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


METHODS
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.
Suri
Lokesh
L
Department of Orthodontics, DHS-2, Tufts University, School of Dental Medicine, One Kneeland Street, Boston, MA 02111, USA. lokesh.suri@tufts.edu
Damoulis
Petros D
PD
Le
Trang
T
Gagari
Eleni
E
D000469:Alkaline Phosphatase / Q000032:analysis; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism*; D010012:Osteogenesis / Q000502:physiology
D007770:L-Lactate Dehydrogenase; D000469:Alkaline Phosphatase; D053509:Matrix Metalloproteinase 13