PURPOSE: To examine the in vitro effects of bile acids and salts on the viability, growth and morphology of human Tenon's capsule fibroblasts. METHODS: Human Tenon's capsule fibroblasts were grown from explants in culture and used between passages 3 and 5. Fibroblasts (n = 3) were treated in triplicate with deoxycholic acid (DA), sodium deoxycholate (NaD) and cholic acid (CA) at concentrations between 50 microM and 1 mM. Cell viability and growth were assessed with trypan blue staining and haemocytometer counting, and a colorimetric (MTT) assay. Morphology was assessed with light microscopy and cresyl violet staining. RESULTS: A dose-dependent decrease in viability was observed following bile acid treatment (24 and 48 h) where the effects of DA > NaD > CA. The LD50 values at 48 h for DA, NaD and CA were 300 microM, 400 microM and 720 microM respectively. Cell growth following bile acid treatment was reduced compared with controls. Fibroblasts treated with bile acids displayed a loss of normal spindle-shaped morphology and multiple processes, some with varicosities, extended from many cells. These changes were dose-dependent, and at higher concentrations cells became detached and were non-viable (trypan-blue-positive). Extensive cell death was observed in cultures treated with DA 400 microM, NaD 500 microM and CA 800 microM. CONCLUSIONS: Lower doses of bile acids and salts reduced in vitro fibroblast growth associated with morphological changes; higher doses induced fibroblast cytotoxicity. These observations suggest that bile acids and salts, in particular DA, may be useful in regulating wound fibrosis following trabeculectomy surgery.
PURPOSE: To examine the in vitro effects of bile acids and salts on the viability, growth and morphology of human Tenon's capsule fibroblasts. METHODS:Human Tenon's capsule fibroblasts were grown from explants in culture and used between passages 3 and 5. Fibroblasts (n = 3) were treated in triplicate with deoxycholic acid (DA), sodium deoxycholate (NaD) and cholic acid (CA) at concentrations between 50 microM and 1 mM. Cell viability and growth were assessed with trypan blue staining and haemocytometer counting, and a colorimetric (MTT) assay. Morphology was assessed with light microscopy and cresyl violet staining. RESULTS: A dose-dependent decrease in viability was observed following bile acid treatment (24 and 48 h) where the effects of DA > NaD > CA. The LD50 values at 48 h for DA, NaD and CA were 300 microM, 400 microM and 720 microM respectively. Cell growth following bile acid treatment was reduced compared with controls. Fibroblasts treated with bile acids displayed a loss of normal spindle-shaped morphology and multiple processes, some with varicosities, extended from many cells. These changes were dose-dependent, and at higher concentrations cells became detached and were non-viable (trypan-blue-positive). Extensive cell death was observed in cultures treated with DA 400 microM, NaD 500 microM and CA 800 microM. CONCLUSIONS: Lower doses of bile acids and salts reduced in vitro fibroblast growth associated with morphological changes; higher doses induced fibroblast cytotoxicity. These observations suggest that bile acids and salts, in particular DA, may be useful in regulating wound fibrosis following trabeculectomy surgery.