Expression of MMP-13 (collagenase-3) in long-term cultures of human dental pulp cells

Abstract

Matrix metalloproteinase-13 (MMP-13 or collagenase-3) is a member of the family of matrix metalloproteinases (MMPs) produced in high amounts by cells with mineralising potential. Human dental pulp has been shown to express high levels of MMP-13 RNA. Objective Since human dental pulp derived cells (HDPC) are known to possess osteoprogenitor properties, we investigated the pattern of expression of MMP-13 in long-term cultures of those cells under conditions that support mineralisation in vitro. Design Impacted teeth or teeth extracted for orthodontic purposes were used to obtain dental pulp explants and HDPC were cultured for approximately 5 weeks. Pro- and active MMP-13 levels were determined in the cell culture supernatants by means of enzyme-linked immunosorbent assay (ELISA). Cell growth was evaluated through DNA content and osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity and Alizarin Red staining. Results Mineralising cultures of HDPC produced significantly higher levels of pro-MMP-13 compared to control cultures. Both pro- and active MMP-13 levels displayed a characteristic peak that was found to coincide with the peak in alkaline phosphatase activity and the onset of mineralisation. Once mineralisation was firmly established, MMP-13 expression was significantly reduced. Conclusions Evidence from this study suggests a role for MMP-13 in the transition of human dental pulp cells to a mature mineralising phenotype and points to MMP-13 as a possible marker in HDPC differentiation.