Gingival Crevicular Fluid MMP-8 Levels Research Articles

Dynamics of Matrix Metalloproteinase-1 and -8 Secretion in Gingival Crevicular Fluid after Gingival Recession Therapy via MCAT with Either Subepithelial Connective Tissue Graft or Collagen Matrix.

Objectives: The objective of this study was to determine and estimate the changing levels of matrix metalloproteinases 1 and 8 (MMP-1 and MMP-8) in GCF at consecutive stages of healing after root coverage procedure via modified coronally advanced tunnel (MCAT) combined with either sub-epithelial connective tissue graft (SCTG) or collagen matrix (CM) and also to relate those changes to clinical outcomes of both therapeutic approaches. Materials and methods: The study involved 20 patients with a total of 91 recessions. Those on one side of the mandible received MCAT plus CM while the contralateral ones MCAT plus SCTG. The evaluation of MMP-1 and MMP-8 concentrations in Gingival Crevicular Fluid (GCF) took place at baseline, then at 1, 2, and 4 weeks, and finally at 3 months after surgery. Elisa protocol was applied to determine the levels of MMP-1 and MMP-8 in GCF. Results: Three-month observation revealed statistically significant changes in MMP-1, MMP-8 and Sulcus Fluid Flow Rate (SFFR) values after implementation of both techniques. A correlation was found between a difference in MMP-1 concentrations and gain in Keratinized Tissue (KT) after SCTG and CM. MMP-8 levels and a Gingival Thickness (GT) gain observed after CM was also correlated. Conclusions: A type of augmentative material does appear to determine the dynamics of MMP-1 secretion.

Gingival Crevicular Fluid Zinc- and Aspartyl-Binding Protease Profile of Individuals with Moderate/Severe Atopic Dermatitis.

Atopic dermatitis (AD) is a protease-modulated chronic disorder with heterogenous clinical manifestations which may lead to an imprecise diagnosis. To date, there are no diagnostic protease tests for AD. We explored the gingival crevicular fluid (GCF) protease profile of individuals with moderate/severe AD compared to healthy controls. An exploratory case-control study was conducted. AD patients (n = 23) and controls (n = 21) were enrolled at the International Center for Clinical Studies, Santiago, Chile. Complete dermatological and periodontal evaluations (involving the collection of GCF samples) were made. The levels of 35 proteases were analyzed using a human protease antibody array in matching AD patients (n = 6) and controls (n = 6) with healthy periodontium. The GCF levels of zinc-binding ADAM8, ADAM9, MMP8, Neprilysin/CD10, aspartyl-binding Cathepsin E, serin-binding Protein convertase9, and uPA/Urokinase proteases were lower in moderate/severe AD patients compared to controls (p < 0.05). No inter-group differences in the levels of the other 28 proteases were found. MMP8, Cathepsin E, and ADAM9 were the biomarkers with the highest sensitivity and specificity regarding the detection of AD (p < 0.05). The area under receiver operating characteristic (ROC) curve for MMP8 was 0.83 and MMP8 + ADAMP9 was 0.90, with no significant differences (p = 0.132). A combined model of MMP8, Cathepsin E, and ADAM9 was not considered since it did not converge. Then, levels of MMP8 in GCF were determined using a multiplex bead immunoassay in 23 subjects with AD and 21 healthy subjects. Lower levels of MMP8 in the GCF from the AD group versus healthy group (p = 0.029) were found. This difference remained significant after adjustment by periodontitis (p = 0.042). MMP8 revealed the diagnostic potential to identify AD patients versus healthy controls, (ROC area = 0.672, p < 0.05). In conclusion, differences in the protease profile between AD and control patients were associated with MMP8, Cathepsin E, and ADAM9. Based on the multiplex assay results, MMP8 was lower in AD patients than controls, suggesting that MMP8 may be a diagnostic biomarker candidate.

Importance of MMP-8 in Salivary and Gingival Crevicular Fluids of Periodontitis Patients.

