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Effects of Interleukin‐1β on Matrix Metalloproteinase‐3 Levels in Human Periodontal Ligament Cells
Author(s) -
Nakaya H.,
Oates T.W.,
Hoang A.M.,
Kamoi K.,
Cochran D.L.
Publication year - 1997
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.1997.68.6.517
Subject(s) - periodontal fiber , matrix metalloproteinase , microbiology and biotechnology , messenger rna , polyclonal antibodies , extracellular matrix , matrix metalloproteinase 3 , metalloproteinase , interleukin , reverse transcription polymerase chain reaction , in vitro , chemistry , tissue inhibitor of metalloproteinase , cell culture , andrology , biology , antibody , immunology , cytokine , medicine , biochemistry , gene , dentistry , genetics
M atrix metalloproteinase ‐3 (MMP‐3), or stromelysin‐1, is an enzyme responsible for the degradation of a wide range of extracellular matrix proteins. Increases in MMP‐3 activity have been found in several chronic inflammatory diseases, and this increased activity is thought to be mediated by interleukin‐1β (IL‐1β). Because IL‐1β has been strongly associated with inflammatory periodontal disease, the purpose of this in vitro study was to investigate the role of IL‐1β on the regulation of MMP‐3 levels in cells derived from the human periodontal ligament (PDL). Human PDL cell cultures were treated with IL‐1β at varying concentrations (0.01‐1.0 ng/ml) for 24 hours prior to analysis at either transcript or protein levels. Following the isolation of total RNA, the relative levels of MMP‐3 mRNA were determined using reverse transcription‐polymerase chain reaction (RT‐PCR) with 32P‐end‐labeled primers. Immunocytochemical detection of MMP‐3 protein was performed using polyclonal antibodies to human MMP‐3. The results of RT‐PCR analysis demonstrated a concentrationdependent increase in MMP‐3 mRNA expression, with IL‐1β treatments of 0.1 and 1.0 ng/ml significantly ( P < 0.01) increased over those cells not treated with IL‐1β. This increase in mRNA expression was paralleled by significant ( P < 0.001) changes at the protein level, with an average of 27.6% of the cells stained positive for MMP‐3 following IL‐1β treatment (1.0 ng/ml), compared with control cells showing no positive staining for MMP‐3. In conclusion, the results of this study demonstrate that IL‐1β upregulates MMP‐3 in human PDL cells on both an mRNA and a protein level. These findings suggest possibly important roles for IL‐1β and MMP‐3 in both normal turnover and maintenance of the PDL and in the connective tissue degradation associated with periodontal disease. J Periodontol 1997;68:517–523 .