Background <p>Long non-coding RNAs (lncRNAs) exert a vital role in the fracture healing process by interacting with microRNAs (miRNAs) and constructing lncRNA-miRNA regulatory networks. This study focuses on clarifying lncRNA LINC01446’s role in delayed fracture healing (DFH) and exploring its potential regulatory mechanism.</p> Methods <p>The expression of LINC01446 and miR-338-3p in serum samples was detected via qRT-PCR, and the predictive ability of LINC01446 for DFH in fracture patients was evaluated by the ROC curve. The targeting interaction between LINC01446 and miR-338-3p was predicted via StarBase and confirmed through the dual-luciferase reporter assay. For the assessment of cell proliferation and apoptosis, the CCK-8 assay and flow cytometry were utilized.</p> Results <p>In DFH, LINC01446 was significantly downregulated, while miR-338-3p was notably increased. The relationship between LINC01446 and miR-338-3p was identified, showing an inverse regulatory pattern. Knockdown of LINC01446 lowered osteogenic marker gene expression, inhibited the proliferation of MC3T3-E1 cells, and induced cell apoptosis. Additionally, the miR-338-3p inhibitor effectively abrogated the suppressive effects of LINC01446 knockdown on osteoblastic proliferation and differentiation, as well as its promotional effect on cell apoptosis (<i>P</i> &lt; 0.05).</p> Conclusions <p>LINC01446 can affect DFH via miR-338-3p and may act as a potential therapeutic target.</p>