A relationship between proteinase activity and clinical parameters in the treatment of periodontal disease

Abstract

The objective of this research was to determine the effectiveness of a biochemical assay which measures proteolytic enzyme activity in gingival crevicular fluid (GCF) and to relate this enzyme activity to clinical parameters traditionally utilized for periodontitis detection. A clinical trial was conducted on 8 periodontitis subjects with > or =4 sites exhibiting a loss of attachment of > or =5 mm and probing depths of > or =5 mm with bleeding on probing. On each subject, a plaque index was performed, followed by GCF sampling at those sites which exhibited a loss of attachment and probing depths. GCF was analyzed for activity against benzoyl-L-arginine-p-nitroanilide in the presence (BAPNA w/gly-gly) and the absence (BAPNA w/o gly-gly) of glycyl-glycine and against MeOSuc-Ala-Ala-Pro-Val-pNA and Suc-Ala-Ala-Pro-Phe-pNA for neutrophil serine proteinases activity (elastase and cathepsin G, respectively). Subsequently, a gingival index was performed, attachment levels and probing depths were recorded using a constant force probe with bleeding on probing being noted. A split-mouth design was employed and half mouths were randomly assigned to the following treatment groups: group A, half of the mouth received scaling/root planing and polishing: group B, half of the mouth received no treatment (control). Subjects were treated, then instructed on toothbrushing and interdental cleaning. After 4 weeks, subjects returned to receive a plaque index; GCF sampling, gingival index, attachment levels, probing depths and bleeding on probing as described above. Using a paired Student t-test, the findings suggest that BAPNA w/gly-gly was significantly less in treatment sites than in non-treated control sites (p=0.05). No such correlation was found for other activities, including neutrophil serine proteinases which were shown to occur in GCF in free, proteolytically active forms. In addition, significant treatment effects were detected for probing depths (p= 0.03) which reduced by 1.3 mm and attachment levels (p=0.02) which gained 0.7 mm. The reduction of P. gingivalis from treated periodontitis sites as detected by a significant decrease in BAPNA w/ gly-gly may prove to be a valuable marker for periodontal disease activity.

Activation of Human Oral Epithelial Cells by Neutrophil Proteinase 3 Through Protease-Activated Receptor-2

“…Infiltration of neutrophils into gingival tissues is an early event in gingival inflammation, and neutrophils are the predominant leukocytes in the gingival crevicular fluid (GCF) (ϳ90%) (48). It is also evident that GCF has neutrophil serine proteinase activities against MeOSuc-Ala-Ala-Pro-Val-pnitroanilide for HLE and Suc-Ala-Ala-Pro-Phe-p-nitroanilide for Cat G (49). Because MeOSuc-Ala-Ala-Pro-Val-p-nitroanilide is a PR3 substrate as well (13), PR3 is most likely present in GCF.…”

Elafin is specifically inactivated by RgpB from Porphyromonas gingivalis by distinct proteolytic cleavage

“…Nevertheless, enhanced local expression of elafin in the inflamed connective tissue of the periodontium is apparently not sufficient to control neutrophil proteases and gingival crevicular fluid contains the free elastase activity. Significantly, the level of elastase activity was found to correlate not only with the P. gingivalis infection but also with the presence of Rgps in gingival crevicular fluid (Mailhot et al, 1998). Therefore, our goal was to investigate whether gingipains, in comparison to proteases from Staphylococcus aureus , can interfere with the inhibitory activity of elafin and elucidate a mechanism of inhibitor interaction with gingipains.…”

Miropin, a Novel Bacterial Serpin from the Periodontopathogen Tannerella forsythia, Inhibits a Broad Range of Proteases by Using Different Peptide Bonds within the Reactive Center Loop

“…This inflammatory exudate contains high levels of neutrophil-derived serine proteases (e.g. neutrophil elastase and cathepsin G) (23). By degrading bacterial proteins, these proteases play an important role in the innate immune system of the host (24).…”