Assessment of a Novel Screening Test for Neutrophil Collagenase Activity in the Diagnosis of Periodontal Diseases

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Abstract
Background: Increased levels of active neutrophil collagenase (MMP‐8) in the gingival crevicular fluid (GCF) are associated with progressive periodontitis. The measurement of this enzyme in GCF could facilitate diagnosis. However, assays with sufficient sensitivity to detect collagenase in whole‐mouth GCF currently use radiolabeled substrates and require several days to complete. To provide more rapid analyses of collagenase activity that are better adapted to clinical studies, we developed and validated a novel assay (soluble biotinylated‐collagen assay: SBA) based on chemiluminescent detection of biotinylated collagen digestion products.Methods: The concordance of the novel SBA assay with a radioactive collagen substrate assay was assessed by parallel analyses of enzyme from 35 neutrophil preparations and from 41 samples of GCF from periodontitis patients, followed by Pearson correlation analysis. To test whether the assay appropriately measured MMP‐8 activity, enzyme activity was assessed after incubation with specific collagenase blockers. We examined the diagnostic utility of the SBA in cross‐sectional and longitudinal analyses of 125 patients with adult periodontitis, 5 patients with early‐onset periodontitis, 1 edentulous patient, and in 32 control patients without periodontitis.Results: The assay detected 18‐fold higher in severe periodontitis than in stable periodontitis and decreased to < 25% of pretreatment levels following therapy. Based on measurements of collagenase activity in different disease groups, we estimated a value of 80 nano units as a threshold for severe periodontitis.Conclusions: These results indicate that active MMP‐8 is detected in GCF by a novel assay that is specific, simple, rapid, and reproducible and which may facilitate diagnostic discrimination between stable and progressive lesions. J Periodontol 1999;70:1292‐1302.

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