Matrix metalloproteinases (MMPs) stimulate alveolar bone loss in chronic periodontitis. To evaluate the salivary and gingival crevicular fluid (GCF) levels of MMP-8 in patients with moderate to severe chronic periodontitis. 42 participants were divided into two groups: a case group (21 patients with generalized moderate to severe chronic periodontitis) and a control group (21 healthy periodontal subjects). GCF and saliva samples were obtained from both groups. Salivary and GCF MMP-8 levels of each subject were detected using the ELISA method. Mean±SD values of salivary MMP-8 levels of the control and case groups were 1.52 ± 0.65 ng/ml and 6.06 ± 1.18 ng/ml, respectively, and statistically significant difference was observed (p=0.0001). Also, mean±SD values of GCF MMP-8 levels of the control and case groups were 0.87 ± 0.26 ng/ml and 2.92 ± 0.64 ng/ml, respectively; which was statistically significant (p=0.0001). Our results demonstrate an increased concentration of salivary and GCF levels of MMP-8 in the patient group.

Evaluation of the clinical and biochemical efficacy of erbium, chromium:ytrium-scandium-gallium-garnet (ER,CR:YSGG) laser treatment in periodontitis.

The objective of this study was to evaluate the clinical and biochemical efficacy of erbium, chromium:ytrium-scandium-gallium-garnet (Er,Cr:YSGG) laser treatment, in addition to non-surgical periodontal treatment (NSPT), in periodontitis patients. This was a single-centre, split-mouth, randomized, controlled 6-month clinical trial. Twenty-seven patients with moderate to advanced periodontitis were included in the study. The patients were randomly assigned to a debridement and laser treatment group or control group (debridement alone). Clinical measurements were performed at baseline and 1, 3 and 6months post-treatment. The following parameters were measured: plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL), bleeding on probing (BoP), gingival crevicular fluid (GCF) volume and IL-1β and MMP-8 levels in GCF. There were statistically significant clinical improvements in both groups, with no significant between-group differences in PD, CAL, PI and BoP clinical measurements (p < 0.05). As compared with the control group, the GI 1 and 3months post-treatment and BoP 1month post-treatment were significantly improved in the laser group (p < 0.05). There was also a significant reduction in IL-1 β levels in both groups at baseline versus those 1, 3 and 6months later (p < 0.05) There was no statistically significant difference in IL-1β and MMP-8 levels between groups (p < 0.05). Both the treatment modalities resulted in significant improvements in clinical parameters. Within the limitations of this study, Er,Cr:YSGG laser treatment applied in addition to NSPT is indicated to reduce clinical inflammation.

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.

Correlations of inflammatory cytokines, oxidative stress markers, and matrix metalloproteinases in gingival crevicular fluid with peri-implantitis

This study aimed to analyze the correlations of inflammatory cytokines, oxidative stress markers, and matrix metalloproteinases (MMPs) in gingival crevicular fluid (GCF) with peri-implantitis (PI). Forty patients receiving dental implantation were enrolled. There were 52 implants, which were divided into PI group (42 implants) and health implant (HI) group (10 implants). Fifty-two healthy teeth (HT) with the same names with affected teeth in the patients were selected as the control group. The periodontal status was recorded. The GCF was collected and quantified. The levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), hypersensitive C-reactive protein (hs-CRP), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), MMP-13, and MMP-8 were detected using enzyme-linked immunosorbent assay (ELISA). Results showed that the probing depth, sulcus bleeding index, GCF volume, and TNF-α, IL-6, hs-CRP, MMP-8, and MMP-13 levels in GCF in PI group were significantly higher than HI and HT groups, respectively ( P &lt; 0.01 or P &lt; 0.05). The SOD and GSH-Px levels in PI group were significantly lower than HI and HT groups, respectively ( P &lt; 0.05). Excepting hs-CRP, there was no significant difference of each index between HI and HT groups ( P &gt; 0.05). In conclusion, TNF-α, IL-6, hs-CRP, SOD, GSH-Px, MMP-8, and MMP-13 are involved in the occurrence of PI, and they may be used as auxiliary indicators to evaluate the degree of PI. In addition, the clinical periodontal index probing depth and sulcus bleeding index are positively correlated with GCF volume, hs-CRP, MMP-8, and MMP-13.

Associations of oral fluid MMP-8 with periodontitis in Swiss adult subjects.

