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Objective To study the effect of orally administered Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus rhamnosus GG on the salivary levels of Matrix Metalloproteinases (MMP)-8, MMP-9 and of Tissue Inhibitor of Metalloproteinases (TIMP)-1 in healthy adults. Furthermore, the correlations between MMP-8, MMP-9 and TIMP-1 and plaque and gingival indices, salivary mutans streptococci and lactobacilli counts, and stimulated saliva secretion rate were analysed. Design The salivary samples originated from a randomized controlled trial where healthy student volunteers consumed probiotic or placebo lozenges twice a day for four weeks. The saliva samples were collected and clinical parameters measured at the baseline and at the end of the original study. For this study, the salivary levels of MMP-8, MMP-9 and TIMP-1 were analysed with immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay (ELISA). Results In the probiotic group (n = 29), salivary MMP-9 levels increased (p < 0.01) and TIMP-1 levels decreased (p < 0.01) significantly during the intervention. Furthermore, MMP-9/TIMP-1 ratio differed significantly from the baseline level (p < 0.01). These changes were not observed in the control group (n = 31). In the whole data, salivary MMP-9 and gingival index correlated (r = 0.260, p < 0.05 at baseline and r = 0.354, p < 0.01 at the end of the study). Intergroup differences or correlations with other clinical parameters were not found. Probiotic consumption did not affect the saliva flow rate. Conclusions Increased MMP-9 and decreased TIMP-1 levels in saliva may indicate that probiotics have immunomodulatory effects in the oral cavity. Furthermore, increased salivary MMP-9 levels may be an indication of the defensive potential of matrix metalloproteinases. [ABSTRACT FROM AUTHOR]

Objectives The present study aimed to investigate the effect of HNP-1 on the matrix metalloproteinase (MMP)-2, -8 and -9 secretions of two oral squamous cell carcinoma (OSCC) cell lines (UT-SCC-43A and UT-SCC-43B). Design In all experiments, the two OSCC cell lines were incubated with graded concentrations (0, 1, 5, and 10 μg/ml) of HNP-1 for 24 and 48 h. Cell viability was measured using a colorimetric proliferation test and cell death was analyzed with a colorimetric cytotoxicity detection kit. Enzyme activity of MMP-2 and MMP-9 was detected by using gelatin zymography, and molecular weight forms of MMP-8 were determined by Western-blot and a densitometric quantitation method. Results Both cell lines showed a significant increase in LDH toxicity at 24 h (UT-SCC-43A: p = 0.005 & UT-SCC-43B: p = 0.014). Reduced gelatinolytic activities of proMMP-2 were detected in UT-SCC-43B cell line after 24 and 48 h of incubation with HNP-1 (1 μg/ml: p < 0.001, 5 μg/ml: p < 0.001, and 10 μg/ml: p = 0.0225). MMP-8 levels of both cell lines decreased at 200–250 kDa after 24 h of incubation, while after 48 h only UT-SCC-43B decreased at 45–50 kDa. Conclusions Our results indicate that HNP-1 suppresses the secretion of MMP-2, -8, and -9 in OSCC cell lines. [ABSTRACT FROM AUTHOR]

Abstract: Matrix metalloproteinases (MMPs) are present in sound coronal dentin and may play a role in collagen network degradation in bonded restorations. We investigated whether these enzymes can also be detected in root dentin. Crown and root sections of human teeth were powderized, and dentin proteins were extracted by using guanidine-HCl and EDTA. Extracts were analyzed by zymography and Western blotting for matrix metalloproteinases detection. Zymography revealed gelatinolytic activities in both crown and root dentin samples, corresponding to MMP-2 and MMP-9. MMP-2 was more evident in demineralized root dentin matrix, whereas MMP-9 was mostly extracted from the mineralized compartment of dentin and presented overall lower levels. Western blot analysis detected MMP-8 equally distributed in crown and root dentin. Because MMPs are also present in radicular dentin, their contribution to the degradation of resin-dentin bonds should be addressed in the development of restorative strategies for the root substrate. [Copyright &y& Elsevier]

