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In Situ Localization and Characterization of Active Proteases in Chronically Inflamed and Healthy Human Gingival Tissues
Author(s) -
Sarment David P.,
Korostoff Jonathan,
D'Angelo Marina,
Polson Alan M.,
Feldman Roy S.,
Billings Paul C.
Publication year - 1999
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.1999.70.11.1303
Subject(s) - proteases , in situ , gingival and periodontal pocket , medicine , pathology , dentistry , chemistry , periodontal disease , biochemistry , enzyme , organic chemistry
Background: Studies have indicated an important role for host‐derived proteases in the pathogenesis of periodontal disease. The objectives of this study were: 1) to develop an assay measuring protease activity in situ and 2) to localize and characterize the enzymatic activity in intact inflamed and healthy gingiva. Methods: Gingival specimens were prepared and overlaid with a quenched fluorescent substrate. Protease activity was visualized by fluorescence microscopy and correlated with histologic features. Results: In inflamed tissues, enzymatic activity was detected mainly in the connective tissue (predominantly matrix metalloproteases) and, to some extent, in the epithelium (predominantly serine proteases). In contrast, clinically healthy tissues failed to exhibit significant amounts of protease activity. Quantitative and qualitative characteristics of protease activity in intact tissues were found to be pH dependent. Conclusions: The method described here enabled assessment of active proteases in intact tissues where cell‐cell and cell‐matrix interactions had been maintained. Our results indicate that there are substantial differences in the distribution of specific proteases between clinically healthy and inflamed periodontal tissues. J Periodontol 1999;70:1303‐1312.