MMP-8 is a prominent collagenase in periodontal disease. This cross-sectional study examined whether MMP-8 levels in saliva and gingival crevicular fluid (GCF) are associated with periodontitis in a Swiss population. A total of 258 subjects (107m, 151 f, mean age: 43.5yr; range: 21-58yr) acquired from the Swiss bone marrow donor registry participated in the study. Saliva and GCF samples were collected from subjects followed by a thorough dental and periodontal examination. MMP-8 levels were determined with immunofluorometric assay. Associations of MMP-8 levels with periodontal diagnosis, probing pocket depth (PPD) and bleeding on probing were statistically analysed with Pearson chi-square test, Spearman's rho and logistic regression analysis. MMP-8 in GCF correlated with MMP-8 in saliva (p<.001). Periodontitis was more common (p<.001) among subjects with high levels of MMP-8 in saliva and/or GCF compared with subjects with low levels of MMP-8. Higher MMP-8 levels in GCF and saliva were associated with any periodontal diagnosis (mild, moderate or severe), greater PPD, and bleeding on probing (p<.05). When age, gender, smoking, body mass index, number of medications and decayed, missing and filled teeth were adjusted for, all observed associations remained statistically significant. The area under curve of receiver-operating characteristic was 0.67 for saliva and 0.71 for GCF. Elevated MMP-8 levels both in saliva and GCF are associated with periodontitis in a normal adult population.

Concentration of MMP-8 and IL-1β in gingival crevicular fluid in patients with chronic and aggressive periodontitis.

Modern research confirms the role of inflammatory mediators in the pathomechanism of periodontal tissue destruction. The aim of the study was to determine concentrations of MMP-8 and IL-1β in gingival crevicular fluid (GCF) in patients with advanced chronic and aggressive periodontitis. The authors measured the concentrations of the above inflammatory mediators in gingival crevicular fluid of deep pockets (PD ≥ 6 mm) and shallow pockets (PD 4-5 mm) in 33 patients with advanced chronic periodontitis and in 16 patients with aggressive periodontitis. The control group consisted of 16 individuals with healthy periodontium. In all patients levels of MMP-8 and IL-1β in GCF were determined with the ELISA method. The study showed significantly higher concentrations of MMP-8 and IL-1β in GCF of both deep and shallow pockets in patients with periodontitis compared to healthy subjects. No difference in concentrations of the tested mediators was observed with reference to diagnosis of aggressive periodontitis (AP) or chronic periodontitis (CP).

The utility of gingival crevicular fluid matrix metalloproteinase-8 response patterns in prediction of site-level clinical treatment outcome.

Different gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 response patterns were studied among non-smoking and smoking patients with chronic periodontitis (CP) and generalized aggressive periodontitis (GAgP) to test the utility of GCF MMP-8 levels predicting the site-level treatment outcome. Data from four independent longitudinal studies were combined. Altogether, the studies included 158 periodontal sites from 67 patients with CP and 32 patients with GAgP, and GCF samples were collected at baseline, after the treatment, and during the 6-month maintenance period. All GCF samples were analyzed by immunofluorometric assay for MMP-8. Different site-level MMP-8 response patterns were explored by the cluster analysis. Most optimal MMP-8 cutoff levels were searched with receiver operating characteristic analyses, and the predictive utility of defined levels was tested. Distinct types of MMP-8 response patterns were found in both smokers and non-smokers. MMP-8 levels exceeding the optimal cutoff levels separately defined for smokers and non-smokers indicated increased risk for compromised treatment outcome at baseline and during the maintenance period. Seventy-one percent of non-smokers (positive likelihood ratio of 4.22) and 88% of smokers (positive likelihood ratio of 5.00) with positive test results at both baseline and the maintenance period had compromised treatment outcome. The double-positive result indicated 46% and 39% point risk increase for the compromised outcome, respectively. GCF MMP-8 analysis with defined cutoff levels could be used to predict the site-level treatment outcome and for longitudinal monitoring of the disease status during the maintenance period.

Host-derived biomarkers at teeth and implants in partially edentulous patients. A 10-year retrospective study.

