Active MMP-8 Quantitative Test as an Adjunctive Tool for Early Diagnosis of Periodontitis

Marcela Hernández, Mauricio Baeza, Ismo T Räisänen, Johanna Contreras, Taina Tervahartiala, Alejandra Chaparro, Timo Sorsa, Patricia Hernández-Ríos, Marcela Hernández, Mauricio Baeza, Ismo T Räisänen, Johanna Contreras, Taina Tervahartiala, Alejandra Chaparro, Timo Sorsa, Patricia Hernández-Ríos

Abstract

Periodontitis is a host-mediated bacterial disease that affects the tooth attachment apparatus. Metalloproteinase-8 (MMP-8), a validated biomarker, could aid in clinical diagnosis. This study aimed to evaluate the diagnostic performance of active (a) MMP-8 immunotest versus total (t) MMP-8 ELISA for quantitative real-time diagnosis and assessment of periodontitis severity at the site level. Gingival crevicular fluid (GCF) was sampled from 30 healthy, 42 mild, and 59 severe periodontitis sites from thirty-one volunteers. MMP-8 concentrations were determined by time-resolved immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using the STATA package. Both active and total MMP-8-based methods discriminated among sites according to periodontal diagnosis and severity, with a positive correlation between the two tests (p < 0.001). (a) MMP-8 models showed the best performance in receiver operating characteristic (ROC) curves to discriminate between healthy and periodontitis sites (area under the curve [AUC] = 0.89), while (t) MMP-8 demonstrated a high diagnostic precision in the detection of mild from severe periodontitis sites (AUC ≥ 0.80). The use of (a) MMP-8 and (t) MMP-8 could represent a useful adjunctive tool for periodontitis diagnosis and severity. These results support the applicability of new point-of-care methods in the monitoring of high-risk periodontal patients.

Keywords: biomarkers; diagnosis; gingival crevicular fluid; matrix metalloproteinase-8; periodontal disease; periodontitis.

Conflict of interest statement

Timo Sorsa is the inventor of U.S. 5652,223, 5736,341, 5864,632, 6143,476 and US 2017/0023571A1 (issued 6 June 2019), WO 2018/060553 A1 (issued 31 May 2018), 10,488,415 B2, and US 2017/0023671A1 and Japanese Patent 2016-554676 and South-Korean patent 10-2016-7025378. The other authors report no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
MMP-8 levels in healthy and periodontitis sites. (A1) Active and (A2) total MMP-8 levels by diagnosis. (B1) Active and (B2) total MMP-8 levels by disease periodontitis severity. (a) active; (t) total. * p < 0.0001.
Figure 2
Figure 2
Diagnostic potential of MMP-8 tests to identify healthy sites. Diagnostic potential of active metalloproteinase-8 (a) MMP-8, to identify healthy from periodontitis sites, crude (A1) and adjusted (A2) models; diagnostic potential of (t) MMP-8 to identify healthy from periodontitis sites, crude (B1) and adjusted (B2) models; (C) sensitivity, specificity, and cut-off points of (a) MMP-8 and (t) MMP-8 diagnostic potential to identify healthy from periodontitis sites. AUC: area under the curve. CI: confidence interval. (a) MMP-8: active matrix metalloproteinase-8; (t) MMP-8: total matrix metalloproteinase-8. Immp8: IFMA MMP-8; Emmp8: ELISA MMP-8.
Figure 3
Figure 3
Diagnostic potential of biomarkers to identify severe periodontitis sites. Diagnostic potential of (a) MMP-8 to identify mild from severe periodontitis sites, crude (A1) and adjusted (A2) models; diagnostic potential of (t) MMP-8 to identify mild from severe periodontitis sites, crude (B1) and adjusted (B2) models; (C) sensitivity, specificity and cut-off points of (a) MMP-8 and (t) MMP-8. AUC: area under the curve. CI: confidence interval. (a) MMP-8: active matrix metalloproteinase-8; (t) MMP-8: total matrix metalloproteinase-8. Immp8: IFMA MMP-8; Emmp8: ELISA MMP-8.

