aMMP-8 POCT vs. Other Potential Biomarkers in Chair-Side Diagnostics and Treatment Monitoring of Severe Periodontitis.

This study aimed to compare several potential mouthrinse biomarkers for periodontitis including active matrix-metalloproteinase-8 (aMMP-8), total MMP-8, and other inflammatory biomarkers in diagnosing and monitoring the effects of nonsurgical periodontal therapy. Thirteen patients with stage III/IV periodontitis were recruited, along with thirteen periodontally and systemically healthy controls. These 13 patients were representative of the number of outpatients visiting any dentist in a single day. Full-mouth clinical periodontal parameters and biomarkers (the aMMP-8 point-of-care-test [POCT], total MMP-8, tissue inhibitor of MMPs (TIMP)-1, the aMMP-8 RFU activity assay, Myeloperoxidase, PMN elastase, calprotectin, and interleukin-6) were recorded at baseline and after nonsurgical therapy at 6 weeks. The aMMP-8 POCT was the most efficient and precise discriminator, with a cut-off of 20 ng/mL found to be optimal. Myeloperoxidase, MMP-8's oxidative activator, was also efficient. Following closely in precision was the aMMP-8 RFU activity assay and PMN elastase. In contrast, the total MMP-8 assay and the other biomarkers were less efficient and precise in distinguishing patients with periodontitis from healthy controls. aMMP-8, MPO, and PMN elastase may form a proteolytic and pro-oxidative tissue destruction cascade in periodontitis, potentially representing a therapeutic target. The aMMP-8 chair-side test with a cut-off of 20 ng/mL was the most efficient and precise discriminator between periodontal health and disease. The aMMP-8 POC test can be effectively used by dental professionals in their dental practices in online and real-time diagnoses as well as in monitoring periodontal disease and educating and encouraging good oral practices among patients.

Radiotherapy Increases aMMP-8-Levels and Neutrophil/Lymphocyte Ratio Rapidly in Head and Neck Cancer Patients: A Pilot Study.

Radiotherapy for head and neck carcinoma (HNC) has both curative and palliative purposes. This study investigated mouthrinse aMMP-8 levels, molecular forms of MMP-8, blood neutrophil counts and neurophil/lymphocyte ratios before and 3 weeks after HNC radiotherapy started. Thirteen HNC patients undergoing radiotherapy were included. Mouthrinse samples (before and 3 weeks after HNC radiotherapy had started) were assayed quantitatively by aMMP-8 point-of-care-kit (PerioSafe®/ORALyzer®) and by western immunoblot. Total neutrophil counts and neutrophil/lymphocyte ratios were evaluated in the hemogram results. Three weeks after HNC radiotherapy started, significant increases in aMMP-8 levels and neutrophil/lymphocyte ratios were observed. No significant difference was found in total neutrophil counts. Elevations of the activated and fragmented MMP-8 levels after HNC radiotherapy application were observed on western immunoblot analysis. The increase in the aMMP-8 levels and neutrophil/lymphocyte ratios indicate inflammation both locally and systemically suggesting increased risk for periodontitis due to the HNC radiotherapy.

Accuracy of aMMP-8 point-of-care test in indicating periodontal treatment outcomes in stage III/IV periodontitis: A 24-week follow-up study.

OBJECTIVE: To analyse the correspondence between aMMP-8 PoC test results and the clinical endpoints of non-surgical periodontal treatment in stage III/IV periodontitis. BACKGROUND: The diagnostic success of the active-matrix metalloproteinase-8 (aMMP-8) point-of-care (PoC) test has been demonstrated in various studies, but the evidence of its accuracy following periodontal treatment is limited. MATERIALS AND METHODS: Altogether 42 stage III/IV grade C periodontitis patients were included in this prospective diagnostic study. Clinical periodontal indices were recorded, aMMP-8 PoC test was applied and mouthrinse was collected before and at 6, 12 and 24 weeks after non-surgical periodontal treatment. Quantitative aMMP-8 levels were determined with immunofluorometric assay (IFMA) for the verification of the PoC test results. The accuracy of the aMMP-8 PoC test was assessed using previously established clinical endpoints as references. RESULTS: Sensitivity and specificity of aMMP-8 PoC test to indicate clinical endpoints were ranged as follows: Sensitivity 71.4% at baseline, 39.3%-42.4% at week 6, 28.6%-32.4% at week 12 and 35.3%-42.9% at week 24; specificity 64.3%-80% at week 6, 40%-57.1% at week 12 and 56%-64.3% at week 24. CONCLUSIONS: The accuracy of aMMP-8 PoC test in identifying clinical endpoints after non-surgical periodontal treatment is reduced in relation to baseline. Individual healing patterns of each diseased pocket eventually limit the accuracy of the dichotomous aMMP-8 oral rinse test during the post-treatment period.

