Microassay for the detection of elastase activity in the gingival crevice

J Clin Periodontol. 2001 Jan;28(1):31-7. doi: 10.1034/j.1600-051x.2001.280105.x.

Abstract

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.

Publication types

  • Clinical Trial
  • Randomized Controlled Trial

MeSH terms

  • Adult
  • Clinical Enzyme Tests
  • Coumarins
  • Double-Blind Method
  • Fluorometry / methods
  • Gingival Crevicular Fluid / enzymology*
  • Gingivitis / diagnosis
  • Gingivitis / enzymology*
  • Humans
  • Indicators and Reagents
  • Leukocyte Elastase / analysis*
  • Leukocyte Elastase / metabolism
  • Male
  • Periodontal Index
  • Prospective Studies
  • Regression Analysis
  • Statistics, Nonparametric
  • Substrate Specificity

Substances

  • Coumarins
  • Indicators and Reagents
  • Leukocyte Elastase
  • 7-amino-4-methylcoumarin