References

    1. Baehni P., Tonetti M.S. On behalf of Group 1 of the European Workshop on Periodontology Conclusions and consensus statements on periodontal health, policy and education in Europe: A call for action-consensus view 1. Eur. J. Dent. Educ. 2010;14:2–3. doi: 10.1111/j.1600-0579.2010.00619.x.
    1. Tonetti M.S., Greenwell H., Kornman K.S. Staging and grading of periodontitis: Framework and proposal of a new classification and case definition. J. Periodontol. 2018;89:S159–S172. doi: 10.1002/JPER.18-0006.
    1. Buduneli N., Kinane D.F. Host-derived diagnostic markers related to soft tissue destruction and bone degradation in periodontitis. J. Clin. Periodontol. 2011;38:85–105. doi: 10.1111/j.1600-051X.2010.01670.x.
    1. Giannobile W. Crevicular fluid biomarkers of oral bone loss. Curr. Opin. Periodontol. 1997;4:11–20.
    1. Armitage G.C. Analysis of gingival crevice fluid and risk of progression of periodontitis. Periodontol. 2000. 2004;34:109–119. doi: 10.1046/j.0906-6713.2002.003427.x.
    1. Sorsa T., Gursoy U.K., Nwhator S., Hernández M., Tervahartiala T., Leppilahti J., Gürsoy M., Könönen E., Emingil G., Pussinen P., et al. Analysis of matrix metalloproteinases, especially MMP-8, in gingival crevicular fluid, mouthrinse and saliva for monitoring periodontal diseases. Periodontol. 2000. 2015;70:142–163. doi: 10.1111/prd.12101.
    1. Franco C., Patricia H.-R., Timo S., Claudia B., Marcela H. Matrix Metalloproteinases as Regulators of Periodontal Inflammation. Int. J. Mol. Sci. 2017;18:440. doi: 10.3390/ijms18020440.
    1. Golub L.M., Lee H.M., Greenwald R.A., Ryan M.E., Sorsa T., Salo T., Giannobile W. A matrix metalloproteinase inhibitor reduces bone-type collagen degradation fragments and specific collagenases in gingival crevicular fluid during adult periodontitis. Inflamm. Res. 1997;46:310–319. doi: 10.1007/s000110050193.
    1. Ingman T., Tervahartiala T., Ding Y., Tschesche H., Haerian A., Kinane D.F., Konttinen Y.T., Sorsa T. Matrix metalloproteinases and their inhibitors in gingival crevicular fluid and saliva of periodontitis patients. J. Clin. Periodontol. 1996;23:1127–1132. doi: 10.1111/j.1600-051X.1996.tb01814.x.
    1. Ingman T., Sorsa T., Suomalainen K., Halinen S., Lindy O., Lauhio A., Saari H., Konttinen Y.T., Golub L.M. Tetracycline Inhibition and the Cellular Source of Collagenase in Gingival Crevicular Fluid in Different Periodontal Diseases. A Review Article. J. Periodontol. 1993;64:82–88. doi: 10.1902/jop.1993.64.2.82.
    1. Hernández M., Baeza M., Contreras J., Sorsa T., Tervahartiala T., Valdés M., Chaparro A., Hernández-Ríos P. MMP-8, TRAP-5, and OPG Levels in GCF Diagnostic Potential to Discriminate between Healthy Patients’, Mild and Severe Periodontitis Sites. Biomolecules. 2020;10:1500. doi: 10.3390/biom10111500.
    1. Baeza M., Garrido M., Hernández-Ríos P., Dezerega A., García-Sesnich J., Strauss F., Aitken J.P., Lesaffre E., Vanbelle S., Gamonal J., et al. Diagnostic accuracy for apical and chronic periodontitis biomarkers in gingival crevicular fluid: An exploratory study. J. Clin. Periodontol. 2016;43:34–45. doi: 10.1111/jcpe.12479.
    1. Sorsa T., Alassiri S., Grigoriadis A., Räisänen I.T., Pärnänen P., Nwhator S.O., Gieselmann D.-R., Sakellari D. Active MMP-8 (aMMP-8) as a Grading and Staging Biomarker in the Periodontitis Classification. Diagnostics. 2020;10:61. doi: 10.3390/diagnostics10020061.
    1. Arias-Bujanda N., Regueira-Iglesias A., Balsa-Castro C., Nibali L., Donos N., Tomás I. Accuracy of single molecular biomarkers in gingival crevicular fluid for the diagnosis of periodontitis: A systematic review and meta-analysis. J. Clin. Periodontol. 2019;46:1166–1182. doi: 10.1111/jcpe.13188.
