Romesh I Angunawela1, John Marshall. 1. Department of Academic Ophthalmology, Rayne Institute, St Thomas' Hospital, London, United Kingdom. romeshi@hotmail.com
Abstract
PURPOSE: To evaluate the action of mannose-6-phosphate (M6P) as an inhibitor of transforming growth factor-beta1 (TGF-beta1) and its effects on human corneal fibroblasts. SETTING: Department of Academic Ophthalmology, Rayne Institute, St. Thomas' Hospital, London, United Kingdom. METHOD: Keratocytes were cultured in serum-containing medium with added TGF-beta1 and in serum-containing medium containing TGF-beta1 with M6P. Controls consisted of cells in serum-containing medium alone and cells cultured in serum-containing medium with M6P. Differentiation of fibroblasts into myofibroblasts was detected by immunohistochemistry for alpha-smooth muscle actin. Images were captured by laser confocal microscopy. RESULTS: The differentiation of cells into myofibroblasts was significantly greater after culture in medium containing TGF-beta1 than in control cultures. Myofibroblast differentiation of cells was significantly reduced when cultured in medium containing TGF-beta1 with M6P (P<.001). CONCLUSIONS: Mannose-6-phosphate significantly reduced TGF-beta1-mediated transformation of human corneal fibroblasts into myofibroblasts; it is a potential modulator of corneal wound healing and may reduce haze after refractive surgery. FINANCIAL DISCLOSURE: Neither author has a financial or proprietary interest in any material or method mentioned. Copyright 2010 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.
PURPOSE: To evaluate the action of mannose-6-phosphate (M6P) as an inhibitor of transforming growth factor-beta1 (TGF-beta1) and its effects on human corneal fibroblasts. SETTING: Department of Academic Ophthalmology, Rayne Institute, St. Thomas' Hospital, London, United Kingdom. METHOD: Keratocytes were cultured in serum-containing medium with added TGF-beta1 and in serum-containing medium containing TGF-beta1 with M6P. Controls consisted of cells in serum-containing medium alone and cells cultured in serum-containing medium with M6P. Differentiation of fibroblasts into myofibroblasts was detected by immunohistochemistry for alpha-smooth muscle actin. Images were captured by laser confocal microscopy. RESULTS: The differentiation of cells into myofibroblasts was significantly greater after culture in medium containing TGF-beta1 than in control cultures. Myofibroblast differentiation of cells was significantly reduced when cultured in medium containing TGF-beta1 with M6P (P<.001). CONCLUSIONS:Mannose-6-phosphate significantly reduced TGF-beta1-mediated transformation of human corneal fibroblasts into myofibroblasts; it is a potential modulator of corneal wound healing and may reduce haze after refractive surgery. FINANCIAL DISCLOSURE: Neither author has a financial or proprietary interest in any material or method mentioned. Copyright 2010 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.