To assess a selection of host-derived biomarkers in peri-implant sulcus fluid (PISF) and gingival crevicular fluid (GCF) from adjacent teeth 10years following implant placement. Peri-implant sulcus fluid and GCF samples obtained from the deepest sites of 504 implants and 493 adjacent teeth were analysed for levels of interleukin (IL)-1β, matrix metalloproteinase (MMP)-3, MMP-8, MMP-1, and MMP-1 bound to tissue inhibitor of MMP (TIMP)-1 (MMP-1/TIMP-1) by enzyme-linked immunosorbent assay (ELISA) technique. Overall, MMP-8 was detected in 90% of the sites. In more than 50% of the sites, IL-1β was identified while in 30% of the sites MMP-1, MMP-1/TIMP-1 and MMP-3 were found over the detection level. Increased biomarkers levels from PISF and GCF were positively correlated (r=0.375-0.702; P<0.001). However, no qualitative and quantitative differences were found between PISF and GCF. The levels of MMP-1 were negatively correlated with those of MMP-1/TIMP-1 at implants (r=-0.644; P<0.001). Median MMP-1 levels at implants were high (5.17pg/site) in subjects with severe chronic periodontitis and low in patients with mild-to-moderate chronic periodontitis (0pg/site; P=0.026) or gingivitis (0pg/site; P=0.034). Levels of IL-1β were found to be different in GCF according to the periodontal conditions (P=0.001) with the highest level found in mild-to-moderate periodontitis (6.2pg/site). Clinical attachment levels at implants demonstrated an inverse correlation with MMP-1/TIMP-1 (r=-0.147; P=0.001). Increased levels of MMP-8 and IL-1β in PISF or GCF may be associated with inflammation around teeth and implants while lower levels of MMP-1/TIMP-1 may be an indicator of disease progression around implants.

Evaluation of GCF MMP-1, MMP-8, TGF-β1, PDGF-AB, and VEGF levels in periodontally healthy smokers

The effect of smoking on inflammatory biomarkers in gingival crevicular fluid (GCF) is well established in the presence of periodontal inflammation. However, it is not clear if smoking has an influence on matrix metalloproteinase (MMP) and growth factor levels in the GCF of periodontally healthy subjects. The aim of this study was to investigate GCF levels of MMP-1, MMP-8, transforming growth factor (TGF)-β1, platelet-derived growth factor (PDGF)-AB, and vascular endothelial growth factor (VEGF) in smoking versus nonsmoking periodontally healthy subjects. Thirty-two periodontally healthy subjects were included in this study. Probing depths, bleeding on probing, and plaque index was assessed. GCF levels of MMP-1, MMP-8, TGF-β1, PDGF-AB, and VEGF were analyzed by enzyme-linked immunosorbent assay. No significant differences were observed in the distribution of demographic data between study groups. GCF total amount of PDGF-AB was significantly lower in smokers compared to nonsmokers (P = 0.014). Total amount of GCF MMP-1, MMP-8, TGF-β1, and VEGF levels were similar in both study groups (P = 0.022). Smoking has the effect of decreasing GCF PDGF-AB while it does not affect GCF MMP-1, MMP-8, TGF-β1, and VEGF in periodontally healthy subjects. Since increased levels of these molecules are involved in periodontal breakdown, our findings may emphasize the importance for maintenance of periodontal health in smokers.

Analysis of matrix metalloproteinase-8 levels in gingival crevicular fluid and whole mouth fluid among smokers and nonsmokers using enzyme-linked immune-sorbent assay and a novel chair-side test.

Aim:To indigenously prepare a chair-side test kit for investigating and comparing the matrix metalloproteinase (MMP)-8 levels in gingival crevicular fluid (GCF) and saliva in patients with healthy periodontium, gingivitis and chronic periodontitis in smokers and nonsmokers. To validate the diagnostic accuracy of indigenously prepared chair-side test against enzyme-linked immune-sorbent assay (ELISA). Furthermore, to assess the effect of nonsurgical periodontal therapy (NSPT) on the levels of MMP-8 in GCF and saliva among the test groups.Materials and Methods:GCF and saliva were collected from 250 subjects. The study population were divided into five groups; health periodontium-nonsmokers (Group 1; n = 50), chronic gingivitis-nonsmokers (Group 2; n = 50), chronic periodontitis-nonsmokers (Group 3; n = 50), chronic gingivitis-smokers (Group 4; n = 50), chronic periodontitis-smokers (Group 5; n = 50). A chair-side test kit was indigenously prepared using polyclonal antibodies (principle of immunochromatography) to detect the MMP-8 levels, and it was validated against ELISA at baseline and 3 months after NSPT.Results:The chair-side test detected MMP-8 levels with a sensitivity and specificity in accordance with ELISA. MMP-8 levels at baseline were higher in Group 2 and Group 3 as compared to controls (P < 0.05), and decreased after therapy (P < 0.05). MMP-8 levels in GCF were greater than in saliva for all the groups, indicating GCF to be a better sample to detect the MMP levels.Conclusion:The chair-side test detected MMP-8 levels accurately making it a viable chair side diagnostic tool. It was effective for early diagnosis of the periodontal disease among high-risk population such as smokers.