    1. Sorsa T., Tjäderhane L., Konttinen Y.T., Lauhio A., Salo T., Lee H., Golub L.M., Brown D.L., Mäntylä P. Matrix metalloproteinases: Contribution to pathogenesis, diagnosis and treatment of periodontal inflammation. Ann. Med. 2006;38:306–321. doi: 10.1080/07853890600800103.
    1. Sorsa T., Hernández M., Leppilahti J., Munjal S., Netuschil L., Mäntylä P. Detection of gingival crevicular fluid MMP-8 levels with different laboratory and chair-side methods. Oral Dis. 2009;16:39–45. doi: 10.1111/j.1601-0825.2009.01603.x.
    1. Teles R., Sakellari D., Teles F., Konstantinidis A., Kent R., Socransky S., Haffajee A. Relationships Among Gingival Crevicular Fluid Biomarkers, Clinical Parameters of Periodontal Disease, and the Subgingival Microbiota. J. Periodontol. 2010;81:89–98. doi: 10.1902/jop.2009.090397.
    1. Hanemaaijer R., Sorsa T., Konttinen Y.T., Ding Y., Sutinen M., Visser H., van Hinsbergh V.W.M., Helaakoski T., Kainulainen T., Rönkä H., et al. Matrix Metalloproteinase-8 Is Expressed in Rheumatoid Synovial Fibroblasts and Endothelial Cells. J. Biol. Chem. 1997;272:31504–31509. doi: 10.1074/jbc.272.50.31504.
    1. Steen P.E.V.D., Wuyts A., Husson S.J., Proost P., Van Damme J., Opdenakker G. Gelatinase B/MMP-9 and neutrophil collagenase/MMP-8 process the chemokines human GCP-2/CXCL6, ENA-78/CXCL5 and mouse GCP-2/LIX and modulate their physiological activities. JBIC J. Biol. Inorg. Chem. 2003;270:3739–3749. doi: 10.1046/j.1432-1033.2003.03760.x.
    1. Hernández M., Martínez B., Tejerina J.M., Valenzuela M.A., Gamonal J. MMP-13 and TIMP-1 determinations in progressive chronic periodontitis. J. Clin. Periodontol. 2007;34:729–735. doi: 10.1111/j.1600-051X.2007.01107.x.
    1. Chapple I.L., Mealey B.L., Van Dyke T.E., Bartold P.M., Dommisch H., Eickholz P., Geisinger M.L., Genco R.J., Glogauer M., Goldstein M., et al. Periodontal health and gingival diseases and conditions on an intact and a reduced periodontium: Consensus report of workgroup 1 of the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. J. Periodontol. 2018;89:S74–S84. doi: 10.1002/JPER.17-0719.
    1. Ríos M.H., Sorsa T., Obregón F., Tervahartiala T., Valenzuela M.A., Pozo P., Dutzan N., Lesaffre E., Molas M., Gamonal J. Proteolytic roles of matrix metalloproteinase (MMP)-13 during progression of chronic periodontitis: Initial evidence for MMP-13/MMP-9 activation cascade. J. Clin. Periodontol. 2009;36:1011–1017. doi: 10.1111/j.1600-051X.2009.01488.x.
    1. Hemmilä I., Dakubu S., Mukkala V.-M., Siitari H., Lövgren T. Europium as a label in time-resolved immunofluorometric assays. Anal. Biochem. 1984;137:335–343. doi: 10.1016/0003-2697(84)90095-2.
    1. Arias-Bujanda N., Regueira-Iglesias A., Blanco-Pintos T., Alonso-Sampedro M., Relvas M., González-Peteiro M.M., Balsa-Castro C., Tomás I., Sampedro-Alonso M. Diagnostic accuracy of IL1β in saliva: The development of predictive models for estimating the probability of the occurrence of periodontitis in non-smokers and smokers. J. Clin. Periodontol. 2020;47:702–714. doi: 10.1111/jcpe.13285.
    1. Gursoy U.K., Könönen E., Pradhan-Palikhe P., Tervahartiala T., Pussinen P., Suominen-Taipale L., Sorsa T. Salivary MMP-8, TIMP-1, and ICTP as markers of advanced periodontitis. J. Clin. Periodontol. 2010;37:487–493. doi: 10.1111/j.1600-051X.2010.01563.x.
    1. Romanelli R., Mancini S., Laschinger C., Overall C.M., Sodek J., McCulloch C.A.G. Activation of Neutrophil Collagenase in Periodontitis. Infect. Immun. 1999;67:2319–2326. doi: 10.1128/IAI.67.5.2319-2326.1999.
    1. Kiili M., Cox S.W., Chen H.W., Wahlgren J., Maisi P., Eley B.M., Salo T., Sorsa T. Collagenase-2 (MMP-8) and collagenase-3 (MMP-13) in adult periodontitis: Molecular forms and levels in gingival crevicular fluid and immunolocalisation in gingival tissue. J. Clin. Periodontol. 2002;29:224–232. doi: 10.1034/j.1600-051x.2002.290308.x.
