Correlation Between Gingivain/Gingipain and Bacterial Dipeptidyl Peptidase Activity in Gingival Crevicular Fluid and Periodontal Attachment Loss in Chronic Periodontitis Patients. A 2‐Year Longitudinal Study

Abstract

The aim of this study is to determine whether either gingival crevicular fluid (GCF) bacterial gingivain/gingipain or dipeptidyl peptidase (DPP) levels, total activity (TA) and concentration (EC), predict progressive attachment loss (AL) in 75 patients with moderate periodontitis. GCF was collected from 16 molar and premolar mesiobuccal sites and then clinical attachment level (CAL) and probing depth (PD) were measured with an electronic constant pressure probe. Lastly, gingival, gingival bleeding, and plaque indices were scored. Prior to the baseline visit, patients were given basic periodontal treatment after which the above procedures were repeated. In addition, carefully localized radiographs were taken of the test teeth and repeated annually. Patients were then seen every 3 months for 2 years and the clinical measurements repeated at each visit. In 48 patients, 124 AL sites, 91 rapid AL (RAL), and 33 gradual AL (GAL) were detected. Gingivain/gingipain and bacterial DPP levels (TA and EC) at RAL sites were significantly higher (P ≤ 0.0001) than at paired control sites at the attachment loss time (ALT) and prediction time (PT). Mean levels over the study period of both proteases (TA and EC) at GAL sites were significantly higher (P ≤ 0.0001) than those at paired control sites. The GCF levels of gingivain/gingipain were always higher than those of DPP. Critical values (CV) of 5 μU/30 seconds (TA) and 30 μU/μL (EC) for both proteases showed high sensitivity and specificity values for TA and EC, which were the same at both ALT and PT. The positive predictive values were higher for gingivain/gingipain. Mean site levels, over the course of the study, of both proteases (TA and EC) were significantly higher (P ≥ 0.0001) at AL, RAL, and GAL sites than nonattachment loss (NAL) sites in AL patients and mean patient levels were significantly higher (P ≤ 0.0001) in AL, RAL, and GAL patients than NAL patients. These results indicate that both of these bacterial proteases in GCF may be predictors of periodontal attachment loss. J Periodontol 1996;67:703–716.

Functional Implication of the Hydrolysis of Platelet Endothelial Cell Adhesion Molecule 1 (CD31) by Gingipains ofPorphyromonas gingivalisfor the Pathology of Periodontal Disease

“…The present report indicates that gingipains at a concentration of 70 nM effectively reduce PECAM-1 expression on HUVECs in the presence of 10% serum, whereas limited hydrolysis of PECAM-1 was observed in the presence of 20% serum. The level of gingipains in crevicular fluid at inflamed sites was estimated at 40 to 90 nM (21), comparable to the amount used in this study. Hence, it is possible that proteolytic cleavage of PECAM-1 by gingipains occurs in vivo.…”

Gingipains from Porphyromonas gingivalis W83 Synergistically Disrupt Endothelial Cell Adhesion and Can Induce Caspase-Independent Apoptosis

“…Gingipain activity was calculated based on the average of three measurements of the maximum velocity of the enzymatic reaction of specific substrate turnover. The concentration of gingipain activity used was within reported physiological levels of gingipain activity found in periodontal lesions (17).…”

Biphasic Effect of Gingipains from Porphyromonas gingivalis on the Human Complement System

“…One strategy of the defense depends on the production of large amount of gingipains, which are cysteine proteases able to degrade, among other proteins, immunoglobulins and complement factors. Gingipains were found at the level of 100 nM in the gingival crevicular fluid collected from the inflamed sites of adult periodontitis patients (22). Furthermore, it is likely that, in situ in dental plaque heavily populated with P. gingivalis, the gingipain concentration is considerably higher than in the crevicular fluid because the majority of strains retain significant amounts of gingipains on the cell surface as well as release them in the form associated with vesicles (9).…”

“…Arginine-specific (HRgpA and RgpB) and lysine-specific (Kgp) gingipains were obtained from the P. gingivalis HG66 strain culture fluid as described previously (21,22). Briefly, Kgp and HRgpA were purified using gel filtration and affinity chromatography on arginine-Sepharose, whereas RgpB was purified using a combination of gel filtration and anion-exchange chromatography on Mono Q (GE Healthcare).…”