Matrix Metalloproteinase (MMP)‐8 and Tissue Inhibitor of MMP‐1 (TIMP‐1) Gene Polymorphisms in Generalized Aggressive Periodontitis: Gingival Crevicular Fluid MMP‐8 and TIMP‐1 Levels and Outcome of Periodontal Therapy

The aim of the present study is to investigate matrix metalloproteinase (MMP)-8 and tissue inhibitor of MMP-1 (TIMP-1) gene polymorphisms in generalized aggressive periodontitis (GAgP) and to assess the effects of MMP-8 and TIMP-1 genotypes on the outcomes of non-surgical periodontal therapy. Genomic DNA was obtained from peripheral blood of 100 patients with GAgP and 167 periodontally healthy controls. MMP-8 +17 C/G, -799 C/T, -381 A/G and TIMP-1 372 T/C, *429 T/G polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism. Patients with GAgP received non-surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and gingival crevicular fluid (GCF) samples were collected at baseline and at follow-up visits. GCF biomarkers were analyzed by immunofluorescence assay and enzyme-linked immunosorbent assay. Distribution of the MMP-8 -799 C/T genotypes was significantly different between the GAgP and control groups (P <0.005). TIMP-1 372 T/C and *429 T/G genotypes in males were also significantly different between study groups (P <0.004). GCF MMP-8 levels decreased until 3 months after non-surgical therapy compared with baseline in T and G alleles, as well as G and C allele carriers (P <0.0125), whereas no significant decreased was observed in non-carriers (P >0.0125). On the basis of the present findings, it can be suggested that MMP-8 -799 C/T and TIMP-1 372 T/C, *429 T/G gene polymorphisms in males may be associated with the susceptibility to GAgP in the Turkish population.

Effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis

Emingil G, Han B, Özdemir G, Tervahartiala T, Vural C, Atilla G, Baylas H, Sorsa T. The effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis. J Periodont Res 2012; 47: 729-739. © 2012 John Wiley & Sons A/S Background and Objective: To study the effectiveness of azithromycin in combination with nonsurgical periodontal therapy on clinical and microbiological parameters, and on the MMP-8 and TIMP-1 levels in gingival crevicular fluid, over a 6-mo time-period in patients with generalized aggressive periodontitis. Thirty-two patients with generalized aggressive periodontitis were included in this randomized, double-blind, placebo-controlled, parallel-arm study. They were randomly assigned to azithromycin or placebo groups (500 mg once daily for 3 d). Probing depth, clinical attachment levels, presence of bleeding on probing and plaque were recorded. Gingival crevicular fluid samples were obtained from one single-rooted tooth, while microbiological samples were obtained from two single-rooted teeth, all with a probing depth of ≥ 6 mm. Microbiological parameters were analyzed by quantitative real-time PCR for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and total bacteria. Gingival crevicular fluid biomarkers were determined by immunofluorometric assay and ELISA. All clinical parameters improved, and microbiological parameters and gingival crevicular fluid MMP-8 levels significantly decreased, over the 6-mo period (p < 0.05); both groups demonstrated similar improvements. The azithromycin group presented a higher percentage of deep pockets resolved (probing depth reduction of ≥ 3 mm from baseline) compared with the placebo group at 1 mo (p < 0.05). Adjunctive azithromycin therapy provides no additional benefit over nonsurgical periodontal treatment on clinical parameters, microbiological parameters and gingival crevicular fluid biochemical markers investigated in patients with generalized aggressive periodontitis.

Gingival crevicular fluid MMP-8-concentrations in patients after acute myocardial infarction