Repeated Home-Applied Dual-Light Antibacterial Photodynamic Therapy Can Reduce Plaque Burden, Inflammation, and aMMP-8 in Peri-Implant Disease-A Pilot Study.

Until now, in clinical dentistry, antibacterial photodynamic therapy (aPDT) has been restricted to in-office treatments, which hampers repeated applications. This pilot study tested the benefit of a commercially available Lumoral® device designed for regular periodontal dual-light aPDT treatment at home. Seven patients with peri-implant disease applied dual-light aPDT daily in addition to their normal dental hygiene for four weeks. A single Lumoral® treatment includes an indocyanine green mouth rinse followed by 40 J/cm2 radiant exposure to a combination of 810 nm and 405 nm light. A point-of-care analysis of active-matrix metalloproteinase (aMMP-8), visible plaque index (VPI), bleeding on probing (BOP), and peri-implant pocket depth (PPD) measurements was performed on day 0, day 15, and day 30. Reductions in aMMP-8 (p = 0.047), VPI (p = 0.03), and BOP (p = 0.03) were observed, and PPD was measured as being 1 mm lower in the implant (p = ns). These results suggest a benefit of regular application of dual-light aPDT in peri-implantitis. Frequently repeated application can be a promising approach to diminishing the microbial burden and to lowering the tissue destructive proteolytic and inflammatory load around dental implants. Further studies in larger populations are warranted to show the long-term benefits.

Ability of matrix metalloproteinase-8 biosensor, IFMA, and ELISA immunoassays to differentiate between periodontal health, gingivitis, and periodontitis.

OBJECTIVE: The aim of this study was to determine the diagnostic utility of an MMP-8 biosensor assay in differentiating periodontal health from gingivitis and periodontitis and compare it with an established time-resolved immunofluorescence assay (IFMA) and enzyme-linked immunosorbent assay (ELISA). BACKGROUND: Currently available antibody-based assays display a wide variability in their ability to accurately measure matrix metalloproteinase-8 (MMP-8) levels in saliva. METHODS: Salivary MMP-8 levels were analyzed in 189 systemically healthy participants using an antibody-based biosensor prototype that operates using a surface acoustic wave technology and compared with IFMA and ELISA antibody assays. Participants were categorized into 3 groups: periodontal health (59), gingivitis (63), and periodontitis (67). A sub-population of participants (n = 20) with periodontitis received periodontal treatment and were monitored for 6 months. RESULTS: All the assays demonstrated significantly higher salivary MMP-8 concentrations in participants with periodontitis versus gingivitis, periodontitis versus health, and gingivitis versus health (all p < .05). The biosensor data demonstrated significant correlations with IFMA (r = .354, p < .001) and ELISA (r = .681, p < .001). Significant reductions in salivary MMP-8 concentrations were detected by the biosensor (p = .030) and IFMA (p = .002) in participants with periodontitis 6 months after non-surgical periodontal treatment. IFMA had the best sensitivity (89.2%) for detecting periodontitis and gingivitis versus health and 96.6% for detecting periodontitis versus health and gingivitis. The biosensor had an AUC value of 0.81 and diagnostic accuracy of 74.2% for differentiating periodontitis and gingivitis from health; an AUC value of 0.86 and diagnostic accuracy of 82.8% for periodontitis versus health and gingivitis. CONCLUSIONS: The biosensor, IFMA, and ELISA assays differentiated between periodontal health, gingivitis, and periodontitis based on salivary MMP-8 levels. Only the biosensor and, particularly, IFMA identified an effect of periodontal treatment in the participants with periodontitis. Our findings support the potential utility of salivary oral fluid aMMP-8-based point-of-care technology in the future of periodontal diagnostics.