    1. Lee W., Aitken S., Sodek J., McCulloch C.A.G. Evidence of a direct relationship between neutrophil collagenase activity and periodontal tissue destruction in vivo: Role of active enzyme in human periodontitis. J. Periodontal Res. 1995;30:23–33. doi: 10.1111/j.1600-0765.1995.tb01249.x.
    1. Mancini S., Romanelli R., Laschinger C.A., Overall C.M., Sodek J., McCulloch C.A. Assessment of a Novel Screening Test for Neutrophil Collagenase Activity in the Diagnosis of Periodontal Diseases. J. Periodontol. 1999;70:1292–1302. doi: 10.1902/jop.1999.70.11.1292.
    1. Räisänen I.T., Lähteenmäki H., Gupta S., Grigoriadis A., Sahni V., Suojanen J., Seppänen H., Tervahartiala T., Sakellari D., Sorsa T. An aMMP-8 Point-of-Care and Questionnaire Based Real-Time Diagnostic Toolkit for Medical Practitioners. Diagnostics. 2021;11:711. doi: 10.3390/diagnostics11040711.
    1. Deng K., Pelekos G., Jin L., Tonetti M.S. Diagnostic accuracy of a point-of-care aMMP-8 test in the discrimination of periodontal health and disease. J. Clin. Periodontol. 2021;48:1051–1065. doi: 10.1111/jcpe.13485.
    1. Leppilahti J.M., Hernández-Ríos P.A., Gamonal J.A., Tervahartiala T., Brignardello-Petersen R., Mäntylä P., Sorsa T., Hernández M. Matrix metalloproteinases and myeloperoxidase in gingival crevicular fluid provide site-specific diagnostic value for chronic periodontitis. J. Clin. Periodontol. 2013;41:348–356. doi: 10.1111/jcpe.12223.
    1. Mäntylä P., Stenman M., Kinane D.F., Tikanoja S., Luoto H., Salo T., Sorsa T. Gingival crevicular fluid collagenase-2 (MMP-8) test stick for chair-side monitoring of periodontitis. J. Periodontal Res. 2003;38:436–439. doi: 10.1034/j.1600-0765.2003.00677.x.
    1. Ramseier C.A., Kinney J.S., Herr A., Braun T., Sugai J.V., Shelburne C.A., Rayburn L.A., Tran H.M., Singh A.K., Giannobile W.V. Identification of Pathogen and Host-Response Markers Correlated With Periodontal Disease. J. Periodontol. 2009;80:436–446. doi: 10.1902/jop.2009.080480.
    1. Salminen A., Gursoy U.K., Paju S., Hyvärinen K., Mäntylä P., Buhlin K., Könönen E., Nieminen M.S., Sorsa T., Sinisalo J., et al. Salivary biomarkers of bacterial burden, inflammatory response, and tissue destruction in periodontitis. J. Clin. Periodontol. 2014;41:442–450. doi: 10.1111/jcpe.12234.
    1. Leppilahti J.M., Kallio M.A., Tervahartiala T., Sorsa T., Mäntylä P. Gingival Crevicular Fluid Matrix Metalloproteinase-8 Levels Predict Treatment Outcome Among Smokers With Chronic Periodontitis. J. Periodontol. 2014;85:250–260. doi: 10.1902/jop.2013.130156.
    1. Lorenz K., Keller T., Noack B., Freitag A., Netuschil L., Hoffmann T. Evaluation of a novel point-of-care test for active matrix metalloproteinase-8: Agreement between qualitative and quantitative measurements and relation to periodontal inflammation. J. Periodontal Res. 2016;52:277–284. doi: 10.1111/jre.12392.
    1. Sorsa T., Mäntylä P., Ronka H., Kallio P., Kallis G.-B., Lundqvist C., Kinane D.F., Salo T., Golub L.M., Teronen O., et al. Scientific Basis of a Matrix Metalloproteinase-8 Specific Chair-side Test for Monitoring Periodontal and Peri-implant Health and Disease. Ann. N. Y. Acad. Sci. 1999;878:130–140. doi: 10.1111/j.1749-6632.1999.tb07679.x.
    1. Al-Majid A., Alassiri S., Rathnayake N., Tervahartiala T., Gieselmann D.-R., Sorsa T. Matrix Metalloproteinase-8 as an Inflammatory and Prevention Biomarker in Periodontal and Peri-Implant Diseases. Int. J. Dent. 2018;2018:1–27. doi: 10.1155/2018/7891323.
    1. Dejonckheere E., Vandenbroucke R., Libert C. Matrix metalloproteinase8 has a central role in inflammatory disorders and cancer progression. Cytokine Growth Factor Rev. 2011;22:73–81. doi: 10.1016/j.cytogfr.2011.02.002.

Source: PubMed

Sponsorit ja yhteistyökumppanit

Sairaudet

Huumeiden interventiot

3
Tilaa