Utility of active MMP-8 as a test biomarker in periodontal and peri-implant diseases : diagnosis, prevention and treatment outcomes

Active Matrix Metalloproteinase (aMMP-8), collagenase-2, is a key mediator in the destruction of tissues seen in periodontitis and peri-implantitis. It can be detected in oral fluid samples, including gingival crevicular fluid (GCF), saliva, mouth rinse and peri-implant sulcular fluid (PISF). Active MMP-8, which is mainly expressed by neutrophils, can degrade almost all extracellular matrix and basement membrane components; it is causes particularly high levels of degradation in type I collagen, which is present in periodontal ligaments. In the first part of this thesis, we demonstrated that during an experimental gingivitis study aMMP-8 was not detectable in saliva. Therefore, this study confirms the evidence; aMMP-8 is detected in periodontitis. Simultaneously, the lateral-flow chairside aMMP-8 immunotests, PerioSafe and ImplantSafe with the Oralyzer reader, which are based on Professor Timo Sorsa’s research, were introduced and validated to analyse the levels of aMMP-8 in mouth rinse or PISF. Thus, the last parts of the thesis evaluate the accuracy and reliability of these developed tools: PerioSafe and ImplantSafe/Oralyzer, and whether they could provide an easy-to-use and quick way of identifying periodontitis and peri-implantitis. Our results indicated that the tools were successful in their detection. They were also very accurate in terms of both sensitivity and specificity with clinical periodontal parameters, such as bleeding on probing (BOP) and plaque index (PI). In addition, study V has investigated the effectiveness of sub-antimicrobial doses of doxycycline (SDD) treatment when combined with non-surgical periodontal therapy, and their effect on potential GCF biomarkers in periodontal tissue destruction over a period of twelve months. The analysed biomarkers were MMP-8, -9, -13, myeloperoxidase (MPO), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase 5 (TRAP 5), which were determined by various immunofluorometric analyses (IFMA) and enzyme-linked immunosorbent assays (ELISA). The results demonstrated reduced GCF levels in two MMPs that most commonly associated with periodontitis; MMP-8 and -9, and MPO, which potentially causes oxidative activation in them. However, MMP-13 and TRAP could be detected in the relevant GCF samples, whereas the OPG levels also decreased significantly as a result of adjunctive SDD therapy. This thesis showed the reliability of these tools, PerioSafe and ImplantSafe/Oralyzer, for the successful detection, prediction and monitoring of the progress of periodontitis and peri-implantitis, and their response to treatment. Additionally, we delved further in to the beneficial effects of sub-antimicrobial doses of doxycycline therapy.Active Matrix Metalloproteinase (aMMP-8), collagenase-2, is a key mediator in the destruction of tissues seen in periodontitis and peri-implantitis. It can be detected in oral fluid samples, including gingival crevicular fluid (GCF), saliva, mouth rinse and peri-implant sulcular fluid (PISF). Active MMP-8, which is mainly expressed by neutrophils, can degrade almost all extracellular matrix and basement membrane components; it is causes particularly high levels of degradation in type I collagen, which is present in periodontal ligaments. In the first part of this thesis, we demonstrated that during an experimental gingivitis study aMMP-8 was not detectable in saliva. Therefore, this study confirms the evidence; aMMP-8 is detected in periodontitis. Simultaneously, the lateral-flow chairside aMMP-8 immunotests, PerioSafe and ImplantSafe with the Oralyzer reader, which are based on Professor Timo Sorsa’s research, were introduced and validated to analyse the levels of aMMP-8 in mouth rinse or PISF. Thus, the last parts of the thesis evaluate the accuracy and reliability of these developed tools: PerioSafe and ImplantSafe/Oralyzer, and whether they could provide an easy-to-use and quick way of identifying periodontitis and peri-implantitis. Our results indicated that the tools were successful in their detection. They were also very accurate in terms of both sensitivity and specificity with clinical periodontal parameters, such as bleeding on probing (BOP) and plaque index (PI). In addition, study V has investigated the effectiveness of sub-antimicrobial doses of doxycycline (SDD) treatment when combined with non-surgical periodontal therapy, and their effect on potential GCF biomarkers in periodontal tissue destruction over a period of twelve months. The analysed biomarkers were MMP-8, -9, -13, myeloperoxidase (MPO), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase 5 (TRAP 5), which were determined by various immunofluorometric analyses (IFMA) and enzyme-linked immunosorbent assays (ELISA). The results demonstrated reduced GCF levels in two MMPs that most commonly associated with periodontitis; MMP-8 and -9, and MPO, which potentially causes oxidative activation in them. However, MMP-13 and TRAP could be detected in the relevant GCF samples, whereas the OPG levels also decreased significantly as a result of adjunctive SDD therapy. This thesis showed the reliability of these tools, PerioSafe and ImplantSafe/Oralyzer, for the successful detection, prediction and monitoring of the progress of periodontitis and peri-implantitis, and their response to treatment. Additionally, we delved further in to the beneficial effects of sub-antimicrobial doses of doxycycline therapy

