Active MMP-8 Research Articles

Induction of Collagenolytic MMP-8 and -9 Tissue Destruction Cascade in Mouth by Head and Neck Cancer Radiotherapy: A Cohort Study.

The effect of head and neck cancer (HNC) radiotherapy (RT) on biomarkers is not known but there is a lot of potential for gaining more precise cancer treatments and less side effects. This cohort study investigated the levels and molecular forms of the matrix metalloproteinase (MMP) -8 and -9, tissue inhibitor of metalloproteinase (TIMP)-1, myeloperoxidase (MPO) and interleukin (IL)-6 in mouth-rinse samples as well as the clinical periodontal status in HNC patients (n = 21) receiving RT. Complete periodontal examinations were performed pre-RT and one month after RT. Mouth-rinse samples (pre-RT, after six weeks of RT and one month after RT) were assayed using a point-of-care-kit (PerioSafe®/ORALyzer® (Dentognostics GmbH, Jena, Germany)) for active MMP-8 and ELISA analysis for total MMP-8 and -9, MPO, TIMP-1, and IL-6 levels. Molecular forms of MMP-9 were assessed by gelatinolytic zymography and MMP-8 by western immunoblot. Significant changes were observed between the three time points in the mean levels of active and total MMP-8, active MMP-9, and IL-6. Their levels increased during the RT and decreased after the RT period. The aMMP-8 levels stayed elevated even one month after RT compared to the pre-RT. Clinical attachment loss, probing depths, and bleeding on probing were increased between pre- and post-calculations in periodontal status. Elevated inflammatory biomarker levels together with clinical recordings strongly suggest that RT eventually increases the risk to the periodontal tissue destruction by inducing the active proteolytical MMP-cascade, and especially by prolonged activity of collagenolytic aMMP-8. Eventually, the aMMP-8 point-of-care mouth-rinse test could be an easy, early detection tool for estimating the risk for periodontal damage by the destructive MMP-cascade in HNC patients with RT treatment.

Active MMP-8 as a Biomarker of Peri-implant Health or Disease.

This study investigated the potential of testing for active matrix metalloproteinase-8 (aMMP-8) by a quantitative point-of-care (PoC), chairside, lateral flow immunotest as a biomarker for the presence or absence of peri-implant diseases. Eighty healthy patients with implants were recruited. The samples were collected from peri-implant sulcular fluid and quantitatively analyzed for aMMP-8. Clinical indices, which included probing depth, clinical attachment loss, bleeding on probing, and plaque, were recorded and radiographic assessments were performed. Comparisons of aMMP-8 levels and clinical parameters were analyzed by the Kruskal-Wallis test and the pairwise post hoc Dunn-Bonferroni test. A receiver operating curve analysis was used to analyze the diagnostic ability of aMMP-8 and the correlation between aMMP-8 and probing depth of the sampled site was sought by Spearman's rho and the coefficient of determination (R 2). Statistical analysis revealed statistically significant differences of aMMP-8 levels between the healthy and the mucositis and peri-implantitis groups, and between the mucositis and the peri-implantitis groups. Increasing probing depths of the sampled site and aMMP-8 levels were significantly correlated. These data suggest that the aMMP-8 PoC test can be a beneficial adjunctive tool for early identification and screening of the risk of peri-implant diseases and progression.

Validation of a noninvasive aMMP-8 point-of-care diagnostic methodology in COVID-19 patients with periodontal disease.