Abstract: Objective: Previously an unidentified collagenolytic metalloprotease together with gelatinase (matrix metalloproteinase-2, MMP-2), and enamelysin (MMP-20) have been detected in human dentin. The aim of the study was to characterize dentinal collagenolytic enzymes. Furthermore, we hypothesized that the dentinal MMPs are protected by the mineral phase, and studied the stability of dentinal MMPs. Design: To characterize dentinal collagenolytic enzymes, we used Western blotting with specific antibodies against MMP collagenases (MMP-1, -8, and -13) and cathepsin K. MMP-8 immunofluorometric assay (IFMA) was also used for MMP-8 detection, and functional collagenase activity was examined with type I collagen degradation assay. The stability of dentinal MMPs was examined by autoclaving dentin blocks before protein extraction and subsequent examination of protein levels and the activities of dentin collagenase and gelatinases. Results: MMP-8 (collagenase-2) was detected in dentin both with Western blot and IFMA, and dentinal samples also cleaved the intact type I collagen into characteristic 3/4(αA)-cleavage products in vitro. No other collagenases or cathepsin K were detected. In autoclaved samples no MMP-8 was found, but gelatinase activity was observed in protein fractions of mineralized dentin. Conclusions: MMP-8 represents the major collagenase in human dentin. Unlike MMP-8, dentinal gelatinases can be detected after autoclave treatment of dentin, indicating their high resistance to external sample treatment procedures. [Copyright &y& Elsevier]

Periodontal disease is a chronic multifactorial infectious and inflammatory disease associated with several chronic systemic diseases, such as diabetes, cardiovascular diseases (CVD), chronic obstructive pulmonary disease, hypertension, Alzheimer's disease and so on. These same systemic diseases have been associated with severe COVID-19 infections. Several recent studies have suggested hypotheses for the potential association between periodontal disease and severe COVID-19. Periodontal disease is also one of the most prevalent diseases globally. All this supports the importance of good oral health, also in the COVID-19 era. Thus, new strategies and approaches to identify patients at risk of periodontal disease could be beneficial to enhance secondary prevention, especially if targeted to COVID-19 risk groups. Diagnostic biomarkers for periodontal disease have been researched extensively. Potential biomarkers in oral fluid with currently available rapid non-invasive point-of-care technology, such as aMMP-8, could help to extend screening and identification of patients at risk for periodontal disease also to situations and places where professional dental expertise and equipment are limited or unavailable. i.e., nursing and care homes, and rural and distant places. The oral fluid point-of-care technologies could also be useful in the hands of medical professionals (diabetes, CVD, etc.) to identify patients at risk for undiagnosed periodontal disease and to refer them to a dentist for examination and evaluation. Finally, if there is a causality between periodontal disease and severe COVID-19 infections, these point-of-care oral fluid biomarker technologies could possibly also help in the assessment of the risk of deterioration and complications. [ABSTRACT FROM AUTHOR]

Abstract: Aim: The rationale of this study was to address whether local or systemic changes reflect proteolytic (matrix metalloproteinase-13) or oxidative (myeloperoxidase) stress in renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), in patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy. Material and methods: Gingival crevicular fluid (GCF) samples were collected from sites with (GO+) and without GO (GO−) in CsA patients having GO; GO− sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum myeloperoxidase (MPO) and matrix metalloproteinase-13 (MMP-13) levels were determined by ELISA. Results: GO+ sites in CsA patients having GO had elevated GCF MPO levels than those of CsA patients having no GO, tacrolimus and healthy subjects (p <0.005), but comparable to those of gingivitis. GCF MPO levels were higher in GO+ compared to GO− sites in CsA patients having GO (p <0.05). Patient groups had similar, but higher GCF MMP-13 levels than healthy group. Conclusions: These results show that CsA and tacrolimus therapy have not a significant effect on GCF MPO and MMP-13 levels, and gingival inflammation seems to be the main reason for their elevations. [ABSTRACT FROM AUTHOR]