Adjunctive Effects of a Sub-Antimicrobial Dose of Doxycycline on Clinical Parameters and Potential Biomarkers of Periodontal Tissue Catabolism

Objectives: The aim of the present randomized, double-blind, placebo-controlled, parallel-arm study was to examine the effectiveness of a sub-antimicrobial dose of doxycycline (SDD) in combination with nonsurgical periodontal therapy, compared to nonsurgical periodontal therapy alone, on potential gingival crevicular fluid (GCF) biomarkers of periodontal tissue catabolism related to the clinical outcomes over a 12-month period. Materials and Methods: GCF was collected and clinical parameters were recorded from 30 periodontitis patients randomized either to an SDD or placebo group. The SDD group received SDD (20 mg) b.i.d for 3 months plus scaling and root planing (SRP), while the placebo group was given placebo capsules b.i.d for 3 months plus SRP. The patients were evaluated every 3 months during the 12-month study period. At each visit, clinical parameters and GCF sampling were repeated. Matrix metalloproteinase (MMP)-8, MMP-9, MMP-13, myeloperoxidase (MPO), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase-5 (TRAP-5) were determined by IFMA and ELISA. Results: Significant improvements were observed in all clinical parameters in both groups over 12 months (p < 0.0125) while the SDD group showed significantly better reduction in gingival index (GI) and pocket depth and a gain in clinical attachment compared to the placebo group (p < 0.05). GCF MMP-8 and OPG levels significantly reduced in the SDD group compared to baseline (p < 0.05). GCF MMP-9 significantly decreased in both groups compared to baseline (p < 0.05). GCF MPO significantly decreased at 3 and 9 months in the SDD group, while it significantly decreased at 6 months in the placebo group (p < 0.05). TRAP and MMP-13 could be detected in none of the samples. Conclusions: The present results indicate that three months of adjunctive usage of SDD to nonsurgical periodontal therapy compared to nonsurgical periodontal therapy alone in periodontitis patients results in further improvement of clinical periodontal parameters and GCF markers of periodontal tissue breakdown over a 12-month period. Beneficial effects of adjunctive SDD therapy is likely to be related to the reduced levels of two major periodontitis-associated MMPs, MMP-8 and -9, and their potential oxidative activator MPO

Adjunctive Effects of a Sub-Antimicrobial Dose of Doxycycline on Clinical Parameters and Potential Biomarkers of Periodontal Tissue Catabolism