ObjectivesThe aim of this study was to validate an active matrix metalloproteinase (MMP‐8) point‐of‐care diagnostic tool in COVID‐19 patients with periodontal disease.Subjects, Materials, and MethodsSeventy‐two COVID‐19‐positive and 30 COVID‐19‐negative subjects were enrolled in the study. Demographic data were recorded, periodontal examination carried out, and chairside tests run for evaluating the expression of active MMP‐8 (aMMP‐8) in the site with maximum periodontal breakdown via gingival crevicular fluid sampling as well as via a mouth rinse‐based kit for general disease activity. In COVID‐19‐positive patients, the kits were run again once the patients turned COVID‐19 negative.ResultsThe overall (n = 102) sensitivity/specificity of the mouthrinse‐based kits to detect periodontal disease was 79.41%/36.76% and that of site‐specific kits was 64.71%/55.88% while adjusting for age, gender, and smoking status increased the sensitivity and specificity (82.35%/76.47% and 73.53%/88.24, respectively). Receiver operating characteristic (ROC) analysis for the adjusted model revealed very good area under the ROC curve 0.746–0.869 (p < .001) and 0.740–0.872 (p < .001) (the aMMP‐8 mouth rinse and site‐specific kits, respectively). No statistically significant difference was observed in the distribution of results of aMMP‐8 mouth rinse test (p = .302) and aMMP‐8 site‐specific test (p = .189) once the subjects recovered from COVID‐19.ConclusionsThe findings of the present study support the aMMP‐8 point‐of‐care testing (PoCT) kits as screening tools for periodontitis in COVID‐19 patients. The overall screening accuracy can be further increased by utilizing adjunctively risk factors of periodontitis. The reported noninvasive, user‐friendly, and objective PoCT diagnostic methodology may provide a way of stratifying risk groups, deciding upon referrals, and in the institution of diligent oral hygiene regimens.

Soft tissue response to different abutment materials: A controlled and randomized human study using an experimental model.

Aim of this study was to compare the soft tissue response to implant abutments made of titanium, zirconia, zirconia veneered with feldspar ceramics and PEEK by various clinical, histological, microbiological, and molecular biological markers in an experimental model. A total of 40 experimental one-piece healing abutments of four different materials were mounted on bone level implants in 20 volunteering patients (split-mouth design). After a three-month period of open healing, clinical parameters at the abutments were assessed and adjacent mucosa was sampled for inflammatory cytokine mRNA concentrations and histological analysis by a novel method. In addition, PISF samples were obtained for the analysis of periodonto-pathogenic bacteria counts and active MMP-8 levels. Marginal bone level change was measured by intra oral radiographs. Abutments of the different materials did not exhibit significant differences regarding clinical parameters, pathogenic bacteria counts or pro-inflammatory cytokine concentrations. Likewise, no significant differences were detected regarding soft tissue morphology or bone level change. Compared to titanium abutments, significantly less mononuclear inflammatory cells were detected in the mucosa at abutments made of zirconia veneered with feldspar ceramics. All examined abutment materials exhibited a similar soft tissue response compared to titanium and histological data did not reveal early signs of elevated inflammation caused by PEEK- and feldspar-veneered zirconia abutments. Due to the short observation period and the small sample size, a final conclusion on the long-term suitability of those abutment materials cannot be drawn. However, based on the presented data, we consider further studies on that subject as appropriate.

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

ObjectiveThe 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.BackgroundCurrent peri‐implant and periodontal disease diagnoses rely on clinical and 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.MethodsSixty‐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® technology quantitated by ORALyzer®. 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.ResultsThe 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.ConclusionsThe 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 and other biomarkers and can be utilized as the POC/chairside biomarker of choice in the new classification of peri‐implantitis.

Linking oral microbial proteolysis to aMMP-8 PoC diagnostics along with the stage and grade of periodontitis: A cross-sectional study.

Linking oral microbial proteolysis to aMMP-8 PoC diagnostics along with the stage and grade of periodontitis: A cross-sectional study.

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

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.

Periodontitis and peri-implantitis tissue levels of Treponema denticola-CTLP and its MMP-8 activating ability

Background and AimsChymotrypsin-like-proteinase of Treponema denticola (Td-CTLP) can stimulate the protein expression and activation of matrix metalloproteinase (MMP)-8 (or collagenase-2), a potent tissue destructive enzyme from gingival cells in vitro. The aims of this study were 1) to demonstrate the proMMP-8 (or latent MMP-8) activation by Td-CTLP in vitro and 2) to detect Td-CTLP and MMP-8 protein levels in the tissue samples of peri-implantitis and periodontitis patients. Materials and MethodsproMMP-8 activation by Td-CTLP was analyzed by immunoblots. Tissue specimens were collected from 38 systemically healthy and non-smoking patients; 14 of whom had moderate to severe periodontitis, 10 of whom were suffering from peri-implantitis, and finally 14 of whom showed no sign of periodontal inflammation nor radiological bone decay (control group). The immune-expression levels of MMP-8 and Td-CTLP in the epithelium and the connective tissue were analyzed immunohistochemically. A pixel color-intensity analyze was performed with ImageJ software (version 1.46c; Rasband WS, National Institutes of Health, Bethesda, MD, USA) to obtain a comparable numeral score for each patient’s epithelium and connective tissue MMP-8 and Td-CTLP enzyme level. ResultsTd-CTLP activated proMMP-8 in vitro by converting the 70−75 kDa proMMP-8 to 65 kDa active MMP-8. Also, lower molecular size 25−50 kDa parts of MMP-8 were formed. There was no statistically significant difference between the study groups in terms of their MMP-8 and Td-CTLP levels in the epithelium or in the connective tissue. ConclusionRegarding the limits of this study, it can thus be said that the Td-CTLP enzyme can activate the host proMMP-8 enzyme. Tissue protein levels of MMP-8 and Td-CTLP do not seem to be changed in peri-implantitis and in periodontitis.

