Microassay for the detection of elastase activity in the gingival crevice.
OBJECTIVE
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.


CONCLUSIONS
The data suggest that elastase activity in GCF is an excellent quantitative measure of gingival inflammation.
Herrmann
J M
JM
Department of Periodontology, Dental School, University of Giessen, Germany. jens.m.herrmann@dentist.med.uni-giessen.de
Gonzáles
J R
JR
Boedeker
R H
RH
Vonholdt
J
J
Meyle
J
J
D000328:Adult; D004796:Clinical Enzyme Tests; D003374:Coumarins; D004311:Double-Blind Method; D005470:Fluorometry / Q000379:methods; D005883:Gingival Crevicular Fluid / Q000201:enzymology*; D005891:Gingivitis / Q000175:diagnosis / Q000201:enzymology*; D006801:Humans; D007202:Indicators and Reagents; D019272:Leukocyte Elastase / Q000032:analysis* / Q000378:metabolism; D008297:Male; D010512:Periodontal Index; D011446:Prospective Studies; D012044:Regression Analysis; D018709:Statistics, Nonparametric; D013379:Substrate Specificity
D003374:Coumarins; D007202:Indicators and Reagents; D019272:Leukocyte Elastase; C028743:7-amino-4-methylcoumarin