Enamel matrix derivative induces the expression of tissue inhibitor of matrix metalloproteinase-3 in human gingival fibroblasts via extracellular signal-regulated kinase.
OBJECTIVE
Periodontal disease is characterized by increased expression and activity of matrix metalloproteinases (MMPs) and insufficient expression/activity of their inhibitors, tissue inhibitors of matrix metalloproteinases (TIMPs). This altered MMP-TIMP balance results in progressive destruction of gingival and periodontal extracellular matrix. Enamel matrix derivative (EMD), clinically used for periodontal regeneration in a device called Emdogain, has been suggested to enhance gingival healing following periodontal procedures in humans. We previously showed that EMD increases the proliferation of human and rat gingival fibroblasts and protects them from tumor necrosis factor-induced apoptosis. In the present study, the modulation of MMP and TIMP expression by EMD was investigated.


METHODS
Primary human gingival fibroblasts were treated in vitro with tumor necrosis factor, EMD or both in serum-free conditions, and RNA was analyzed with an extracellular matrix-focused microarray and quantitative real-time polymerase chain reaction.


RESULTS
Microarray analysis showed detectable expression of MMP-1, MMP-2, MMP-3, MMP-7 and MMP-13, as well as TIMP-1 and TIMP-3 in untreated cells. There was no apparent regulation of the expression of MMP-2, MMP-7, MMP-13 and TIMP-1 by either tumor necrosis factor or EMD. In contrast, tumor necrosis factor significantly increased MMP-1 expression, and EMD reduced it when both agents were present. Also, EMD significantly induced TIMP-3 expression, an effect which was dependent on activation of extracellular signal-regulated kinase 1/2, since it was totally abolished by a selective extracellular signal-regulated kinase pathway inhibitor.


CONCLUSIONS
These data suggest that EMD may affect gingival health by ways other than cell proliferation/survival, i.e. by stimulation of TIMP-3 production, which could improve the MMP-TIMP balance in gingival tissue and curb extracellular matrix destruction.
Zeldich
E
E
Department of Oral Biology, Goldschleger School of Dental Medicine, Tel-Aviv University, Tel-Aviv, Israel.
Koren
R
R
Dard
M
M
Weinberg
E
E
Weinreb
M
M
Nemcovsky
C E
CE
D002070:Butadienes / Q000494:pharmacology; D002478:Cells, Cultured; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology*; D005881:Gingiva / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000187:drug effects; D053509:Matrix Metalloproteinase 13 / Q000187:drug effects; D020778:Matrix Metalloproteinase 2 / Q000187:drug effects; D019278:Matrix Metalloproteinase 3 / Q000187:drug effects; D020783:Matrix Metalloproteinase 7 / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000187:drug effects; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000187:drug effects; D009570:Nitriles / Q000494:pharmacology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000187:drug effects; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D002070:Butadienes; D003746:Dental Enamel Proteins; D004791:Enzyme Inhibitors; D018836:Inflammation Mediators; D009570:Nitriles; C088390:TIMP3 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019717:Tissue Inhibitor of Metalloproteinase-3; D014409:Tumor Necrosis Factor-alpha; C113580:U 0126; C109069:enamel matrix proteins; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1