Characteristics of‐neutral proteases present in inflamed human gingiva

Abstract

– The existing forms of neutral proteases present in inflamed human gingiva were examined. Neutral 2 M KCl extracts of inflamed human gingival tissue were fractionated by gel filtration on Sephacryl S‐200 and the fractions were assayed for collagenase, trypsin‐, chymotrypsin‐, and elastase‐like proteases. Apparent molecular weights of 80–85 kDa were obtained for trypsin‐, chymotrypsin‐, and elastase‐like proteases, and 70–75 kDa for latent collagenase. Further fractionation of high molecular weight proteases on Con A‐Sepharose revealed that, unlike collagenase, chymotrypsin‐, and elastase‐like proteases, the trypsin‐like protease was bound by the affinity column. Native human placental type IV (basement membrane) collagen was degraded by chymotrypsin‐like and elastase‐like proteases but not by the trypsin‐like protease. This degradation was inhibited by phenylmethyl sulfonyl fluoride and EDTA. The serine proteases also degraded efficiently denatured type I collagen. No correlation of the activities of trypsin‐like protease and the other proteolytic enzymes was found in extracts of 18 individual gingival specimens. Significant correlation, however, was noted between collagenase and gelatinase. The gingival culture studies showed that, while the highest activity of the trypsin‐, chymotrypsin‐, and elastase‐like enzymes were measured in medium during first days of the culture, collagenase and gelatinase activities increased up to the fourth day of culture and stayed high until the end of the culture. These results suggest that the neutral proteases that may participate in the periodontal tissue destruction are produced by different cell types of gingiva.

Connective Tissue Degradation in Health and Periodontal Disease and the Roles of Matrix Metalloproteinases and Their Natural Inhibitors

“…In support of our original hypothesis are the findings that collagenase activity can be found in the crevicular fluid of patients with periodontitis in much larger amounts than in control subjects (Larivee et al, 1986;Villela et al, 1987). This collagenase seems to be only partly tissue-derived and mostly from PMNs (Villela et al, 1987;Suomalainen et al, 1989;Gangbar et al, 1990;Sorsa et al, 1990;Lee ef a/., 1991). TIMP levels could be measured only in healthy individuals or in clinically healthy sites (Larivee et al, 1986).…”

Lipid peroxidation: a possible role in the induction and progression of chronic periodontitis

“…1). The saliva lipid peroxidation concentration is (1) 167.55 ± 150.86 a LPO (pmol/site) (2) 93.02 ± 81.71 b GCF (ll) (3) 0.62 ± 0.28 c Healthy sites 66 LPO (lM) (1) 53.71 ± 92.70 a LPO (pmol/site) (2) 8.47 ± 15.90 b GCF (ll) (3) 0.13 ± 0.08 c correlated with the gingival crevicular fluid lipid peroxidation total amount (r ¼ 0.6378, p < 0.005).…”

Collagen Fibers and Inflammatory Cells in Healthy and Diseased Human Gingival Tissues: A Comparative and Quantitative Study by Immunohistochemistry and Automated Image Analysis

“…During periodontal diseases, the different types of gingival cells functioning as resident or inflammatory cells produce and release proteases (such as metalloproteinases) and cytokines which, together, generate periodontal tissue destruction. [4][5][6][7] Furthermore, inflammatory cells contain destructive enzymes that are able to degrade gingival tissue components if released during their functional activity or when the cells degenerate or die. 8 Many data also demonstrate the significant importance of the inflammatory mediators such as the cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in connective tissue degradation.…”