Expression of Matrix Metalloproteinases-8 and Myeloperoxidase in Pulp Tissue after Pulpotomy with Calcium Silicate Cements

Objective: To evaluate the pulp tissue of rat molars after pulpotomies with mineral trioxide aggregate (MTA), BiodentineTM (BDT) and calcium hydroxide (CH) mixed with sterile saline solution (24 hours, 72 hours, 7 days and 15 days), through correlating MPO activity with active neutrophils and MMP8 activity with tissue remodeling. Material and Methods: Thirty-eight Wistar rats were randomly distributed into groups (control, I (MTA gray), II (BDT), and III (CH)) and subdivided according to the study period of 24 hours, 72 hours, 7 days or 15 days after pulpotomy. MMP8 activity was assessed through fluorescence technique, and MPO activity was determined using the MPO assay. Results: A gradual decrease of MPO and MM8 activity occurred in the group MTA over the experimental periods (p<0.05). Groups BDT and CH exhibited an increase in the activity at 7 and 15 days (p<0.05). Conclusion: MTA demonstrated a decrease in the values of MPO e MMP8. BDT and CH showed high neutrophilic and collagenase activity over the experimental periods.

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.

Tissue Inhibitor of Metalloproteinase-1 Promotes Polymorphonuclear Neutrophil (PMN) Pericellular Proteolysis by Anchoring Matrix Metalloproteinase-8 and -9 to PMN Surfaces.

Matrix metalloproteinase (MMP)-8 and -9 released by degranulating polymorphonuclear cells (PMNs) promote pericellular proteolysis by binding to PMN surfaces in a catalytically active tissue inhibitor of metalloproteinases (TIMP)-resistant forms. The PMN receptor(s) to which MMP-8 and MMP-9 bind(s) is not known. Competitive binding experiments showed that Mmp-8 and Mmp-9 share binding sites on murine PMN surfaces. A novel form of TIMP-1 (an inhibitor of soluble MMPs) is rapidly expressed on PMN surfaces when human PMNs are activated. Membrane-bound TIMP-1 is the PMN receptor for pro- and active MMP-8 and -9 as shown by the following: 1) TIMP-1 is strikingly colocalized with MMP-8 and -9 on activated human PMN surfaces and in PMN extracellular traps; 2) minimal immunoreactive and active Mmp-8 or Mmp-9 are detected on the surface of activated Timp-1-/- murine PMNs; and 3) binding of exogenous Timp-1 (but not Timp-2) to Timp-1-/- murine PMNs reconstitutes the binding of exogenous pro-Mmp-8 and pro-Mmp-9 to the surface of Timp-1-/- PMNs. Unlike full-length pro-Mmp-8 and pro-Mmp-9, mutant pro-Mmp proteins lacking the COOH-terminal hemopexin domain fail to bind to Mmp-8-/-x Mmp-9-/- murine PMNs. Soluble hemopexin inhibits the binding of pro-Mmp-8 and pro-Mmp-9 to Mmp-8-/-x Mmp-9-/- murine PMNs. Thus, the COOH-terminal hemopexin domains of pro-Mmp-8 and pro-Mmp-9 are required for their binding to membrane-bound Timp-1 on murine PMNs. Exposing nonhuman primates to cigarette smoke upregulates colocalized expression of TIMP-1 with MMP-8 and MMP-9 on peripheral blood PMN surfaces. By anchoring MMP-8 and MMP-9 to PMN surfaces, membrane-bound TIMP-1 plays a counterintuitive role in promoting PMN pericellular proteolysis occurring in chronic obstructive pulmonary disease and other diseases.

