gms | German Medical Science

To investigate the effect of alkylphosphocholines (APCs) on human Tenon fibroblast (HTF) migration, cellular toxicity and the mechanism of action involved. HTF cells are known to play a crucial role in scarring of the filtering bleb site after glaucoma filtration surgery.

HTFs were isolated from tissue samples of five consented patients obtained during surgery and cultured in DMEM, 10% FCS. After splitting, HTFs were treated with one APC (C18:1-PC, C20:1-PC, C21:1-PC or C22:1-PC, respectively) in different concentrations spanning the 50% inhibitory concentration (IC₅₀) as determined previously. Cell migration was assessed by a modification of the Boyden's chamber method in microchemotaxis chambers with fibronectin coated polycarbonate filters. The lower half of the chamber was filled with DMEM/TGFbeta2. HTFs mixed with APCs in four concentrations (0.1, 1.0, 10 or 30 μM) were added above. Toxicity was assessed by the trypan blue exclusion test. For analysis of the mechanism of action, protein kinase C (PKC) activity was measured by a radioactive assay based on the measurement of ³²P-labeled phosphate transfer to a PKC specific peptide that can be captured on phosphocellulose filters.

All four APCs are able to inhibit HTF migration in vitro in a dose-dependent manner. Trypan blue staining revealed a toxicity within control limits in the concentration interval tested. PKC activity was significantly reduced by all four APCs within the concentration interval tested.

APCs inhibit HTF migration in vitro at non-toxic concentrations. Their mechanism of action seems to involve the inhibition of the PKC pathway. APCs might be a useful candidate as antiscarring agents in glaucoma filtration surgery.