von Eicken J., Denk P. O.

Augenheilkunde I, University Eye Clinic, Tuebingen, Germany

Human Tenon's capsule fibroblasts (HTF) transdifferentiate to myofibroblasts during the wound healing response after glaucoma filtering surgery. The present study investigated the effect of seven different cytokines on the expression of a-smooth muscle actin (a-SMA) in cultured human tenons capsule fibroblasts. Second passage HTF were stimulated with the following cytokines at various concentrations (PDGF-AA 0.01 - 100 ng/ml, PDGF-AB 0.01 - 100 ng/ml, PDGF-BB 0.01 - 100 ng/ml, TGF- ß1 0.0005 - 500 ng/ml, EGF 0.01 - 100 ng/ml and IL-1ß 0.001 - 10 ng/ml, b-FGF 0.01 - 100 ng/ml) for six days. Cells treated with serum-free medium served as controls. Immunohistochemistry and immunoblotting were used to detect and quantitate alpha smooth muscle actin (a-SMA) expression. Expression of a-SMA was detected by immunoblotting only when cells were stimulated with TGF-ß1 (0.5, 5, 50 and 500 ng/ml). Scanning of the lumigrams showed that the amount of a-SMA increased 1.45-fold (5 ng/ml TGF-ß1), 2.8-fold (50 ng/ml TGF-ß1) and 2.55-fold (500 ng/ml TGF-ß1) when compared to 0.5 ng/ml TGF-ß1. No a-SMA was found by immunoblotting when cells were treated with TGF-ß1 (0.005 ng/ml and 0.0005 ng/ml), PDGF-AA, PDGF-AB, PDGF-BB, EGF, bFGF, IL-1ß or with serum-free medium alone. We conclude, that from all cytokines tested, only TGF-ß1 induces myofibroblast transformation of cultured HTF at high concentrations.

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