Targeting MMP-9 in Diabetic Foot Ulcers.

Diabetic foot ulcers (DFUs) are significant complications of diabetes and an unmet medical need. Matrix metalloproteinases (MMPs) play important roles in the pathology of wounds and in the wound healing process. However, because of the challenge in distinguishing active MMPs from the two catalytically inactive forms of MMPs and the clinical failure of broad-spectrum MMP inhibitors in cancer, MMPs have not been a target for treatment of DFUs until recently. This review covers the discovery of active MMP-9 as the biochemical culprit in the recalcitrance of diabetic wounds to healing and targeting this proteinase as a novel approach for the treatment of DFUs. Active MMP-8 and MMP-9 were observed in mouse and human diabetic wounds using a batimastat affinity resin and proteomics. MMP-9 was shown to play a detrimental role in diabetic wound healing, whereas MMP-8 was beneficial. A new class of selective MMP-9 inhibitors shows clinical promise for the treatment of DFUs.

Active Matrix Metalloproteinase-8 Point-of-Care (PoC)/Chairside Mouthrinse Test vs. Bleeding on Probing in Diagnosing Subclinical Periodontitis in Adolescents.

This cross-sectional study compares the effectiveness of an active MMP-8 (aMMP-8) point-of-care (PoC)/chairside mouthrinse test to the conventional bleeding on probing (BOP) (cutoff 20%) test in detecting subclinical periodontitis/pre-periodontitis in Finnish adolescents. The study was carried out at the Kotka Health Center, Finland. A total of 47 adolescents (30 boys/17 girls) aged 15–17 were first tested with the aMMP-8 PoC test, followed by a full-mouth evaluation of clinical parameters of oral health including periodontal, oral mucosal, and caries assessment. A periodontist performed these clinical examinations. The aMMP-8 PoC test result had much stronger association with subclinical periodontitis than the BOP 20% test (2.8–5.3 times stronger in terms of odds ratio). The aMMP-8 PoC test had ≥2 times higher sensitivity than the BOP 20% test with, generally, the same specificity. Further, the aMMP-8 PoC test had generally better accuracy and lower false negative percentages. The aMMP-8 PoC test seemed to be more effective than the conventional BOP test in detecting subclinical periodontitis/pre-periodontitis in adolescents reducing the risk of their undertreatment. However, the sample size may be a limiting factor, and more studies are needed to confirm our results for both adolescents and adults.

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.

Validation of Matrix Metalloproteinase-9 (MMP-9) as a Novel Target for Treatment of Diabetic Foot Ulcers in Humans and Discovery of a Potent and Selective Small-Molecule MMP-9 Inhibitor That Accelerates Healing.

Diabetic foot ulcers (DFUs) are a significant health problem. A single existing FDA-approved drug for this ailment, becaplermin, is not standard-of-care. We previously demonstrated that upregulation of active matrix metalloproteinase (MMP)-9 is the reason that the diabetic wound in mice is recalcitrant to healing and that MMP-8 participates in wound repair. In the present study, we validate the target MMP-9 by identifying and quantifying active MMP-8 and MMP-9 in human diabetic wounds using an affinity resin that binds exclusively to the active forms of MMPs coupled with proteomics. Furthermore, we synthesize and evaluate enantiomerically pure ( R)- and ( S)-ND-336, as inhibitors of the detrimental MMP-9, and show that the ( R)-enantiomer has superior efficacy in wound healing over becaplermin. Our results reveal that the mechanisms of pathology and repair are similar in diabetic mice and diabetic humans and that ( R)-ND-336 holds promise for the treatment of DFUs as a first-in-class therapeutic.

Expression of active matrix metalloproteinase-9 as a likely contributor to the clinical failure of aclerastide in treatment of diabetic foot ulcers

Chronic wounds are a complication of diabetes. Treatment for diabetic foot ulcers is complex with little clinical recourse, resulting in 108,000 lower-limb amputations annually in the United States alone. Matrix metalloproteinases (MMPs) play important roles in the pathology and in the repair of chronic wounds. We previously identified active MMP-8 and MMP-9 in wounds of diabetic mice and determined that MMP-8 accelerates wound repair, while MMP-9 is the culprit for the diabetic wound being refractory to healing. Aclerastide, a peptide analog of angiotensin II, recently failed in phase III clinical trials for treatment of diabetic foot ulcers. We demonstrate herein that treatment of wounds of diabetic mice with aclerastide results in elevated levels of reactive oxygen species and of active MMP-9, which is likely an important contributor to the failure of aclerastide in clinical trials.

