Tissue inhibitors of metalloproteinases level and collagenase activity in gingival crevicular fluid: the relevance to periodontal diseases.
OBJECTIVE
To provide an overall assessment of levels of tissue inhibitors of metalloproteinases (TIMPs), collagenase activities, and of immuno-reactivities for matrix metalloproteinases (MMP)-1 and -8 in gingival crevicular fluid (GCF) obtained from healthy subjects, and gingivitis and periodontitis patients, and to analyse the relationships between periodontal tissue destruction and the GCF components in periodontal diseases by principal component analysis.


METHODS
GCF was sampled with sterile paper strips from 10 gingivitis and 11 periodontitis patients. Ten volunteers served as clinically healthy controls. TIMP-1 and -2 protein amounts in GCF were measured by ELISA, and active and APMA-activatable collagenase activities were determined by functional assays using image-analysis after SDS-PAGE.


RESULTS
GCF TIMP-1 level and both active and latent collagenase activities were significantly higher in the diseased groups than in the healthy group. TIMP-2 was detectable in only 29% of all subjects (mean: 2.06 ng). Western blot analysis showed that MMP-8 was the major interstitial collagenase in the GCF of the diseased groups. Principal component analysis using clinical parameters and the GCF components has indicated components one to three account for 87% of total variation when evaluating the relevance of their measurements to periodontal diseases.


CONCLUSIONS
We conducted the functional and immunological characterization of MMPs and TIMPs in the GCF of periodontally diseased patients. Principal component analysis indicated components one to three explaining 87% of total variation, and further suggested that higher collagenase activity (especially in active collagenase) would be an important marker in evaluating the pathogenesis of periodontitis. Consequently, these observations may have significant therapeutic and diagnostic implications.
Nomura
T
T
Department of Periodontology, Niigata University School of Dentistry, Japan.
Ishii
A
A
Oishi
Y
Y
Kohma
H
H
Hara
K
K
D000328:Adult; D015415:Biomarkers; D015153:Blotting, Western; D016022:Case-Control Studies; D017364:Collagenases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005883:Gingival Crevicular Fluid / Q000201:enzymology*; D005891:Gingivitis / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D020781:Matrix Metalloproteinase 1; D020784:Matrix Metalloproteinase 8; D008875:Middle Aged; D015999:Multivariate Analysis; D010510:Periodontal Diseases / Q000201:enzymology*; D010518:Periodontitis / Q000201:enzymology; D011480:Protease Inhibitors / Q000032:analysis / Q000378:metabolism*; D018709:Statistics, Nonparametric; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis / Q000378:metabolism; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis / Q000378:metabolism*
D015415:Biomarkers; D011480:Protease Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D017364:Collagenases; D020784:Matrix Metalloproteinase 8; D020781:Matrix Metalloproteinase 1