BackgroundThe aim of this study was to determine the presence of matrix metalloproteinase-8 in the gingival crevicular fluid (GCF) of patients after acute myocardial infarction (AMI).MethodsA total of 48 GCF samples from 20 AMI patients, hospitalized at the Department of Cardiology and Angiology of the Johannes Gutenberg University Mainz, were investigated. Besides the myocardial infarction all patients suffered from chronic periodontal disease. Fifty-one GCF samples from 20 healthy age matched individuals with similar periodontal conditions served as controls. The dental examination included the assessment of oral hygiene, gingival inflammation, probing pocket depth, clinical attachment level and X-ray examination. The study was only carried out after the positive consent of the regional ethic commission. A quantitative assessment of aMMP-8 levels in the gingival crevicular fluid was performed with the help of the DentoAnalyzer (Dentognostics GmbH, Jena, Germany), utilising an immunological procedure.ResultsThe aMMP-8 concentrations found in the gingival crevicular fluid of the AMI patients significantly differed (p = 0.001; mean value 30.33 ± 41.99 ng/ml aMMP-8) from the control group (mean value 10.0 ± 10.7 ng/ml aMMP-8). These findings suggest that periodontal inflammation in AMI patients might be associated with higher MMP-8-values compared to the healthy controls.ConclusionsThe acute myocardial infarction seems to influence the degree of periodontal inflammation, thus the measurement of the gingival crevicular fluid MMP8 levels seems to be a helpful biochemical test to obtain information about the severity of the periodontal disease.

Gingival Crevicular Fluid MMP‐8 and −13 and TIMP‐1 Levels in Patients With Rheumatoid Arthritis and Inflammatory Periodontal Disease

The purpose of this study was to compare gingival crevicular fluid (GCF) levels of matrix metalloproteinase (MMP)-8 and -13 and tissue inhibitor of MMP (TIMP)-1 in patients with rheumatoid arthritis (RA) and systemically healthy counterparts with inflammatory periodontal disease. Subjects (N = 74) were divided into five groups: 12 patients with RA and gingivitis; 13 patients with RA and periodontitis; 12 systemically healthy patients with gingivitis; 13 systemically healthy patients with periodontitis; and 24 periodontally and systemically healthy volunteers. Full-mouth clinical periodontal measurements were performed at six sites/tooth. GCF samples obtained from two sites in single-rooted teeth were analyzed by immunofluorometric assay and enzyme-linked immunosorbent assay. Data were assessed statistically by parametric tests. The total amounts of MMP-8 were lower in the healthy control group than in RA-gingivitis, RA-periodontitis, and healthy-periodontitis groups (P <0.05). MMP-13 levels were similar in all five study groups (P >0.05). Patients with RA and gingivitis or periodontitis exhibited levels of MMP-8 and -13 and TIMP-1 that were similar to systemically healthy counterparts (P >0.05). The coexistence of RA and periodontitis did not significantly affect the investigated parameters. GCF MMP-8 levels increased with periodontal inflammation. Despite the long-term usage of corticosteroids and non-steroidal anti-inflammatory drugs, similar GCF MMP-8 and -13 levels in patients with RA and systemically healthy counterparts suggest that RA may create a tendency to overproduce these enzymes.

Detection of gingival crevicular fluid MMP-8 levels with different laboratory and chair-side methods

The aim of the study was to compare four methods for gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 detection. Matrix metalloproteinase-8 levels from 20 GCF samples from two periodontally healthy subjects, 18 samples from two patients with gingivitis and 45 samples from six patients with moderate to severe periodontitis, altogether 83 samples, were analysed using (1) a time-resolved immunofluorometric assay (IFMA), (2) an MMP-8 specific chair-side dip-stick test, (3) a dentoAnalyzer device and (4) the Amersham ELISA kit. Western immunoblot using same monoclonal anti-MMP-8 as in IFMA and dentoAnalyzer was used to identify molecular forms of MMP-8 in GCFs. Correlation between IFMA and dentoAnalyzer results calculated with Spearman's correlation coefficient was 0.95 (P = 0.01). The chair-side dip-stick test results were well in line with these assays. Periodontitis sites with unstable characteristics were differentiated with these methods. The Amersham ELISA results were not in line with the findings by other methods. Immunofluorometric assay and dentoAnalyzer can detect MMP-8 from GCF samples and these methods are comparable. Using Western immunoblot, it was confirmed that IFMA and dentoAnalyzer can detect activated 55 kDa MMP-8 species especially in periodontitis-affected GCF. dentoAnalyzer is among the first quantitative MMP-8 chair-side testing devices in periodontal and peri-implant diagnostics and research.