Active matrix metalloproteinase-8 (aMMP-8) point-of-care test (POCT) in the COVID-19 pandemic.

INTRODUCTION: Active matrix metalloproteinase (aMMP)-8 utilized in point-of-care testing (POCT) is regarded as a potential biomarker for periodontal and peri-implant diseases. Various host and microbial factors eventually influence the expression, degranulation, levels and activation of aMMP-8. The type of oral fluids (saliva, mouthrinse, gingival crevicular, and peri-implant sulcular fluids [GCF/PISF], respectively) affect the analysis. AREAS COVERED: With this background, we aimed to review here the recent studies on practical, inexpensive, noninvasive and quantitative mouthrinse and GCF/PISF chair-side POCT lateral flow aMMP-8 immunoassays (PerioSafe and ImplantSafe/ORALyzer) and how they help to detect, predict, monitor the course, treatment and prevention of periodontitis and peri-implantitis. The correlations of aMMP-8 POCT to other independent and catalytic activity assays of MMP-8 are also addressed. EXPERT OPINION: The mouthrinse aMMP-8 POCT can also detect prediabetes/diabetes and tissue destructive oral side-effects due to the head and neck cancers' radiotherapy. Chlorhexidine and doxycycline can inhibit collagenolytic human neutrophil and GCF aMMP-8. Furthermore, by a set of case-series we demonstrate the potential of mouthrinse aMMP-8 POCT to real-time/online detect periodontitis as a potential risk disease for coronavirus disease 2019 (COVID-19). The clinical interdisciplinary utilization of aMMP-8 POCT requires additional oral, medical, and interdisciplinary studies.

Low association between bleeding on probing propensity and the salivary aMMP-8 levels in adolescents with gingivitis and stage I periodontitis.

BACKGROUND AND OBJECTIVE: Bleeding on probing (BOP) is a widely accepted measure used in periodontal diagnostics. Previous studies suggest that several factors can affect BOP propensity. The aim of this study was to investigate the relative impact of different local and modifying factors on BOP levels. MATERIALS AND METHODS: The oral health of five hundred and forty-four adolescents (two birth cohorts) aged 15-17 years living in Kotka, Finland, was examined including periodontal probing depth, visible plaque index, root calculus, and BOP. Whole saliva samples were collected and measured for active matrix metalloproteinase-8 (aMMP-8) by time-resolved immunofluorometric assay (IFMA). RESULTS: Bacterial plaque/calculus accumulation (oral hygiene) had a major influence on BOP levels. The relative impact was several times greater compared with the extent of periodontal pocketing, aMMP-8 levels, smoking, toothbrushing, or gender. Furthermore, BOP levels were significantly elevated among adolescents with poor oral hygiene than good oral hygiene even if adjusted for the extent of periodontal pocketing (P < .001). BOP levels could be low even if several ≥ 4 mm deep periodontal pockets existed. The difference in the extent of periodontal pocketing was not significant between the two birth cohorts of adolescents (P = .731). CONCLUSIONS: BOP levels can be regarded as an important indicator of the extent of bacterial challenge and its adverse effects on the gingival inflammation. However, the level of oral hygiene may mask the association between the extent of gingival bleeding and the severity of the periodontal inflammatory condition. Thus, relying on BOP levels (below 10% or 20%) may provide insufficient information about the periodontal treatment need of an adolescent depending on his/her level of oral hygiene. Yet, more research is needed to confirm the results, also in adult populations.

Active matrix metalloproteinase-8 and interleukin-6 detect periodontal degeneration caused by radiotherapy of head and neck cancer: a pilot study.

