Biofabricated genistein incorporated collagen scaffold for enhanced mineralization with refined osteogenic efficacy

Abstract

Bone remodeling is a natural process that involves osteoblasts and osteoclasts. However, this balance can be disrupted due to aging, accidents, and bone defects from conditions like arthritis. Traditionally, these issues have been addressed through metal implantation, tissue grafting, and surgical interventions. However, these methods have limitations, including the risks of inflammation, infection, and delays in donor availability. Bone tissue engineering has become a modern approach to overcoming these challenges. This field utilizes biocompatible and biodegradable biomaterials with osteogenic properties, which reduce the risk of post-surgery infections and inflammation. The development of natural composite scaffolds provides a synergistic effect for osteogenic regeneration. In this study, a collagen-fibrin scaffold incorporated with genistein (Gn) and crosslinked with genipin (Gp), (C-F-Gp-Gn) has been developed to repair osteogenic defects. The binding of genistein to collagen and fibrin was evaluated and verified using Fourier Transform Infrared Spectroscopy. Structural analysis demonstrated that the scaffold’s interconnected pore structure, with a porosity of 60%, is ideal for bone regeneration. Biochemical assays indicated a good swelling potential and an optimal degradation rate of 82% in 14 days. In vitro assays were carried out in MG-63 cells, exhibiting the scaffold’s biocompatibility, while Live/Dead cell staining displayed enhanced cell proliferation and adherence properties. Alizarin Red staining and alkaline phosphatase assays indicated an increased mineralization capacity in the C-F-Gp-Gn scaffold compared to the scaffold without genistein. Gene expression studies showed that the C-F-Gp-Gn complex enhances osteogenic gene expression and improves bone matrix formation. The results suggest that the genistein-incorporated collagen-fibrin scaffold possesses improved osteogenic properties and superior mineralization, making it a promising biomaterial for bone tissue engineering applications.