Objectives: The aim of the present randomized, double-blind, placebo-controlled, parallel-arm study was to examine the effectiveness of a sub-antimicrobial dose of doxycycline (SDD) in combination with nonsurgical periodontal therapy, compared to nonsurgical periodontal therapy alone, on potential gingival crevicular fluid (GCF) biomarkers of periodontal tissue catabolism related to the clinical outcomes over a 12-month period. Materials and Methods: GCF was collected and clinical parameters were recorded from 30 periodontitis patients randomized either to an SDD or placebo group. The SDD group received SDD (20 mg) b.i.d for 3 months plus scaling and root planing (SRP), while the placebo group was given placebo capsules b.i.d for 3 months plus SRP. The patients were evaluated every 3 months during the 12-month study period. At each visit, clinical parameters and GCF sampling were repeated. Matrix metalloproteinase (MMP)-8, MMP-9, MMP-13, myeloperoxidase (MPO), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase-5 (TRAP-5) were determined by IFMA and ELISA. Results: Significant improvements were observed in all clinical parameters in both groups over 12 months (p < 0.0125) while the SDD group showed significantly better reduction in gingival index (GI) and pocket depth and a gain in clinical attachment compared to the placebo group (p < 0.05). GCF MMP-8 and OPG levels significantly reduced in the SDD group compared to baseline (p < 0.05). GCF MMP-9 significantly decreased in both groups compared to baseline (p < 0.05). GCF MPO significantly decreased at 3 and 9 months in the SDD group, while it significantly decreased at 6 months in the placebo group (p < 0.05). TRAP and MMP-13 could be detected in none of the samples. Conclusions: The present results indicate that three months of adjunctive usage of SDD to nonsurgical periodontal therapy compared to nonsurgical periodontal therapy alone in periodontitis patients results in further improvement of clinical periodontal parameters and GCF markers of periodontal tissue breakdown over a 12-month period. Beneficial effects of adjunctive SDD therapy is likely to be related to the reduced levels of two major periodontitis-associated MMPs, MMP-8 and -9, and their potential oxidative activator MPO

Active MMP-8 (aMMP-8) as a Grading and Staging Biomarker in the Periodontitis Classification

The aim of this study was to investigate the utility of incorporating active matrix metalloproteinase-8 (aMMP-8) as a biomarker into the new periodontitis classification system (stage/grade) presented in 2018. This study included 150 Greek adults aged 25–78, of whom 74 were men and 76 women. Participants were tested with an aMMP-8 point-of-care mouthrinse test, after which a full-mouth clinical examination was performed to assess their periodontal and oral health. The aMMP-8 levels in mouthrinse were significantly lower among healthy patients compared with patients in more severe periodontitis stages and grades (Kruskal–Wallis test and Dunn–Bonferroni test for pairwise post-hoc comparisons; p < 0.01 and p < 0.05, respectively). Furthermore, aMMP-8 levels were less correlated with plaque levels than bleeding on probing (BOP) (Spearman’s rho = 0.269, p < 0.001; Spearman’s rho = 0.586, p < 0.001); respectively). Thus, aMMP-8 was more robust to the confounding effects of oral hygiene than traditional periodontal parameter bleeding on probing. The aMMP-8 point-of-care mouthrinse test can be utilized as an adjunctive and preventive diagnostic tool to identify periodontal disease, classified by stage and grade, and ongoing periodontal breakdown chairside in clinical practice in only 5 min. Overall, integrating aMMP-8 into the new periodontitis classification system seems beneficial

The Potential Role of Matrix Metalloproteinases 8 and 9 and Myeloperoxidase in Predicting Outcomes of Bacterial Meningitis of Childhood

Background. Matrix metalloproteinases (MMPs) and myeloperoxidase (MPO) contribute to the inflammatory cascade in the cerebrospinal fluid (CSF) during bacterial meningitis. We determined levels of MPO, MMP-8, MMP-9, and tissue inhibitor of metalloproteinase- (TIMP-) 1 in the CSF of children with bacterial meningitis and investigated how these inflammatory mediators relate to each other and to the disease outcomes. Methods. Clinical data and the diagnostic CSF samples from 245 children (median age eight months) with bacterial meningitis were obtained from a clinical trial in Latin America in 1996-2003. MMP-9 levels in the CSF were assessed by zymography, while MMP-8, MPO, and TIMP-1 concentrations were determined with immunofluorometric and enzyme-linked immunosorbent assays. Results. MPO correlated positively with MMP-8 (rho 0.496, PPeer reviewe

Active MMP-8 point-of-care (PoC)/chairside enzyme-test as an adjunctive tool for early and real-time diagnosis of peri-implantitis