(R)-ND-336 as a Therapeutic Agent for Diabetic Wound Healing

Approximately 25% of diabetics develop diabetic foot ulcers (DFUs), resulting in 73,000 lower-limb amputations annually in the United States, with one-year prognosis of 50% mortality. While the involvement of matrix metalloproteinases (MMPs) in wound healing has been known, the challenge remained as to which MMP(s) among the 24 known human variants plays a role in the ailment, compounded by the fact that MMPs exist in three forms, the latent, the activated in complex with protein inhibitors TIMPs, and the activated uninhibited form. The first two are catalytically incompetent. With the use of an affinity resin that fishes out only the catalytically competent MMPs from the complex proteome, we identified active MMP-8 and MMP-9 in wounds of db/db mice and determined that MMP-9 is detrimental and that MMP-8 plays a beneficial role in wound healing. To demonstrate parallels in expression profiles in humans, we studied the debridement tissue from diabetic patients using the affinity resin coupled with proteomics. We identified the very same active MMP-8 and -9 in diabetic human wounds, indicating that the mechanisms of pathology and repair are similar in humans and in mice. Twenty-fold higher levels of the detrimental active MMP-9 were found in Wagner grade 1-4 wounds compared to control. These increased levels for MMP-9 are indeed significant, considering that active MMP-9 is a catalyst that contributes to the recalcitrance of wounds to heal. This observation makes MMP-9 an important target for inhibition with translational potential. We report herein that (R)-ND-336, a novel potent and selective inhibitor of MMP-9, accelerates healing of diabetic wounds and holds promise as recourse in treatment of DFUs. We note that NF-κB levels correlate with that of active MMP-9, hence it is a useful biomarker in identification of candidates for the treatment option. Disclosure M. Chang: None.

Oral fluid matrix metalloproteinase (MMP)-8 as a diagnostic tool in chronic periodontitis

Oral fluid matrix metalloproteinase (MMP)-8 as a diagnostic tool in chronic periodontitis Patricia Hernández-Ríos,1 Marcela Hernández,2,3 Mauricio Garrido,1 Taina Tervahartiala,4 Jussi M Leppilahti,4 Heidi Kuula,4 Anna Maria Heikkinen,4 Päivi Mäntylä,4 Nilminie Rathnayake,5 Solomon Nwhator,6 Timo Sorsa4,5 1Department of Conservative Dentistry, 2Department of Pathology and Oral Medicine, 3Laboratory of Periodontal Biology, Department of Conservative Dentistry, Faculty of Dentistry, University of Chile, Santiago, Chile; 4Department of Oral and Maxillofacial Diseases, Institute of Dentistry, Helsinki University and Helsinki University Central Hospital, Helsinki, Finland; 5Division of Periodontology, Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden; 6Department of Preventive and Community Dentistry, Faculty of Dentistry, College of Health Sciences, Obafemi Awolowo University, Ife, Nigeria Abstract: Periodontal diseases that affect the marginal and apical periodontium result from the interaction between bacterial biofilm and the host response. Oral fluid biomarkers might aid clinical diagnosis. Matrix metalloproteinases (MMPs) are a family of 24 proteases that act in physiological and pathological conditions. They can degrade almost all extracellular matrix constituents and regulate inflammatory processes. They are mainly inhibited by tissue inhibitors of metalloproteinases. The aim of this study was to perform a current literature review with a special reference on the diagnostic and clinical utility of oral fluid MMPs, especially MMP-8, and their inhibitors in periodontal and oral diseases. MMP-8 is the main collagenolytic MMP detected in oral fluids, such as saliva, oral mouthrinse, gingival crevicular fluid, and peri-implant fluid. MMP-8, and potentially MMP-9, in oral fluids represent strong biomarker candidates associated especially with periodontal disease diagnosis, severity, progression, and follow-up. Additionally, they show diagnostic potential for systemic conditions, such as pregnancy, myocardial infarction, and smoking. A commercially available mouthrinse, active MMP-8 chair-side/point-of-care lateral flow immunoassay, shows enough sensitivity and specificity to detect clinical signs of periodontitis. The current literature supports that high MMP-8 levels reflect the loss of periodontal supporting tissues rather than inflammation, representing a potentially useful side-diagnostic point-of-care oral disease biomarker, especially in periodontal diseases. Keywords: gingival crevicular fluid, saliva, biomarkers

