNA expression of Runx2 and type 1 collagen. Promoting osteogenic gene expression and enhancing mineral deposition on the prepared nanofiber show significant potential in accelerating bone healing and ensuring the successful integration of the scaffold with the surrounding bone tissue. Based on these findings, we conclude that the CS/GO@PAN nanofibrous membrane holds significant promise as a substrate for bone regeneration.