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

• Published in: Japanese journal of ophthalmology Vol. 52; no. 1; pp. 8 - 15
• Main Authors: Yamanaka, Osamu, Saika, Shizuya, Ikeda, Kazuo, Miyazaki, Ken-ichi, Kitano, Ai, Ohnishi, Yoshitaka
• Format: Journal Article
• Language: English
• Published: Japan Springer Japan 01.02.2008; Springer Nature B.V
• Subjects: Cell Movement - physiology; Cell Proliferation; Cells, Cultured; Child; Child, Preschool; Collagen Type I - genetics; Collagen Type I - metabolism; Conjunctiva - cytology; Conjunctiva - drug effects; Conjunctiva - metabolism; Connective Tissue Growth Factor; DNA, Antisense - pharmacology; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix - drug effects; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Fibronectins - metabolism; Humans; Immediate-Early Proteins - physiology; Intercellular Signaling Peptides and Proteins - physiology; Laboratory Investigation; Medicine; Medicine & Public Health; Oligonucleotides, Antisense - pharmacology; Ophthalmology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Transforming Growth Factor beta1 - pharmacology; connective tissue growth factor; wound healing; filtration surgery; transforming growth factor β; extracellular matrix
• ISSN: 0021-5155; 1613-2246
• DOI: 10.1007/s10384-007-0497-3

Purpose We examined the role of connective tissue growth factor (CTGF) in transforming growth factor β1 (TGFβ1)-related behavior in cultured human subconjunctival fibroblasts (SCFs), protein production, mRNA expression of CTGF and type I collagen α1 chain (colIA1), and cell proliferation and migration. TGFβ1 is the major factor involved in bleb scarring following filtration surgery. Methods An antisense deoxynucleotide (antisense) (5 μM) 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 TGFβ1 (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 TGFβ1 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 TGFβ1 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 TGFβ1 in vitro. Inhibition of CTGF expression may be effective in preventing undesirable scar formation during healing following filtration surgery.