Objective: The aim of this study was to investigate the utility of the active matrix metalloproteinase (aMMP-8)-point-of-care (PoC) test as a quantitative real-time chair-side diagnostic tool for peri-implant diagnosis, as well as assess the potentially developing and ongoing risk relative to the traditional clinical methods. Background: Current peri-implant and periodontal disease diagnoses rely on clinical arid radiological examinations. This case-control study investigated the applicability of aMMP-8-PoC immunotest for quantitative real-time diagnosis and monitoring of dental implants in health and disease. Methods: Sixty-eight patients visiting a specialist clinic for maintenance following dental implant placement underwent assessment of their peri-implant health. aMMP-8-PoC peri-implant sulcular fluid (PISF) lateral-flow immunotests were performed using ImplantSafe (R) technology quantitated by ORALyzer (R). In addition, the PISF samples were analyzed for total MMP-8, calprotectin, and interleukin (IL)-6 by enzyme-linked immunosorbent assays (ELISA), aMMP-8 by western immunoblot, and MMP-2 and MMP-9 by gelatin zymography. Results: The aMMP-8-PoC test promptly recorded and reflected peri-implant disease, differentiating it clearly from health. X-ray findings (bone loss > 2 mm), peri-implant pocket depth >= 3 mm, and bleeding on probing were significantly more prevalent among implants positive for the aMMP-8-PoC test. aMMP-8/ORALyzer analysis was more precise in recording disease than total MMP-8, calprotectin, IL-6, MMP-2, and MMP-9. Conclusions: The aMMP-8-PoC test can be conveniently implemented to alert for and detect active collagenolysis affecting peri-implant tissues, both in the early and advanced stages of the disease. Active and fragmented MMP-8 exhibits a strong and significant association with peri-implantitis as compared to total MMP-8 arid other biomarkers and can be utilized as the POC/chairside biomarker of choice in the new classification of peri-implantitis.Peer reviewe

The Ability of Quantitative, Specific, and Sensitive Point-of-Care/Chair-Side Oral Fluid Immunotests for aMMP-8 to Detect Periodontal and Peri-Implant Diseases

The analysis of the disease-specific oral and systemic biomarkers in saliva and oral fluids (i.e., mouth rinse, gingival crevicular fluid (GCF), and peri-implantitis fluid (PISF)) is demanding. Several hosts and microbial factors may influence their expression, release, and levels. The type of saliva/oral fluids utilized for the diagnostics affects the analysis. High sensitivity and specificities together with sophisticated methods and techniques are essential for valuable outcome. We describe here recently developed practical, convenient, inexpensive, noninvasive, and quantitative mouth rinse and PISF/GCF/chair-side/point-of-care (PoC) lateral-flow aMMP-8 immunoassays (PerioSafe and ImplantSafe/ORALyser) to detect, predict, and monitor successfully the course, treatment, and prevention of periodontitis and peri-implantitis, respectively. The tests have been independently and successfully validated to differentiate periodontal and peri-implant health and disease in Finland, Germany, Netherland, Sweden, Turkey, Nigeria, Malawi, and USA. The clinical use of salivary/oral fluid biomarkers to identify oral and systemic conditions requires additional studies utilizing these noninvasive screening, diagnostic, and preventive aMMP-8 PoC/chair-side technologies.Peer reviewe

Matrix Metalloproteinase-8 as an Inflammatory and Prevention Biomarker in Periodontal and Peri-Implant Diseases

Levels of and especially the degree of activation of matrix metalloproteinase (MMP-8) in oral fluids (i.e., saliva, mouth rinse, gingival crevicular fluid (GCF) and peri-implantitis sulcular fluid (PISF)) increase to pathologically elevated levels in the periodontal and peri-implant diseases. This study aimed at collecting and collating data from previously published studies and determining whether active MMP-8 (aMMP-8) could serve as a biomarker for the diagnosis and prevention of periodontal and peri-implant diseases. The literature search identified a total of 284 articles. Out of 284 articles, 61 articles were found to be relevant. Data obtained from the selected studies were combined, and it indicated that aMMP-8 in oral fluids exerts the strong potential to serve as a useful adjunctive diagnostic and preventive biotechnological tool in periodontal and peri-implant diseases. aMMP-8 can be used alone or in combination with other proinflammatory and/or microbiological biomarkers.Peer reviewe