GCF MMP‐8 Levels in Smokers and Non‐Smokers With Chronic Periodontitis Following Scaling and Root Planing Accompanied by Systemic Use of Flurbiprofen

Cigarette smoking has been identified as an important risk factor for the initiation and progression of chronic periodontitis (CP). The aim of this study was to investigate the effects of phase I periodontal therapy and adjunctive flurbiprofen administration on matrix metalloproteinase (MMP)-8 levels in gingival crevicular fluid (GCF) samples from smoking and non-smoking patients with CP. Twenty-nine non-smoking and 29 smoking patients with CP were divided into four groups according to periodontal treatment modalities. Group 1 (non-smokers with CP) and group 3 (smokers with CP) patients received daily 100-mg flurbiprofen tablets in a 2 x 1 regimen for 10 days together with scaling and root planing (SRP). Patients in group 2 (non-smokers with CP) and group 4 (smokers with CP) received placebo tablets in a 2 x 1 regimen for 10 days together with SRP. Plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) measurements were recorded; GCF samples were collected from each sampling area at baseline and after the 10-day period of drug intake by a single examiner who was unaware of the treatment modality. Assays for GCF MMP-8 were carried out by an enzyme-linked immunosorbent assay. All groups showed statistically significant reductions in PI and GI scores following the phase I periodontal treatment (P < 0.05), but no statistical differences were observed in PD and CAL scores after therapy. In all groups, the reduction of GCF MMP-8 levels after therapy was statistically significant compared to baseline levels (P < 0.001). When groups 1 and 3 and 2 and 4 were compared according to GCF MMP-8 levels after the therapy, no statistically significant differences were observed (P = 0.117 and P = 0.485, respectively). Flurbiprofen administration had no additional inhibitory effect over SRP alone on GCF levels of MMP-8 in smokers compared to non-smokers with CP.

Monitoring periodontal disease status in smokers and nonsmokers using a gingival crevicular fluid matrix metalloproteinase‐8‐specific chair‐side test

With current periodontal diagnostic tools it is difficult to identify susceptible individuals or sites at risk. The aim of this study was to evaluate the efficacy of the matrix metalloproteinase (MMP)-8-specific chair-side dip-stick test in longitudinally monitoring the periodontal status of smoking (S) and nonsmoking (NS) patients with chronic periodontitis, using their gingival crevicular fluid (GCF) MMP-8 concentrations. Clinical parameters, MMP-8 test results and concentrations were monitored in 16 patients after initial treatment and in 15 patients after scaling and root planing (SRP), every other month, over a 12-mo time period. Progressing and stable sites, and sites with exceptionally high MMP-8 concentrations, were analysed in smokers and nonsmokers. SRP reduced the mean GCF MMP-8 levels, test scores, probing depth (PD), attachment loss (AL) and bleeding on probing (BOP). In sites of periodontal disease progression, the distribution of MMP-8 concentrations was broader than in stable sites, indicating a tendency for elevated concentrations in patients with periodontal disease. The mean MMP-8 concentrations in smokers were lower than in nonsmokers, but in smokers' and nonsmokers' sites with progressive disease, MMP-8 concentrations were similar. Sites with exceptionally elevated MMP-8 concentrations were clustered in smokers who also showed a poor response to SRP. In these sites, the MMP-8 concentration did not decrease with SRP and these sites were easily identified by the MMP-8 test. Persistently elevated GCF MMP-8 concentrations may indicate sites at risk, as well as patients with poor response to conventional periodontal treatment (e.g. SRP). MMP-8 testing may be useful as an adjunct to traditional periodontal diagnostic methods during the maintenance phase.

The effect of topical doxycycline usage on gingival crevicular fluid MMP‐8 levels of chronic and aggressive periodontitis patients: a pilot study

The aim of this study was to evaluate the efficacy of topical subgingival application of doxycycline hyclate (DH) gel adjunctive to non-surgical periodontal therapy on gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 levels in chronic and aggressive periodontitis patients. Forty teeth of 10 chronic periodontitis patients and 32 teeth of eight aggressive periodontitis patients were screened for 6 months. Scaling and root planing (SRP) was applied to the control sites and DH gel adjunctive to SRP was applied to the test sites of each patient simultaneously. GCF MMP-8 levels were analysed at baseline, 7 days; and at 1, 3 and 6 months by Sandwich Elisa Method. At 1, 3 and 6 months, probing depth (P < 0.0051) and plaque scores and bleeding on probing values (P = 0.000) significantly decreased in each group when compared with the baseline, but there was no statistically significant difference between the test and control sites. GCF MMP-8 levels reduced presenting statistically significant differences on 7 days, 1, 3 and 6 months in four of the groups (P < 0.05); however, intergroup differences were not statistically significant. Developing functional and immunological-based chair-side MMP tests might serve as useful adjunctive diagnostic tools when monitoring the effects of DH gel application.