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Microassay for the detection of elastase activity in the gingival crevice
J M Herrmann1, J R Gonzáles, R H Boedeker
1Department of Periodontology, Dental School, University of Giessen, Germany. jens.m.herrmann@dentist.med.uni-giessen.de
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 degrees C, -22 degrees C, -88 degrees C. GCF samples retained elastase activity of almost 100% after a storage of 3 days at -22 degrees 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 microU/microl at baseline to 1444 microU/microl 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.