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Microassay for the detection of elastase activity in the gingival crevice
Author(s) -
Herrmann Jens Martin,
Gonzáles José Roberto,
Boedeker Rolf Hasso,
Vonholdt Julia,
Meyle Joerg
Publication year - 2001
Publication title -
journal of clinical periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.456
H-Index - 151
eISSN - 1600-051X
pISSN - 0303-6979
DOI - 10.1111/j.1600-051x.2001.280105.x
Subject(s) - elastase , gingivitis , chemistry , enzyme assay , enzyme , medicine , dentistry , biochemistry
Background, aims: A new microassay for the detection of elastase activity (EA) in gingival crevicular fluid (GCF) has been established. GCF was collected with Periopaper ® strips and quantified in a Periotron ® . Methods: Enzyme activity was measured in a microtiter plate reader, using a fluorometric assay. To ensure quality and precision of the assay, recovery rates were determined at different activities with a recovery of >90%. In a 2nd step, stability of the enzyme was investigated during storage at room temperature, +4 °C, −22 °C, −88 °C. GCF samples retained elastase activity of almost 100% after a storage of 3 days at −22 °C. In a group of 12 healthy volunteers, elastase activity was assayed throughout an 18 day experimental gingivitis protocol. Results: Median activity increased from 481 μU/μl at baseline to 1444 μU/μl at day 18, which was accompanied by the development of the signs of gingivitis. The increase of EA during the experimental phase of the study was highly significant ( p <0.001) and correlated well with the increasing severity of gingivitis. Conclusion: The data suggest that elastase activity in GCF is an excellent quantitative measure of gingival inflammation.