Objectives This cross-sectional study aimed to investigate the relationship between thalassemia major (TM) and gingival inflammation through the salivary, serum, and gingival crevicular fluid (GCF) levels of matrix metalloproteinase (MMP)-8, MMP-9 and tissue inhibitor of MMP (TIMP)-1. Methods Biofluid samples and full-mouth clinical periodontal recordings were obtained from 29 otherwise healthy patients with TM and 25 systemically healthy (SH) individuals. Biofluid samples were evaluated by immunofluorometric assay (IFMA) and enzyme-linked immunoassays (ELISAs). Data were tested statistically by Kolmogorov Simirnov, Mann–Whitney U tests, Spearman correlation analysis. Results Age, smoking status, bleeding on probing, plaque index were similar in the study groups, but probing depth, gender data exhibited significant differences ( p = 0.037 for both). Salivary MMP-8, MMP-9, TIMP-1 concentrations were significantly higher in the TM than SH group ( p = 0.014; p < 0.001; p = 0.042, respectively). Serum TIMP-1 concentrations were significantly higher; MMP-8/TIMP-1, MMP-9/TIMP-1 molar ratios were significantly lower in the TM than SH group ( p < 0.001; p = 0.005; p = 0.022, respectively). Very few GCF samples revealed biochemical data above the detection limits. Numerous correlations were found between clinical periodontal parameters and biochemical data. Conclusions It may be suggested that TM may exacerbate the local inflammatory response as manifested in salivary MMP-8, MMP-9, TIMP-1 levels. [ABSTRACT FROM AUTHOR]

Objective Matrix metalloproteinases MMP-2 and MMP-9 have been associated with juvenile parotitis. However, the role of MMP-8 has not been addressed previously. This work focuses on salivary MMP-8 and -9 levels in juvenile parotitis. Methods During a five-year period at Helsinki University Hospital, a tertiary care hospital, 41 patients aged 17 or under, were identified as having parotitis; from 36 of these patients, saliva samples were collected for MMP-8 IFMA (time-resolved immunofluorometric assay) analyses. Control saliva samples were collected from 34 age- and gender-matched children admitted for an elective surgery who had no history of parotitis. For comparison, salivary levels of MMP-9, tissue inhibitor of matrix metalloproteinase (TIMP-1), MMP-8/TIMP-1 ratio, human neutrophil elastase (HNE), and myeloperoxidase (MPO) were analyzed by ELISA. Additionally, salivary MMP-8 levels were compared to historical saliva samples from 18 adult gingivitis patients as well as to 10 healthy adult controls. Results The median (25%, 75% percentile) MMP-8 concentration in saliva of parotitis patients was significantly lower than MMP-8 concentration in saliva of their controls [50.4 ng/ml (37.5, 72.9) vs. 148.5 ng/ml (101.2, 178.5) p < 0.0001] and lower than in patients with gingivitis [347.9 ng/ml (242.6, 383.2) p < 0.0001] or healthy adult controls [257.2 ng/ml (164.9, 320.7) p < 0.0001]. The MMP-8/TIMP-1 ratio was lower than in controls [0.13 (0.05–0.02) vs. 0.3 (0.17–0.46) p < 0.0001]. The median MMP-9 concentration in saliva of parotitis patients was significantly higher than in controls [143.9 ng/m (68.8–189.0) vs. 34.9 ng/ml (16.3–87.6) p < 0.0001]. Neither HNE, MPO, nor TIMP-1 alone separated the patients from the control groups. Conclusions MMP-9 was up-regulated in juvenile parotitis saliva, suggesting that MMP-9 may play a destructive role in juvenile parotitis, as others have suggested. The present novel findings reveal a decreased salivary MMP-8 concentration, suggesting that MMP-8 may reflect in juvenile parotitis down-regulated or anti-inflammatory immune characteristics. [ABSTRACT FROM AUTHOR]

Objectives Despite increasing evidence for an association of obstructive sleep apnea syndrome (OSAS) and periodontal disease, the pathophysiological linking mechanisms remain unclear. This study aims to evaluate the salivary and serum matrix metalloproteinase-2, -8, -9 (MMP-2, -8, -9), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), myeloperoxidase (MPO), neutrophil elastase (NE), neutrophil gelatinase-associated lipocalin (NGAL), as well as degree of activation of MMP-2, -9 of patients with and without OSAS. Design A total of 50 individuals were included in the study. There were 13, 17 and 20 individuals, respectively in the control (non-OSAS) group, mild-to-moderate OSAS and severe OSAS groups. Saliva, serum samples and clinical periodontal parameters were collected. Biofluid samples were analysed by immunofluorometric assay (IFMA), enzyme-linked immunosorbent assay (ELISA), western immunoblotting and gelatine zymography. Statistical analyses were performed using D’Agostino–Pearson omnibus normality test, Kruskal–Wallis test and Spearman rho rank correlation analysis. Results There were no statistically significant differences in clinical periodontal parameters between the study groups. Salivary NE and proMMP-2 levels were significantly lower in the OSAS groups than the control group ( p < 0.05). Serum proMMP-9 concentration and the degree of MMP-9 activation in saliva were significantly lower in the severe OSAS group than the control group ( p < 0.05). There were significant correlations between salivary and serum proMMP-9 and -2 concentrations ( p < 0.05). Serum proMMP-2, NE and salivary proMMP-9 and -2 negatively correlated with indicators of OSAS severity ( p < 0.05). Conclusions The present findings do not support a pathophysiological link between the severity of OSAS and clinical periodontal status via neutrophil enzymes or MMPs. [ABSTRACT FROM AUTHOR]