Background: This cohort study investigated the role of the active matrix metalloproteinase-8 (aMMP-8) and interleukin-6 (IL-6) as oral fluid biomarkers for monitoring the periodontal degeneration occurring in head and neck cancer (HNC) patients treated by radiotherapy. Research design and methods: Eleven patients, aged 28-74, diagnosed with HNC were included in the study. Complete periodontal and oral examinations were performed pre-radiotherapy and 1 month after radiotherapy. Mouthrinse samples (pre-radiotherapy, after 6 weeks of radiotherapy and 1 month after radiotherapy) were assayed by aMMP-8 point-of-care-kit (PerioSafe®/ORALyzer®) for aMMP-8 and ELISA for IL-6. Results: HNC radiotherapy had a deteriorating impact on the periodontium and a significant impact on periodontal biomarkers aMMP-8 and IL-6 and increased their levels in mouthrinse. Clinical-attachment-loss (CAL) (site of greatest loss: mean = 1.7 mm, range = 1-3 mm) corresponding to rapid progression of periodontitis. There was a positive repeated measures correlation (rmcorr = 0.667) between the aMMP-8 and IL-6 levels. Conclusions: Elevated aMMP-8 levels were observed 1 month after radiotherapy among some HNC patients suggesting a prolonged increased susceptibility to further periodontal tissue destruction. Currently available aMMP-8 point-of-care testing could be useful to monitor and assess quantitatively online and real-time the risk of deterioration of periodontal health during HNC radiotherapy.

Oral Anti-Inflammatory and Symbiotic Effects of Fermented Lingonberry Juice-Potential Benefits in IBD.

Microbial dysbiosis may manifest as inflammation both orally and in the gastrointestinal tract. Altered oral and gut microbiota composition and decreased diversity have been shown in inflammatory bowel disease (IBD) and periodontal disease (PD). Recent studies have verified transmission of oral opportunistic microbes to the gut. Prebiotics, probiotics, or dietary interventions are suggested to alleviate IBD symptoms in addition to medicinal treatment. Lingonberries contain multiple bioactive molecules, phenolics, which have a broad spectrum of effects, including antimicrobial, anti-inflammatory, antioxidant, anti-proteolytic, and anti-cancer properties. An all-natural product, fermented lingonberry juice (FLJ), is discussed as a potential natural anti-inflammatory substance. FLJ has been shown in clinical human trials to promote the growth of oral lactobacilli, and inhibit growth of the opportunistic oral pathogens Candida, Streptococcus mutans, and periodontopathogens, and decrease inflammation, oral destructive proteolysis (aMMP-8), and dental microbial plaque load. Lactobacilli are probiotic and considered also beneficial for gut health. Considering the positive outcome of these oral studies and the fact that FLJ may be swallowed safely, it might be beneficial also for the gut mucosa by balancing the microbiota and reducing proteolytic inflammation.

Diagnostic value of aMMP-8 and azurocidin in peri-implant sulcular fluid as biomarkers of peri-implant health or disease.

OBJECTIVE: The objective of this study was to investigate the effectiveness of testing for active matrix metalloproteinase-8 (aMMP-8) by a quantitative point-of-care (PoC), chairside lateral flow immunotest and azurocidin, in the peri-implant sulcular fluid (PISF), as biomarkers for the presence or absence of peri-implant diseases. BACKGROUND: Current research indicates that proinflammatory cytokines and extracellular matrix-degrading enzymes may be of value to diagnose and predict peri-implant disease initiation and progression, but more data are needed. METHODS: Eighty patients with implants were recruited. PISF samples were collected and quantitatively analyzed for aMMP-8 (chairside) and azurocidin with ELISA. Radiographic assessments and clinical indices (probing depth, probing attachment level, bleeding on probing, and plaque) were recorded after sampling. Kruskal-Wallis test and pairwise post hoc Dunn-Bonferroni test were used to relate aMMP-8 levels and azurocidin levels to clinical parameters. The diagnostic ability of aMMP-8 (ng/mL) and azurocidin was analyzed by receiver operator curve analysis. Area under the curve (AUC) was calculated and the Spearman's rho, and the coefficient of determination (R2) were used to calculate the correlations between aMMP-8, azurocidin, and periodontal parameters. RESULTS: Statistically significant differences were observed for aMMP-8 levels but not for azurocidin between healthy implants, implants with mucositis, and those with peri-implantitis (13.65 ± 7.18, 32.33 ± 21.20, and 73.07 ± 43.93 ng/mL, respectively), (Kruskall-Wallis test p < .05). The aMMP-8 test with a threshold of 20 ng/mL has a sensitivity of 71.7% and a specificity of 77.8% to identify peri-implantitis and healthy implants, respectively. AUC was found to be 0.814, and the accuracy of the method reaches 73.8%. Above a cutoff value of 33.7 ng/mL of aMMP-8, the accuracy of the test to detect peri-implantitis reaches 77.5% in relation to 62.5% of BoP from the same site. CONCLUSION: Taken collectively, present data indicate that the aMMP-8 PoC lateral flow immunotest can be a beneficial, adjunctive diagnostic quantitative tool for real-time screening for peri-implant diseases.

