Abstract
OBJECTIVE: Delayed fracture healing (DFH) is a major clinical challenge, and sensitive molecular biomarkers remain lacking. This study focused on the regulatory roles of lncRNA DLX6-AS1 and miR-141-3p in DFH. METHODS: A total of 192 patients were enrolled and classified into normal fracture healing (NFH) and DFH groups. Serum samples were analyzed for DLX6-AS1 and miR-141-3p levels via qRT-PCR. Diagnostic value and risk associations were assessed using ROC curves and logistic regression. Human bone marrow mesenchymal stem cells (hBMSCs) were induced to undergo chondrogenic differentiation. Cellular activity including proliferation and apoptosis was measured. Dual-luciferase and rescue assays explored interactions among DLX6-AS1, miR-141-3p, and VEGFA. RESULTS: DLX6-AS1 expression was notably reduced, whereas miR-141-3p was elevated in DFH patients. Both were identified as independent risk factors for DFH. The combined detection of two RNAs demonstrated excellent diagnostic performance. Functional assays revealed that DLX6-AS1 promoted, while miR-141-3p suppressed, hBMSCs proliferation, survival, and chondrogenic differentiation. DLX6-AS1 bound miR-141-3p through a sponging mechanism. miR-141-3p directly targeted VEGFA, which was reduced in DFH and inversely correlated with miR-141-3p abundance. Mechanistic analysis confirmed that the DLX6-AS1/miR-141-3p/VEGFA axis regulates hBMSCs proliferation, apoptosis, and chondrogenic differentiation of fracture healing. CONCLUSION: DLX6-AS1 and miR-141-3p emerge as independent risk indicators and potential diagnostic biomarkers for DFH. By modulating the miR-141-3p/VEGFA axis, DLX6-AS1 influences chondrocyte differentiation and fracture repair, highlighting this pathway as a viable target.