In vitro expression of matrix metalloproteinase-1, tissue inhibitor of metalloproteinase-1 and transforming growth factor-beta 1 in human periodontal ligament fibroblasts

Abstract

Extracellular matrix remodelling is mediated via matrix metalloproteinases (MMPs) and their regulatory factors such as tissue inhibitors of metalloproteinases (TIMPs) and transforming growth factor-ss (TGF-ss). 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-ss 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 Scheffe'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-ss1 expression and a dose-dependent increase in TIMP-1 and TGF-ss1 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-ss1 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.