Integrative network pharmacology and experimental validation of rutin-infused collagen-hydroxyapatite scaffold for promoting osteochondral regeneration.

Osteochondral defects (OCD), characterized by bone fractures and cartilage degradation, remain a significant health concern worldwide. This study reports on the development and evaluation of a bioactive scaffold composed of rutin, collagen, and hydroxyapatite (RUT-COL-HAP) for osteochondral tissue regeneration. Network pharmacology identified 51 common target genes of rutin related to chondrocyte and macrophage functions, with ten hub genes playing key roles in inflammation, ECM remodeling, and cell differentiation. Molecular docking revealed strong binding affinities between rutin and its targets. The structural and chemical properties of the scaffolds were assessed using SEM (scanning electron microscopy), FTIR (Fourier transform infrared spectroscopy), TGA (thermogravimetric analysis), and XRD (X-ray diffraction). The results confirmed the successful integration of all the components and demonstrated enhanced crystallinity, thermal stability, and a highly porous, interconnected architecture, particularly in the RUT5-COL-HAP variant. Porosity analysis showed an increase of 50% with higher rutin incorporation, while swelling and biodegradation assessments revealed that RUT5-COL-HAP maintained structural integrity and exhibited controlled degradation over 14 days. In vitro biocompatibility using MG-63 cells demonstrated over 120% cell viability at 72 h, and zebrafish embryo toxicity assays confirmed rutin's safety across all tested concentrations. Overall, the RUT5-COL-HAP scaffold demonstrated high porosity, favorable swelling behavior, and appropriate biodegradation rates. Structural analyses confirmed the ability of rutin to mimic the native extracellular matrix, and toxicity studies verified the safety of rutin for biomedical applications. Thus, it offers a promising biomaterial for osteochondral tissue engineering, providing an effective strategy for treating OCD and improving the clinical outcomes.