In vitro expression of matrix metalloproteinase-1, tissue inhibitor of metalloproteinase-1 and transforming growth factor-beta1 in human periodontal ligament fibroblasts.
Extracellular matrix remodelling is mediated via matrix metalloproteinases (MMPs) and their regulatory factors such as tissue inhibitors of metalloproteinases (TIMPs) and transforming growth factor-beta (TGF-beta). The regulation of MMPs is thought to be associated with cytoskeletal changes. In this study, cytoskeletal changes in human periodontal ligament fibroblasts (PDFs) were induced using cytochalasin B (CB) which reorganizes actin microfilaments reversibly, and colchicine which disrupts microtubules irreversibly. The levels of MMP-1, TIMP-1 and TGF-beta secreted by the CB- or colchicine-treated PDFs were measured using an enzyme-linked immunosorbent assay. Differences between experimental and control groups were tested using analysis of variance (ANOVA) and Scheffé's ad hoc test. Although CB treatment did not significantly increase MMP-1 expression over the controls, colchicine treatment significantly increased the expression of MMP-1 (P < 0.01) in a time-dependent manner compared with the controls. Both CB and colchicine showed a time-dependent increase in TIMP-1 and TGF-beta1 expression and a dose-dependent increase in TIMP-1 and TGF-beta1 expression until threshold compared with the controls (P < 0.05, P < 0.01 and P < 0.001). In addition, CB treatment produced significantly increased TGF-beta1 expression over the controls (P < 0.05 and P < 0.001) from lower doses, with this effect occurring at earlier time points compared with colchicine treatment.
Nahm
Dong-Seok
DS
Department of Orthodontics, College of Dentistry, Seoul National University, Seoul, Korea.
Kim
Hee-Jeong
HJ
Mah
James
J
Baek
Seung-Hak
SH
D008841:Actin Cytoskeleton / Q000187:drug effects; D000704:Analysis of Variance; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis*; D008870:Microtubules / Q000187:drug effects; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis*; D016212:Transforming Growth Factor beta / Q000096:biosynthesis*; D053773:Transforming Growth Factor beta1
C508960:TGFB1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D003571:Cytochalasin B; D020781:Matrix Metalloproteinase 1; D003078:Colchicine