Elucidating the mechanisms by which acetyl tributyl citrate affects fracture healing: a comprehensive network toxicology study.

BACKGROUND: In this study, we explored the potential risk effects of acetyl tributyl citrate (ATBC) on fracture healing through the method of network toxicology. METHODS: ATBC-related targets were retrieved from the ChEMBL, Swiss Target Prediction, and STITCH databases, whereas fracture healing-related targets were obtained from the GeneCards, OMIM, and GEO databases. Core targets were identified through protein-protein interaction network construction using the STRING database and visualized using the Cytoscape software, followed by GO and KEGG enrichment analyses. Additionally, in-depth analyses, including immune infiltration profiling, gene set enrichment analysis, and ceRNA regulatory network construction, were conducted. Finally, molecular docking and dynamics simulations were performed to determine the binding efficiency and stability for the binding between ATBC and its core targets. RESULTS: A total of 50 overlapping targets were identified, from which six core targets were selected. Using differential expression analysis, five core targets (HDAC2, HDAC3, KAT2B, SMARCA4, and TP53) were further refined. Enrichment analysis suggested potential mechanisms related to the thyroid hormone signaling pathway, the Notch signaling pathway, and the cell cycle. Molecular docking and dynamics simulations confirmed certain binding interactions between ATBC and the core targets. CONCLUSIONS: We provided an integrated network toxicology analysis framework coupled with a molecular dynamics evaluation framework to elucidate the underlying molecular mechanisms through which ATBC interferes with fracture healing, thereby providing novel therapeutic strategies for clinical intervention. However, the findings require further experimental validation and clinical confirmation to establish translational relevance. TRIAL REGISTRATION: Not applicable.