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Objective: To determine whether oral rinse matrix metalloproteinase (MMP)-8 levels, measured by three different methods, tissue inhibitor of matrix metalloprotease-1 (TIMP-1) levels and elastase activity differentiate subjects with different periodontal condition; and second, to find out if MMP-8 levels were comparable among the methods used., Methods: MMP-8 levels were analysed with an immunofluorometric method (IFMA), dentoELISA and commercial ELISA. Also TIMP-1 levels and elastase activity were measured. For statistical analysis 214 study subjects were categorized into four groups, specified by the presence and number of moderate (4-5mm) and deep (≥6mm) periodontal pockets, and bleeding on probing percentage., Results: MMP-8 levels especially measured by dentoELISA and adjusted to the number of teeth per subject differentiated the study group with strong periodontal inflammatory burden from groups with lower levels. This was also verified with receiver operating characteristic (ROC) analysis. Elastase activity associated with higher IFMA and dentoELISA MMP-8 levels. IFMA MMP-8/TIMP and dentoELISA MMP-8/TIMP-1 tended to be higher with the increasing level of periodontal inflammatory burden. TIMP-1 levels decreased with increasing age., Conclusions: Oral rinse MMP-8 together with TIMP-1 analysis may have potential in complementary periodontal diagnostics. dentoELISA can be applied in quantitative oral rinse chair side biomarker diagnostics., (© 2010 John Wiley & Sons A/S.)

Objective: The aim of the study was to compare four methods for gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 detection., Methods: 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., Results: 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., Conclusions: 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.

The objective of this investigation was to determine the microbial generation time during early human supragingival plaque formation. On 2 occasions, 13 participants refrained from all oral hygiene measures for periods of 1, 2, 4, 8, 24 and 72 h. Plaque was sampled from defined areas of the teeth 13, 14, 15, 23, 24 and 25. Two independent methods were used to estimate the total number of viable cells: colony-forming units BCVF, total bacterial counts (BC) as enumerated by microscopic examination, and the percentage of vital bacteria using a vital fluorescence technique (VF) were recorded. The total number of vital microorganisms was calculated by BCVF (= BC x VF). The generation time based on either colony-forming units or BCVF increased significantly with plaque age. During the first interval (1-4 h), the microbial multiplication took slightly less than 1 h. After the establishment of a more mature plaque (24-72 h), the replicating time was between 12.5 h (BCVF) and 14.8 h (colony-forming units). It was concluded that the microbial generation time recorded at the initial stages of plaque formation is considerably lower than that found in older plaque.