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Mycophenolate Mofetil - a New Anti-scarring Agent in Glaucoma Surgery?
1Heinz C., 2Hudde T., 2Heise K., 3Steuhl K. P.,
1Uni-Essen (Essen)
2Universität-Gesamthochschule Essen, Zentrum für Augenheilkunde (Essen)
3Universität-Gesamthochschule Essen, Zentrum für Augenheilkunde, Abt. für Erkrankungen des vorderen Augenabschnitts (Essen)
Purpose: In penetrating glaucoma surgery antiproliferatives like mitomycin C and 5-fluorouracil are used to prevent scarring of the filtering bleb. Exaggerated scarring or severe complications might occur. We previously demonstrated, that mycophenolate mofetil (MMF), an immunosuppressive agent in organ transplantation, inhibits human Tenon fibroblasts (HTF) in vitro. MMF blocks the inosinmonophosphate dehydrogenase of the guanosine de novo synthesis. In many cell types a salvage pathway for guanosine exists. In this work we investigated if guanosine antagonises the MMF effect on HTF and evaluated HTF migration in a model.
Method: HTF (passage 3-7) have been incubated with various concentrations of MMF with or without guanosine in 10% FCS in a 96 well plate. Proliferation was assessed at days 7, 14 and 21 by cell counts. As migration model a blade was used to draw a line in a 24 well plate and the cell layer was removed with a swab on one side. Migrations of HTF over the line were documented by photography.
Results: Incubation with 1µM of MMF showed a lower proliferation rate of 71% (SD±6,4; p<0,001) compared with the control (100%) after 14 days. When incubated with 10µM of MMF the proliferation rate was reduced to 7,7% (SD±0,9; p<0,001). Addition of guanosine antagonised this antiproliferative effect completely. In the migration model, 1 µM of MMF revealed a slower migration over the line, 10 µM stopped the migration almost entirely.
Conclusions: The antiproliferative effect of MMF is mediated by guanosine depletion. In HTF no sufficient salvage pathway exists to produce guanosine. HTF depend on guanosine de novo synthesis. Proliferation rate and migration are MMF concentration dependent. MMF could be a useful antiproliferative drug in glaucoma filtering surgery.