Molecular forms and fragments of salivary MMP‐8 in relation to periodontitis

Abstract

Aim To investigate the molecular forms of salivary matrix metalloproteinase (MMP)‐8 in relation to periodontitis. Materials and Methods Molecular forms, degree of activation and fragmentation of neutrophilic and mesenchymal‐type MMP‐8 isoforms were analysed from salivary samples of 81 subjects with generalized periodontitis, 63 subjects with localized periodontitis and 79 subjects without pocket teeth, by using western‐immunoblots with computer quantitation. In addition, human recombinant proMMP‐8 was in vitro activated by Treponema denticola chymotrypsin‐like protease (Td‐CTLP), sodium hypochlorite (NaOCl, 1 mM, oxidant) or amino phenyl mercuric acetate (APMA, 1 mM). Results In saliva of periodontitis‐affected individuals, MMP‐8 is found in multiple forms, that is, complexes, active and pro‐forms of neutrophilic and mesenchymal‐type MMP‐8, and especially 20–27 kDa fragments. The quantity of these fragments was elevated in both localized and generalized forms of periodontitis. Moreover, the tested activators (Td‐CTLP, NaOCl and APMA) activated inactive proMMP‐8, resulting in fragments of 20–27 kDa, in vitro, and salivary concentrations of T. denticola correlated significantly with salivary levels of fragmented MMP‐8. Conclusion The present results indicate that during the development and progression of periodontitis, MMP‐8 appears as activated and fragmented, and treponemal proteases most likely play role in this cascade.

Biological response to peri‐implantitis treatment

“…Studies have detected MMP‐8 in saliva and GCF of periodontitis patients and in PICF of PIP patients suggestive that MMP‐8 levels may be useful for diagnostic monitoring of the connective tissue destruction phase of peri‐implant disease. MMP‐8, including targeting its active form, has been reported to represent a valuable target for chairside point‐of‐care testing of oral fluids related to detection of periodontitis, and more recently in PIP; albeit active MMP‐8’s ability to affect treatment decision‐making remains to be fully elucidated and implemented within the global dental profession. Similar to our findings, Wang et al did not observe significant differences in the concentration of MMP‐8 between healthy and PIP individuals, although the literature would support that differences in selection of antibody systems to detect these types of biomolecules can create variation in the levels and outcome measures between health and disease .…”

Label‐Free Quantitative Proteomics versus Antibody‐Based Assays to Measure Neutrophil‐Derived Enzymes in Saliva

“…While various studies already demonstrated the presence of aMMP‐8 in periodontitis, the current study demonstrated for the first time that MMP‐8 is not activated during the induction and resolution of experimentally‐induced gingival inflammation, but rather exists in its latent proMMP‐8 form. We have used WB analysis to assess the molecular forms of MMP‐8 utilizing a polyclonal anti‐MMP‐8 antibody to specifically detect different MMP‐8 (complexes, pro and active forms, and smaller fragments) molecules by their molecular sizes and forms. For all saliva samples, the predominant molecular form identified by WB analysis was the full‐size pro‐form of MMP‐8, while no lower molecular size (20–55 kDa) MMP‐8 forms were detected throughout the study.…”

“…Since aMMP‐8, rather than the overall amount of MMP‐8, is strongly associated with the pathological destruction of connective tissue in periodontitis, the investigation of the degree of MMP‐8 activation, that is, conversion and fragmentation of latent 70–75 kDa proMMP‐8 to lower molecular size active and fragmented MMP‐8 20–55 kDa forms, is a very important and key issue . While various studies already demonstrated the presence of aMMP‐8 in periodontitis, the current study demonstrated for the first time that MMP‐8 is not activated during the induction and resolution of experimentally‐induced gingival inflammation, but rather exists in its latent proMMP‐8 form. We have used WB analysis to assess the molecular forms of MMP‐8 utilizing a polyclonal anti‐MMP‐8 antibody to specifically detect different MMP‐8 (complexes, pro and active forms, and smaller fragments) molecules by their molecular sizes and forms.…”

“…In periodontitis patients, compared to healthy individuals, salivary MMP‐8 was activated and fragmented . Since aMMP‐8, rather than the overall amount of MMP‐8, is strongly associated with the pathological destruction of connective tissue in periodontitis, the investigation of the degree of MMP‐8 activation, that is, conversion and fragmentation of latent 70–75 kDa proMMP‐8 to lower molecular size active and fragmented MMP‐8 20–55 kDa forms, is a very important and key issue . While various studies already demonstrated the presence of aMMP‐8 in periodontitis, the current study demonstrated for the first time that MMP‐8 is not activated during the induction and resolution of experimentally‐induced gingival inflammation, but rather exists in its latent proMMP‐8 form.…”

“…MMP‐8 is expressed primarily by neutrophils, and exists in various molecular forms, representing activity stages, including latent full‐size pro‐forms (proMMP‐8), active forms (aMMP‐8), higher molecular size complexes, and lower molecular size fragments . aMMP‐8 has been described as the major form present locally in patients with periodontitis . Assessing specifically aMMP‐8 enzymatic levels and degree of activation might support monitoring of disease progression.…”