Objective To investigate the effect of B-vitamins supplementation on bone turnover biomarkers in hyperhomocysteinemia patients. Methods 96 hyperhomocysteinemia patients were treated with B-vitamins for 4 weeks. The serum levels of total procollagen type 1 amino-terminal propeptide (t-P1NP), β-crosslaps, N-MID osteocalcin, total vitamin D and parathyroid hormone were detected before and after the treatment. Results The t-P1NP level in hyperhomocysteinemia patients was increased significantly (P=0.002) after treatment with B vitamins supplementation. However, there were no significant changes for other four biomarkers before and after treatment. Conclusion After 4 weeks of B-vitamins supplementation, bone formation marker in hyperhomocysteinemia patients is stimulated. Meanwhile, osteocalcin and resorption markers remain the same level as before the treatment.