Objective To investigate the changes in serum tenascin-C (TN-C), soluble CD40 ligand (sCD40L), and secreted phosphoprotein 24 (SPP24) in patients undergoing maintenance hemodialysis (MHD) and their relationship with cardiac valve calcification (CVC).  Methods  A total of 160 MHD patients treated at Hebei PetroChina Central Hospital from January 2022 to May 2025 were selected as the study subjects. General data were collected, and cardiac ultrasound examinations were performed. According to the occurrence of CVC, the patients were divided into a CVC group and a non-CVC group. The general data and serum levels of TN-C, sCD40L, and SPP24 were compared between the two groups. The influencing factors for CVC in MHD patients were analyzed, and receiver operating characteristic (ROC) curves were constructed to evaluate the efficacy of serum TN-C, sCD40L, and SPP24 levels in predicting CVC in MHD patients.  Results  Among the MHD patients included in this study, 83 (51.88%) developed CVC. Compared with the non-CVC group, patients in the CVC group had higher age, dialysis vintage, serum phosphorus, C-reactive protein, TN-C, and sCD40L levels (t=3.188, 2.290, 2.064, 2.049, 5.298, 6.309; P=0.002, 0.023, 0.041, 0.042, ＜0.001, ＜0.001), and lower serum albumin and SPP24 levels (t=2.234, 5.271; P=0.027, ＜0.001). Multivariate logistic regression analysis showed that increased age and elevated serum TN-C and sCD40L levels were risk factors for CVC in MHD patients (OR=1.054, 4.455, 5.512; 95% CI: 1.010～1.101, 1.484～13.377, 1.997～15.215; P=0.016, 0.008, 0.001), while elevated SPP24 levels were a protective factor for CVC in MHD patients (OR=0.322; 95% CI: 0.145～0.713; P=0.005). The AUCs of serum TN-C, sCD40L, and SPP24 alone and in combination for predicting CVC in MHD patients were 0.746, 0.680, 0.752, and 0.858, respectively. The AUC of the combined prediction was higher than that of TN-C, sCD40L, and SPP24 alone (Z=3.333, 4.277, 3.347; all P＜0.001).  Conclusion  In MHD patients with CVC, serum TN-C and sCD40L levels are elevated, while SPP24 levels are decreased. The combined detection of these indicators has high clinical efficacy in predicting CVC in MHD patients.