Objective  To explore the relationship between serum lipocalin-2 (LCN2), C-C chemokine ligand 3 (CCL3), and C1q tumor necrosis factor-related protein-3 (CTRP3) and vascular calcification (VC) in maintenance hemodialysis (MHD) patients . Methods  Patients who underwent MHD at Hengshui Second People's Hospital from April 2023 to September 2024 were selected as the MHD group. 67 patients who underwent MHD were included as the validation group, and healthy volunteers who underwent physical examinations during the same period were included as the control group. The levels of serum LCN2, CCL3 and CTRP3 were measured by enzyme-linked immunosorbent assay (ELISA). According to the VC status at the last follow-up, patients were divided into the VC group (n=54) and the non-VC group (n=101). Pearson correlation analysis was used to assess correlations; logistic regression was employed to analyze factors influencing the occurrence of VC. The effects of serum phosphorus, calcium, PTH, LCN2, CCL3, and CTRP3 levels on VC were analyzed using relative risk analysis. In addition, the Hayes process analysis program was used to analyze the mediating role of serum indicators in the occurrence of VC. Receiver operating characteristic curve (ROC) curves were used to explore the predictive value of serum LCN2, CCL3 and CTRP3 levels for VC in MHD patients. Results  In this study, 155 patients were included in the MHD group (54 with VC and 101 without VC), and 160 healthy volunteers were included in the control group. Compared with the control group, serum LCN2 (t=26.402) and CCL3 (t=50.873) levels were significantly higher, while CTRP3 level (t=27.085) was lower in the MHD group (all P<0.001). Compared with the non-VC group, serum phosphorus (t=4.132), calcium (t=6.041), parathyroid hormone (PTH, t=20.249) and LCN2 (t=7.172), CCL3 (t=7.553) levels were significantly higher, while CTRP3 level (t=6.533) was significantly lower in the VC group (all P<0.001). Serum phosphorus, calcium, PTH levels were positively correlated with serum LCN2 and CCL3 levels, and negatively correlated with CTRP3 levels (all P<0.001). Multivariate COX regression analysis showed that phosphorus (HR=2.110, 95% CI: 1.158～3.844, P=0.015), calcium (HR=1.677, 95% CI: 1.267～2.496, P=0.011), PTH (HR=2.263, 95% CI 1.197～4.279, P=0.012), LCN2(HR=1.785, 95% CI: 1.106～2.880,P=0.018), and CCL3 (HR=1.562, 95% CI: 1.019～2.395, P=0.041) were independent risk factors for VC, while CTRP3 (HR=0.630, 95% CI: 0.459～0.865, P=0.004) were a protective factor for VC. The risks of developing VC in patients with high expression of phosphorus, calcium, PTH, LCN2, CCL3, and CTRP3 were 2.227 times (95% CI: 1.361～3.641, P<0.001), 2.040 times (95% CI: 1.262～3.298, P=0.002), 2.079 times (95% CI: 1.299～3.327, P=0.001), 1.772 times (95% CI: 1.117～2.811, P=0.012), 4.800 times (95% CI: 2.774～8.305, P=0.002), and 0.581 times (95% CI: 0.369～0.914, P=0.016), respectively, compared with patients with low expression of these biomarkers. The area under the curve (AUC) for predicting VC by serum LCN2, CCL3, and CTRP3 levels were 0.794, 0.831, and 0.849, respectively, and the AUC for the combination of the three was 0.942, with better predictive performance than each individual indicator (Z=4.138, 3.856, 3.523, all P<0.001).  Conclusion   Serum LCN2 and CCL3 levels are elevated, while CTRP3 level is decreased in MHD patients. Combined detection of the three biomarkers may effectively predict the occurrence of VC in MHD patients.