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Figure 2. Molecular forms and expression of MMP-1 and MMP-8 in GCF from the orthodontic and control patients detected by the Western blot method. (A) Western blot with MMP-8 specific antibody. Positions of molecular-weight markers (kDa) are indicated on the left. GCF samples were collected from the teeth undergoing orthodontic tooth movement immediately before (0 hr) and at 1-8 hrs after activation of fixed appliances. MMP-8 expression is undetectable before orthodontic force (0 hr); however, it is markedly increased after force application (1-8 hrs). PMN indicates culture supernatant of human neutrophils used as positive control for MMP-8. Bands in the range 75 to 80 kDa corresponding to PMN-type pro-MMP-8 are present at 1-8 hrs, whereas PMN active enzyme (60 kDa) is present at 4 hrs and at 7-8 hrs. Low-molecular-weight staining at < 30 kDa, representing degraded fragments of MMP-8, is present at 6-8 hrs. (B) Barely detectable MMP-8 immunoreactivity is seen in GCF control samples. (C) No MMP-1 immunoreactivity is detectable in the Western blots of the orthodontic GCF with MMP-1 specific antibody. As for positive control, MMP-1 from human rheumatoid synovial fibroblasts was used (F). (D) GCF control samples show no MMP-1 immunoreactivity.
