Identification of proteases from periodontopathogenic bacteria as activators of latent human neutrophil and fibroblast-type interstitial collagenases

Abstract

Activation of latent human fibroblast-type and neutrophil interstitial procollagenases as well as degradation of native type I collagen by supraand subgingival dental plaque extracts, an 80-kDa trypsinlike protease from Porphyromas gingivalis (ATCC 33277), a 95-kDa chymotrypsinlike protease from Treponema denticola (ATCC 29522), and selected bacterial species commonly isolated in periodontitis was studied. The bacteria included were PrevotelUa intermedia (ATCC 25261), PrevoteUla buccae (ES 57), PrevoteUa oris (ATCC 33573), Porphyromonas endodontalis (ES 54b), Actinobacillus actinomycetemcomitans (ATCC 295222), Fusobacterium nukeatum (ATCC 10953), Mitsuokela dentalis (DSM 3688), and Streptococcus mitis (ATCC 15909). None of the bacteria activated latent procollagenases; however, both sub-and supragingival dental plaque extracts (neutral salt extraction) and proteases isolated from cell extracts from potentially periodontopathogenic bacteria P. gingivalis and T. denticola were found to activate latent human fibroblast-type and neutrophil interstitial procollagenases. The fibroblast-type interstitial collagenase was more efficiently activated by bacterial proteases than the neutrophil counterpart, which instead preferred nonproteolytic activation by the oxidative agent hypochlorous acid. The proteases were not able to convert collagenase tissue inhibitor of metalloproteinase (TIMP-1) complexes into active form or to change the ability of TIMP-1 to inhibit interstitial collagenase. None of the studied bacteria, proteases from P. gingivalis and T. denticola, or extracts of supraand subgingival dental plaque showed any significant collagenolytic activity. However, the proteases degraded native and denatured collagen fragments after cleavage by interstitial collagenase and gelatinase. Our results indicate that proteases from periodontopathogenic bacteria can act as direct proteolytic activators of human procollagenases and degrade collagen fragments. Thus, in concert with host enzymes the bacterial proteases may participate in periodontal tissue destruction.

Cellular source, activation and inhibition of dental plaque collagenase

“…We have previously addressed the cel!ular sources and activation of collagenases (MMP-1 and -8), gelatinases/type IV collagenases (MMP-2 and -9) as well as serine proteinases elastase and cathepsin G in gingival tissue, GCF and saliva of healthy and diabetic aduU periodontitis patients, with localized juvenile periodontitis as well as patients 'with osseointegrated dental implants (Sorsa et al 1988, Sorsa 1989, Sorsa et al, 1990a, b, Uitto et al, 1990, Suomalainen et al 1991a, b, Suomalainen 1993, Ingman et al 1993a, 1994a, b, c, Tervahartiala et al 1995. We have also found significant differences in the sensitivities of human fibrobiast-type (MMP-1), neutrophil (MMP-8) interstitial collagenases and P. gingivalis collagenases to doxycycline-inhibition (Suomalainen et ai, 1992, Golub et al 1992, Goiub et al 1993, Ingman et al 1993b). These observations may have significant therapeutic and diagnostic imphcations (Golub et al 1992, 1993, Ingman et al, 1993b, Furthermore, recent studies have shown that certain proteases from potent periodontopathogenic bacteria can act as direct proteolytic activators of latent human proMMP-1 and -8 as well as induce their secretion by cultured gingival fibroblasts and oral keratinocytes , DeCarlo 1994.…”

Interfacial chemistry of moisture‐aged class II composite restorations

“…The exposed collagen could be attacked and degraded by bacterial enzymes. 17,46 The cumulative effect would be breakdown of the adhesive/dentin bond, and ultimately, undermining of the marginal integrity of the composite restoration.…”

The relationship between colonization and haemagglutination inhibiting and B cell epitopes of Porphyromonas gingivalis

“…The bacterium elaborates numerous potential virulence determinants, including proteases, which may provoke or deregulate the inflammatory response in the host tissues, which in turn can lead to destructive disease [4]. In addition, in vitro studies suggest that proteases may contribute directly to destruction of host tissue [5]. The gene encoding an extracellular arginine-specific protease of P. gingivalis has been cloned and sequenced [6,7].…”