title: Enamel matrix derivative protein enhances production of matrixmetalloproteinase-2 by osteoblasts.
abstract: BACKGROUND
  Matrix metalloproteinases (MMPs) degrade the extracellular matrix (ECM) and regulate remodeling and regeneration of bone. Enamel matrix derivative (EMD) protein has been used clinically for periodontal regeneration, although its molecular mechanisms are not clear. We evaluated the role of matrix metalloproteinases (MMPs) in regulating EMD-dependent degradation of gelatin on oeoblast-like cell line MG63.
  
  
  METHODS
  MG-63 cells (osteoblast cell line) were incubated with 100 μg/ml EMD protein in the presence or absence of MMP-2 tissue inhibitor for 20 h followed by incubation on DQ-gelatin-coated plates for 4 h. MG-63 cells (1 × 10(6)) were preincubated with SB203580 for 30 min at 37°C and were then placed in 100 μg/ml EMD protein for 24 h. Conditioned media were collected and detected by Western blot analysis.
  
  
  RESULTS
  EMD protein enhanced cell-mediated degradation of gelatin, which was inhibited by the MMP inhibitor TIMP-2. Furthermore, MMP-2 was produced by MG63 cells in response to EMD protein in a P38 MAPK-dependent manner. In addition, blocking of p38 MAPK activation by SB203580 significantly inhibited generation of the active form of MMP-2.
  
  
  CONCLUSIONS
  P38 MAPK pathway promotes expression MMP-2 in EMD activated osteoblasts, which in turn stimulates periodontal regeneration by degrading matrix proteins in periodontal connective tissue.
authors.affiliation: Department of Biochemistry, Osaka Dental University, Osaka, Japan. goda@cc.osaka-dent.ac.jp.
authors.forename: Seiji
authors.initials: S
authors.lastname: Goda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Inoue
authors.forename: Osamu
authors.initials: O
authors.lastname: Takeuchi
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Ujii
authors.forename: Eisuke
authors.initials: E
authors.lastname: Domae
authors.forename: Takashi
authors.initials: T
authors.lastname: Ikeo
mesh_terms: D002460:Cell Line; D017077:Culture Media, Conditioned; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020778:Matrix Metalloproteinase 2 / Q000187:drug effects*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D010006:Osteoblasts / Q000187:drug effects / Q000201:enzymology*; D011725:Pyridines / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D017077:Culture Media, Conditioned; D003746:Dental Enamel Proteins; D004791:Enzyme Inhibitors; D007093:Imidazoles; D061965:Matrix Metalloproteinase Inhibitors; D011725:Pyridines; C560823:TIMP2 protein, human; C109069:enamel matrix proteins; D019716:Tissue Inhibitor of Metalloproteinase-2; D005780:Gelatin; D048051:p38 Mitogen-Activated Protein Kinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C093642:SB 203580