title: MMP-1 (collagenase-1) and MMP-13 (collagenase-3) differentially regulate markers of osteoblastic differentiation in osteogenic cells.
abstract: Previous studies have demonstrated an inverse relationship between constitutive or stimulated collagenase expression and osteoblastic phenotype of osteogenic cells. However, the direct effects of cell-secreted collagenases on osteoblastic differentiation, and the precise contributions of the key collagenolytic MMPs, MMP-1 and -13 to the modulation of specific osteoblastic markers have not been elucidated. Early passage osteogenic human periodontal ligament (PDL) cells were exposed to exogenous collagenase-1 in the presence and absence of dexamethasone. Alternatively, endogenous collagenases were modulated by transfecting the cells with cDNA or siRNA to MMP-1 and/or -13. Specific osteoblastic markers and collagenase expression and activity were then assayed. Increasing concentrations of exogenous collagenase or endogenous MMP-1 and -13 produced a dose-dependent decrease in AP activity. Conversely, a dose-dependent increase in AP activity was observed with increasing concentrations of MMP-1 or MMP-13 siRNA. Overexpression of MMP-1 resulted in a significant decrease in Runx2, osteonectin (ON), osteopontin (OP), bone sialoprotein (BSP) and osteocalcin (OC), but an increase in osterix (Osx) mRNA levels. In contrast, knockdown of MMP-1 caused a significant increase in Runx2, ON, OP, BSP and OC levels and a decrease in Osx levels. MMP-13 overexpression resulted in diminished levels of Osx, OP and BSP, while its knockdown caused a significant increase in Osx and OP levels and a significant decrease in ON levels. The accretion of matrix molecules including collagen I(alpha1) in cell-matrix extracts paralleled the changes in their respective mRNAs. Simultaneous suppression of both MMP-1 and -13 resulted in significant increases in all osteoblastic markers assayed. MMP-1 and -13 differentially regulate osteoblastic markers and their combined suppression is important for the elaboration of an osteoblastic phenotype in PDL cells.
authors.affiliation: The University of Michigan, 1011 North University Avenue, Ann Arbor, Michigan 48109, United States. thayami@umich.edu
authors.forename: Takayuki
authors.initials: T
authors.lastname: Hayami
authors.forename: Yvonne L
authors.initials: YL
authors.lastname: Kapila
authors.forename: Sunil
authors.initials: S
authors.lastname: Kapila