Lingonberry (Vaccinium vitis-idaea L.) Fruit Phenolic Bioactivities-A Review of In Vitro and In Vivo Human Studies.

This review is focused on the effects of lingonberry (Vaccinium vitis-idaea L.) fruit phenolic compounds in human in vitro cells and in vivo clinical studies. Studies with lingonberries, lingonberry juice/lingonberry nectar/fermented lingonberry juice, and phenolic fractions with active molecules are reviewed. Lingonberry's bioactive substances have a diverse range of antimicrobial, anti-inflammatory, antiproteolytic, anticancer, and antioxidant properties. Fermentation of lingonberries and modulation of the dysbiotic microbiome to a more symbiotic composition by favoring the growth of lactobacilli and inhibiting the growth of human opportunistic pathogens are discussed. Research results suggest that more studies on humans are needed.

Prediabetes Associates with Matrix Metalloproteinase-8 Activation and Contributes to the Rapid Destruction of Periodontal Tissues.

OBJECTIVE: The aim of this cross-sectional study was to investigate the relationship between periodontitis, potential periodontitis oral fluid biomarkers, and prediabetes. MATERIALS AND METHODS: This study included 150 Greek adults aged 25 to 78 years who were tested with an Hemoglobin A1C (HBA1c) diagnostic system, an active-matrix metalloproteinase-8 (aMMP-8) point-of-care (PoC) test, and several salivary biomarkers enzyme-linked immunosorbent assay tests and gelatin zymography. A full-mouth clinical examination was performed to assess their periodontal and oral health status. STATISTICAL ANALYSIS: The Kruskal-Wallis test was used to determine the statistically significant difference in the levels of periodontal oral fluid biomarkers between the different periodontitis stages, periodontitis grades, and the stages and grades of periodontitis combined. Spearman's rank correlation was performed to assess the strength and direction of the association between aMMP-8 and HbA1c levels (<5.7 and ≥5.7%) and with the other oral fluid biomarkers among patients with severe periodontitis. A two-sided p-value below 0.05 was considered statistically significant in this study. RESULTS: aMMP-8, but not total MMP-8 or other biomarkers, associated significantly with the stage and grade of periodontitis combined (p < 0.001, Kruskal-Wallis test). Among stage III grade C periodontitis patients, aMMP-8 levels were significantly positively correlated with prediabetes (Spearman's rho = 0.646, p = 0.044), total MMP-8 (rho = 0.636, p = 0.048), PMN Elastase (rho = 0.729, p = 0.017), total MMP-9 (rho = 0.721, p = 0.019), and total MMP-8/TIMP-1 molar ratio (rho = 0.879, p < 0.001). CONCLUSION: Prediabetic disease development can upregulate MMP-8 expression (total MMP-8) in rapidly progressing, severe periodontitis, where MMP-8 latent species are further activated into their active forms (aMMP-8). Simultaneously, several proinflammatory biomarker levels are elevated in this tissue-destructive biomarker cascade. This development is easily detectable online/in real-time within 5 minutes by aMMP-8 PoC testing at the dentist's office.

