The relapse of teeth that have moved during orthodontic treatment is a major clinical issue with respect to the goals of successful treatment. Relaxin has an influence on many physiologic processes, such as collagen turnover. In this study, we determined the effects of relaxin on the relapse and remodeling of periodontal tissue after experimental tooth movement in rats, and we explored the molecular mechanism underlying these processes. To induce experimental tooth movement in rats, 10 g of orthodontic force was applied to the molars. After 14 days, the spring was removed, and then animals began receiving relaxin at a dose of 500 ng/ml for 1 week. The results were evaluated by micro-computed tomography and immunofluorescence staining. In addition, the effects of matrix metalloproteinase (MMP)-1 and MMP-8 production were investigated in human periodontal ligament (hPDL) cells in vitro. The expression of MMP-1 and MMP-8 was analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Furthermore, we demonstrated the signaling pathways involved in relaxin-regulated MMPs expression. The relapse distances and percentages were significantly decreased in the experimental group compared with the controls in vivo. A double-immunofluorescence analysis for Col-I/MMP-1 and Col-I/MMP-8 detected the expression of relaxin in the PDL. Relaxin significantly increased the MMP-1 and MMP-8 expression in a time-dependent manner in hPDL cells in vitro. Furthermore, a p38 inhibitor (SB203580) significantly inhibited the MMP-1 and MMP-8 expression. Our results indicated that relaxin modulates the collagen metabolism, and this hormone may therefore be useful to prevent orthodontic relapse following orthodontic treatment.