A murine cell culture model for post-trabeculectomy anfibrotic treatment: Induction of apoptosis by Cyclosporin.

Abstract

PURPOSE Experimental trials aimed at the research of selective antifibrotic agents are under development for the alternative treatment of glaucoma patients who are usually considered high-risk post-surgical individuals after trabeculectomy. Authors present here an in vitro model system for the treatment of post-trabeculectomy patients. The study is aimed at the evaluation of different drugs in a mouse fibroblast model. METHODS The antifibrotic activity of Cyclosporin A, Interferon 2alpha, 5-Fluorouracyl was investigated on 3T6 cells in culture. Cell viability and proliferation was assessed after drug treatment. Molecular analysis of DNA degradation was evaluated by means of radioactive labeling and gel electrophoresis. RESULTS The three drugs were shown to affect cell proliferation and viability in a differential fashion. However, only Cyclosporin A was able to control cell proliferation, inducing apoptosis. This phenomenon was reduced by supplementation of trolox, a compound known to inhibit programmed cell death. These results strongly suggest that this model system might be useful as a test of pharmacological functionality. CONCLUSION A rapid and efficient model system is described for the assessment of cell viability and proliferation after treatment with agents of potential pharmacological use. Cyclosporin A induces a significant apoptosis. This is important for the negative control of fibrotic degeneration in post-trabeculectomy that is required for successful surgery in glaucoma patients. Therefore, Cyclosporin A might become a clinically interesting drug for the antifibrotic treatment of post-trabeculectomy.