In Vivo Regulation of Active Matrix Metalloproteinase-8 (aMMP-8) in Periodontitis: From Transcriptomics to Real-Time Online Diagnostics and Treatment Monitoring.

BACKGROUND: This study investigated in vivo regulation and levels of active matrix metalloproteinase-8 (aMMP-8), a major collagenolytic protease, in periodontitis. METHODS: Twenty-seven adults with chronic periodontitis (CP) and 30 periodontally healthy controls (HC) were enrolled in immunohistochemistry and transcriptomics analytics in order to assess Treponema denticola (Td) dentilisin and MMP-8 immunoexpression, mRNA expression of MMP-8 and its regulators (IL-1ß, MMP-2, MMP-7, TIMP-1). Furthermore, the periodontal anti-infective treatment effect was monitored by four different MMP-8 assays (aMMP-8-IFMA, aMMP-8-Oralyzer, MMP-8-activity [RFU/minute], and total MMP-8 by ELISA) among 12 CP (compared to 25 HC). RESULTS: Immunohistochemistry revealed significantly more Td-dentilisin and MMP-8 immunoreactivities in CP vs. HC. Transcriptomics revealed significantly elevated IL-1ß and MMP-7 RNA expressions, and MMP-2 RNA was slightly reduced. No significant differences were recorded in the relatively low or barely detectable levels of MMP-8 mRNAs. Periodontal treatment significantly decreased all MMP-8 assay levels accompanied by the assessed clinical indices (periodontal probing depths, bleeding-on-probing, and visual plaque levels). However, active but not total MMP-8 levels persisted higher in CP than in periodontally healthy controls. CONCLUSION: In periodontal health, there are low aMMP-8 levels. The presence of Td-dentilisin in CP gingivae is associated with elevated aMMP-8 levels, potentially contributing to a higher risk of active periodontal tissue collagenolysis and progression of periodontitis. This can be detected by aMMP-8-specific assays and online/real-time aMMP-8 chair-side testing.

Antimicrobial and Anti-Inflammatory Oral Effects of Fermented Lingonberry Juice-A One-Year Prospective Human Intervention Study.

OBJECTIVES: A 1-year prospective human intervention study was performed to examine the anticaries, anti-inflammatory, antiproteolytic, and antimicrobial effects of fermented lingonberry juice (FLJ), used as a mouthwash for a period of 6 months, followed by a 6-month washout period. MATERIALS AND METHODS: Twenty-five adults were recruited from private dental clinics in Helsinki and Joensuu (Finland). Standard oral examinations and sample gatherings were performed at base level, 6 months, and 1 year for oral Streptococcus mutans (S. mutans), Candida, and Lactobacilli levels, and active matrix metalloprotease-8 (aMMP-8) levels, and for decayed, missing, filled teeth (DMFT), decayed, missing filled surfaces (DMFS) and decayed surfaces (DS) indexes, and probing pocket depths (PPDs), bleeding on probing (BOP), and visible plaque index (VPI). FLJ was used by the participants once daily for 30 seconds for 6 months. FLJ contains 0.212% (w/v) polyphenols, 3% (w/ v) sugars, and contains no excipients. Ten milliliters of FLJ were equal to 1 dL of lingonberry juice. STATISTICAL ANALYSIS: Statistical analyses were performed with nonparametric Friedman's test and pairwise post-hoc analysis with Dunn-Bonferroni test, SPSS (version 27; IBM) and p < 0.05 was considered as statistically significant. RESULTS: The levels of S. mutans and Candida counts, DS, BOP, and VPI decreased significantly (p < 0.05) during the FLJ period. Lactobacilli counts increased significantly, while there was also significant difference in aMMP-8 levels, DMFT, and DMFS between the three measurement points. PPDs were not affected. CONCLUSIONS: The specially formulated FLJ may have a positive decreasing effect on S. mutans, and Candida counts as well as decrease low-grade inflammation and proteolytic burden in the oral mucosa and periodontal tissues. The beneficial effects to the oral cavity of FLJ mouthwash may be useful among patients with oral diseases, such as dental caries, periodontitis and candidosis.