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Cathepsin C, Matrix Metalloproteinases, and their Tissue Inhibitors in Gingiva and Gingival Crevicular Fluid from Periodontitis-affected Patients
M. Soell1,
R. Elkaim2 and
H. Tenenbaum1,*
1 Unité INSERM U 424, 11 rue Humann, 67085 Strasbourg CEDEX, France; and
2 Parogène, Strasbourg, France;
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Figure 1. Quantification of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, and TIMP-2 in tissue extract supernatants and gingival crevicular fluid samples from healthy and diseased patients. Evaluations were performed by means of a sandwich ELISA assay in tissue extract supernatants (TES/A) and gingival crevicular fluid samples (GCFs/B). Results were obtained from three different experiments (n = 3) and expressed as mean ± SE of µg of antigen/mg of total proteins. (*) indicated that the Mann-Whitney Rank Sum Test showed a statistically significant difference between diseased and healthy tissue extract supernatants (p < 0.003) and between diseased and healthy gingival crevicular fluid samples (p < 0.003).
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Figure 2. Proteolytic activities of MMPs in tissue extract supernatants and gingival crevicular fluid samples from healthy and diseased patients. The proteolytic activities of MMPs were determined in tissue extract supernatants (TES/A) and gingival crevicular fluid samples (GCFs/B) from healthy and diseased patients by quantification of the percentage of degradation of a biotinylated substrate: human collagen type-1 for MMP-1, casein for MMP-3, and gelatin for both MMP-2 and MMP-9. Controls in TES/A indicate the results obtained when 1 µg of each purified human MMP was used in the assay. Results were obtained from three different experiments (n = 3), and activity was expressed as mean ± SE percentage of biotinylated substrate degraded/mg of total proteins/hr. (*) indicated that the Mann-Whitney Rank Sum Test showed a statistically significant difference between diseased and healthy tissue extract supernatants (p < 0.002) and between diseased and healthy gingival crevicular fluid samples (p < 0.001).
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Figure 3. Proteolytic activities of cathepsin C in tissue extract supernatants and gingival crevicular fluid samples from healthy and diseased patients. Activity of cathepsin C in tissue extract supernatants (TES) and gingival crevicular fluid samples (GCFs) from healthy and diseased patients was determined by measurement of the amount of 7-amido-4 methyl-coumarin (AMC) released by hydrolysis of the synthetic substrate H-Gly-Arg-AMC. The AMC formed was monitored by means of a VersaFluor Multiple-Wavelength fluorometer luminescence spectrometer at 370-nm excitation and 460-nm emission. Results were obtained from three different experiments (n = 3) and expressed as mean ± SE of µMoles of AMC produced/min/mg of total proteins in the sample. (*) indicated that the Mann-Whitney Rank Sum Test showed a statistically significant difference between diseased and healthy tissue extract supernatants (p < 0.006) and between diseased and healthy gingival crevicular fluid samples (p < 0.003).
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Journal of Dental Research, Vol. 81, No. 3,
174-178 (2002)
DOI: 10.1177/154405910208100306
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