Summary: Cancer invasion induces extracellular matrix remodeling and collagen degradation. The aim of this study was to assess whether serum levels of type I and III collagen degradation products were associated with patient survival in head and neck squamous cell carcinoma (HNSCC). A novel enzyme immunoassay was developed for measuring type III collagen N-terminal telopeptide (IIINTP) in human serum samples. In addition, type I collagen C-terminal telopeptide (ICTP), matrix metalloprotease-8 (MMP-8) and tissue inhibitor of metalloproteases-1 (TIMP-1) were assessed in 205 blood samples from HNSCC patients. High levels of serum ICTP and IIINTP and plasma TIMP-1 were associated with poor survival. The concentration of ICTP was associated with levels of IIINTP and TIMP-1. The plasma concentration of MMP-8 was associated with tumor stage, but not with survival or levels of ICTP, IIINTP or TIMP-1 suggesting that other collagenases/proteases are responsible for the cleavage of type I and type III collagens. The rate of type I and type III collagen degradation is associated with patient survival and can be used as a prognostic marker in HNSCC. [ABSTRACT FROM AUTHOR]

Abstract: Aim: To study the systemic levels of matrix metalloproteinases (MMP) -7, -8 and -9 and their inhibitor TIMP-1 in cardiac arrest patients and the association with mild therapeutic hypothermia treatment on the serum concentration of these enzymes. Methods: MMP-7, -8 and -9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) were analysed in blood samples obtained from 51 patients resuscitated from cardiac arrest. The samples were taken at 24 and 48h from restoration of spontaneous circulation (ROSC). The biomarker levels were compared between patients (N =51) and healthy controls (N =10) and between patients who did (N =30) and patients who did not (N =21) receive mild therapeutic hypothermia. Results: MMP-7 (median 0.47ng/ml), MMP-8 (median 31.16ng/ml) and MMP-9 (median 253.00ng/ml) levels were elevated and TIMP-1 levels suppressed (median 78.50ng/ml) in cardiac arrest patients as compared with healthy controls at 24h from ROSC. Hypothermia treatment associated with attenuated elevation of MMP-9 (p =0.001) but not MMP-8 (p =0.02) or MMP-7 (p =0.69). Concentrations of MMPs -7, -8 and -9 correlated with the leukocyte count but not with C-reactive protein (CRP) or neurone-specific enolase (NSE) levels. Conclusion: We demonstrated that the systemic levels of MMP-7, -8 and -9 but not TIMP-1 are elevated in cardiac arrest patients in the 48h post-resuscitation period relative to the healthy controls. Patients who received therapeutic hypothermia had lower MMP-9 levels compared to non-hypothermia treated patients, which generates hypothesis about attenuation of inflammatory response by hypothermia treatment. [Copyright &y& Elsevier]

Inflammation causes epithelial cell sloughing and basement membrane (BM) exposure in canine pulmonary eosinophilia (PE). leading to degradation of the epithelial cell attachment component, laminin-5 γ2-chain, into small molecular weight fragments. The subsidence of inflammation after treatment down-regulates degradation. Laminin-5 γ2-chain levels and molecular forms in bronchoalveolar lavage fluid (BALF) were analysed semiquantitatively by Western immunoblotting to compare PE affected (n = 20) and healthy dogs (n = 16) as well as PE dogs (n = 6) before and after corticosteroid treatment. PE dogs expressed significantly elevated levels of total (P < 0.01), 36 kDa (P < 0.051 and 53 kDa (P < 0.05) laminin-5 γ2-fragments. The 36 kDa fragment decreased significantly (P < 0.05) after treatment. The laminin-5 γ2-chain degradation products may he linked to epithelial cell sloughing and BM exposure or healing. [ABSTRACT FROM AUTHOR]

