Connective tissue growth factor modulates extracellular matrix production in human subconjunctival fibroblasts and their proliferation and migration in vitro

Purpose: We examined the role of connective tissue growth factor (CTGF) in transforming growth factor beta 1 (TGF beta 1)-related behavior in cultured human subconjunctival fibroblasts (SCFs), protein production, mRNA expression of CTGF and type I collagen alpha 1 chain (colIA1), and cell proliferation and migration. TGF beta 1 is the major factor involved in bleb scarring following filtration surgery. Methods: An antisense deoxynucleotide (antisense) (5 mu M) for CTGF mRNA was used to block endogenous CTGF expression. Effects of antisense on extracellular matrix (ECM) production and immunolocalization, mRNA expression, and cell proliferation and migration were examined in human SCF cultures with or without TGF beta 1 (5 ng/ml). Cell migration was examined in an in vitro wound model of monolayer fibroblast cultures. Results: CTGF antisense reduced mRNA expression of CTGF and colIA1 and production of the ECM components type I collagen, and fibronectin much more markedly in cells treated with TGF beta 1 compared with control fibroblasts, and it inhibited the proliferation of cultured SCFs to 71.9% of that of controls after 13 days of culture. CTGF antisense also delayed defect closure in monolayer cell sheets. In the culture, the defect was closed by TGF beta 1 by 36 h, whereas 7.0% of the defect remained at 48 h in the antisense-treated culture. Conclusions: These findings indicate that CTGF is involved in ECM production in SCFs activated by exogenous TGF beta 1 in vitro. Inhibition of CTGF expression may be effective in preventing undesirable scar formation during healing following filtration surgery.