M Knorr1, P O Denk. 1. Department of General Ophthalmology, University Eye Clinic, Tübingen, Germany.
Abstract
BACKGROUND: In order to develop new strategies for the pharmacological modulation of posttraumatic and postsurgical wound healing of the corneal stroma, the effect of Trapidil, a competitive platelet-derived growth factor (PDGF) antagonist, on the proliferation of cultured bovine stromal fibroblasts (BSF) was investigated. METHODS: BSF, obtained from explant cultures, were seeded at a cell density of 100/mm2. The effect of various concentrations of Trapidil on cell viability and cell proliferation was determined using three different culture conditions: (1) serum-free medium (WM/F12), (2) serum-containing medium (WM/F12 + 10% FCS), and (3) serum-free medium + 50 ng/ml PDGF-BB. Trapidil was added in concentrations ranging from 100 micrograms/ml to 400 micrograms/ml. Cell numbers were determined 2 and 5 days after addition of Trapidil, using a computer-based cell-counting system. Cell viability was evaluated morphologically and by means of a repopulation assay. RESULTS: Addition of Trapidil (100-400 micrograms/ml) led to a significant, dose-dependent inhibition of both serum- and PDGF-BB-induced proliferation of BSF. In contrast, treatment of quiescent BSF, cultured in serum-free medium, did not result in a significant reduction of cell number. No cytotoxic effects were observed. CONCLUSION: The results of the present study demonstrate an inhibitory effect of Trapidil on the proliferation of BSF. It can be assumed that application of Trapidil might be a useful tool in the prevention of corneal complications after trauma (e.g., scarring, astigmatism and--with respect to photorefractive procedures--formation of haze and regression of the refractive effect).
BACKGROUND: In order to develop new strategies for the pharmacological modulation of posttraumatic and postsurgical wound healing of the corneal stroma, the effect of Trapidil, a competitive platelet-derived growth factor (PDGF) antagonist, on the proliferation of cultured bovine stromal fibroblasts (BSF) was investigated. METHODS: BSF, obtained from explant cultures, were seeded at a cell density of 100/mm2. The effect of various concentrations of Trapidil on cell viability and cell proliferation was determined using three different culture conditions: (1) serum-free medium (WM/F12), (2) serum-containing medium (WM/F12 + 10% FCS), and (3) serum-free medium + 50 ng/ml PDGF-BB. Trapidil was added in concentrations ranging from 100 micrograms/ml to 400 micrograms/ml. Cell numbers were determined 2 and 5 days after addition of Trapidil, using a computer-based cell-counting system. Cell viability was evaluated morphologically and by means of a repopulation assay. RESULTS: Addition of Trapidil (100-400 micrograms/ml) led to a significant, dose-dependent inhibition of both serum- and PDGF-BB-induced proliferation of BSF. In contrast, treatment of quiescent BSF, cultured in serum-free medium, did not result in a significant reduction of cell number. No cytotoxic effects were observed. CONCLUSION: The results of the present study demonstrate an inhibitory effect of Trapidil on the proliferation of BSF. It can be assumed that application of Trapidil might be a useful tool in the prevention of corneal complications after trauma (e.g., scarring, astigmatism and--with respect to photorefractive procedures--formation of haze and regression of the refractive effect).