Evaluating the Osteogenic Potential of Reduced Graphene Oxide-Reinforced Poly(3-Hydroxybutyrate)-Chitosan Composite Scaffold for Bone Tissue Regeneration Applications

Abstract

This study aimed to apply electrospinning techniques to design and engineer an optimal scaffold, which is one of the main challenges in bone tissue engineering (BTE). The effects of L-ascorbic acid-reduced graphene oxide on the physicochemical, mechanical, and biological properties of poly(3-hydroxybutyrate)-chitosan (PHB-Cs) scaffolds were investigated. Our findings demonstrate a decrease in fiber diameter, an increased degree of crystallinity, improved hydrophilicity and mechanical strength, and a suitable degradation rate in the presence of rGO reinforcement. XRD and EDS analyses confirmed the formation of lamellar apatite crystals in the bioactivity test. In vitro experiments were performed to assess the biochemical properties of the MG-63 cells. The results from the MTT assay revealed that the mitochondrial activity of cells cultured in both experimental groups exceeded a threshold of 80. Furthermore, the PHB-Cs/rGO composite scaffolds exhibited the highest ALP activity and cellular proliferation rates. The rGO-reinforced composite scaffold promoted the differentiation of MG-63 cells into the osteoblast lineage by upregulating mRNA expression of bone-related genes (Runx2, COL-I, and OCN). In summary, rGO-reinforced composite scaffolds can modulate cellular interactions to restore and enhance bone tissue function, thereby exhibiting potential applications in BTE.