Chymotrypsin-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. proMMP-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. Td-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. Regarding 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. [ABSTRACT FROM AUTHOR]

• Pregnant/postpartum women had higher saliva/GCF MMP-8 and MMP-9 than controls. • Gingivitis patients within groups had higher saliva/GCF MMP-8 and MMP-9 than controls. • Non-pregnant women exhibited lower TIMP-1 in saliva than the postpartum group. During pregnancy, mothers undergoe considerable physiological changes affecting the whole body including periodontal tissues. Susceptibility to gingival inflammation during pregnancy could be mediated by modulation of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). Therefore, the aim of this study was to investigate salivary and gingival crevicular fluid (GCF) levels of MMPs and TIMPs during the second and third trimester of pregnancy and postpartum. Saliva and GCF samples were collected from 96 pregnant women (PW) before and after giving birth. The sixty matched non-pregnant women (N-PW) were recruited as a control group and full-mouth periodontal examination was performed. The levels of MMP-8, MMP-9 and TIMP-1 were determined by immunofluorometric and enzyme-linked immunosorbent assays. The PW group exhibited significantly higher levels of MMP-8 and MMP-9 in their saliva than the N-PW group while corresponding salivary TIMP-1 levels were significantly lower in NPW compared to the postpartum stage. This resulted in significantly higher MMP-8/TIMP-1 and MMP-9/TIMP-1ratio in the saliva from PW before and after birth than in that from N-PW. MMP-8, MMP-9 and TIMP-1 levels were higher in GCF from PW and postpartum than in that from N-PW. MMP-8 and MMP-9 levels in saliva and GCF reflect inflammatory burden during pregnancy. They could be used for monitoring the inflammatory state of gingival tissues during pregnancy. [ABSTRACT FROM AUTHOR]

• GCF aMMP-8 chairside test sensitivity was 83.9 % and 79.2 % for specificity. • Stage IV periodontitis group had higher GCF aMMP-8 levels than the other groups. • Salivary aMMP-8 levels of P-Stage IV, III and gingivitis groups were higher than healthy group. • Low aMMP-8 levels but not total/latent MMP-8 clearly indicates periodontal health. • aMMP-8 is useful diagnostic biomarker for early detection in the new periodontitis classification. There is a need for a reliable complementary diagnostic tool that ideally helps to screen, differentiate sites, activities of and predict future periodontal tissue destruction. The purpose of this cross-sectional study was to investigate the screening and prevention potential of the chair-side/point-of-care (PoC) diagnostic test of salivary active matrix metalloproteinase-8 (aMMP-8) levels at different stages of periodontal disease and periodontal health. 80 individuals were included in this study; 18 with periodontitis stage 3 (P-Stage III), 19 with periodontitis stage-4 (P-Stage IV), 21 with gingivitis, and 22 with clinically healthy periodontium (H). The aMMP-8 levels in GCF and saliva were analyzed by chairside point-of-care aMMP-8 lateral flow immunotest and also by a time-resolved immunofluorescence assay (IFMA). The sensitivity of the chair-side/PoC test was 83.9 % while specificity was 79.2 %. The aMMP-8 IFMA levels in GCF were significantly higher in P-Stage IV group than P-Stage III, gingivitis and healthy groups (p = 0.01, p = 0.001, p = 0.00, respectively). Moreover, P-Stage III and gingivitis groups had significantly higher aMMP-8 IFMA levels than the healthy group (p < 0.05). The aMMP-8 chair-side test showed promising results in its ability to recognize and predict the inflammatory status even at the very initial/early stages. aMMP-8 chair-side test could be a valuable adjunctive diagnostic and preventive tool to conventional clinical methods in detecting periodontal disease. [ABSTRACT FROM AUTHOR]

This study investigated the diagnostic utility of mouthrinse and saliva in aMMP-8 measurements to analyze patients' risk for active periodontal tissue destruction and progression of periodontal disease among 47 adolescents. Results show that measurements from mouthrinse produce better discrimination and should be used instead of saliva measurements. [ABSTRACT FROM AUTHOR]