Non-canonical role of matrix metalloprotease (MMP) in activation and migration of hepatic stellate cells (HSCs)

AimsMatrix metalloproteinases (MMPs) that degrade extracellular matrix (ECM) and help to resolve the excess matrix are considered to be under-expressed in fibrosis. MMPs are generally anti-fibrotic, however others can have pro-fibrotic functions. Therefore, the aim of this study was to find out the mechanism of pro-fibrotic function of MMPs in hepatic stellate cells' (HSC's) activation and migration. Main methodsHuman MMP Antibody Array from Abcam was used to profile MMPs in macrophages. Gelatin or casein zymography was performed using 10% SDS-polyacrylamide gels (SDS-PAGE) containing gelatin (1mg/ml) or Casein (1mg/ml) as substrate. HSCs migration assay was performed using Boyden chamber as described previously (Guo et al., 2007, McGarrigle et al., 2006, Shan et al., 2006 and Yang and Huang, 2005). Real-time PCR with SYBR green was performed using iTaq™ universal SYBR® Green supermix (BIO-RAD) and a 7500 Real-Time PCR System (Applied Biosystems). Collagen, type I, alpha 1 (COL1A1), alpha smooth muscle actin (α-SMA) expression was determined by immunoblot analysis. Key findingsWe first profiled the expression of all MMPs in primary murine bone marrow-derived macrophages (BMDMs) and differentiated THP-1 cells and found that MMP-8, -10, & -13, were significantly overexpressed after 12h of lipopolysaccharide (LPS) treatment. Based on this pattern of expression, we speculated that macrophage MMP-8,-10, &-13 might play a non-canonical role in HSCs activation. Further, we found that exogenous active MMP-8 (Collagenase-2) treated HSC shows markedly increased migration and COL1A1 expression as compared to MMP-10 and MMP-13 treated HSCs. Thus, macrophage MMP-8 (Collagenase-2) expression in macrophages emerges as an important moderator of HSC cell migration and invasion. SignificanceThese findings suggest that macrophage MMP-8 promotes HSC activation and might have a role in liver disease progression. MMP-8 targeting in the liver may have therapeutic potential in alcoholic liver disease (ALD).

Pilot Study on Oral Health Status as Assessed by an Active Matrix Metalloproteinase‐8 Chairside Mouthrinse Test in Adolescents

Matrix metalloproteinase (MMP)-8 is a major destructive collagenase involved in periodontitis and can be regarded as a periodontitis biomarker. A neutrophil collagenase 2 (active MMP-8 [aMMP-8]) oral fluid immunoassay has recently been demonstrated to be a periodontitis risk indicator among adults. The aim of this study is to investigate whether a point-of-care mouthrinse test based on an aMMP-8 immunoassay could identify patients with oral inflammatory burden (periodontitis and caries) among adolescents with early pathologic findings. This cross-sectional study was carried out at the Kotka Health Center, Finland. First, the aMMP-8 chairside mouthrinse test was performed on enrolled individuals (adolescents aged 15 to 17 years, n = 47), and the results were read based on a color change within 5 minutes. Then, full-mouth clinical parameters of oral health were assessed, including periodontal, oral mucosal, and caries status. The sensitivity and specificity of the test for bleeding on probing were 71.8% and 77.5%, respectively (P = 0.05); for ≥1 site with probing depth (PD) ≥4 mm, 48.3% and 100% (P <0.001); for ≥2 sites with PD ≥4 mm, 63.6% and 100% (P <0.001); and for >2 sites with PD ≥4 mm, 76.5% and 96.7% (P <0.01). Regarding periodontitis (≥1 site with PD ≥4 mm), hardly any false-positive results were identified. The sensitivity of the immunoassay for ≥1 caries lesions was 76.5%, and the specificity was 96.7% (P <0.01). In 5 minutes, the aMMP-8 chairside test showed promising results, recognizing oral inflammatory burden in adolescents with early initial signs of periodontitis. Caries lesions could also be detected, but less efficiently.