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K. Sternberg, T. Terwee, O. Stachs, R. Guthoff, M. Löbler, K.-P. Schmitz; An ex vivo Model for Drug-Induced Secondary Cataract Prevention - Experimental Results With Disulfiram, Methotrexate and Actinomycin D. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5673.
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A clinical approach to prevent secondary cataract by inhibition or removal of lens epithelial cells involves the intraocular application of pharmacological agents. The goal of our study was to develop an ex vivo model that can be utilized to test for the effectiveness of pharmacological agents for the ablation of lens epithelial cells from the basal membrane.
Cultured human capsular rhexis specimens from standard cataract surgery were incubated for 5 minutes with Disulfiram, Methotrexate and Actinomycin D dissolved in pure water or embedded in hyaluronic acid. After drug treatment the residual viable and dead cells were differentiated by use of the Live-dead assay. Cell quantification was facilitated by staining with Hoechst-dye.
An ex vivo model was established which allows for the differentiation of drug action on lens epithelial cell ablation from the basal membrane. The Live-dead assay on untreated specimens has shown 1361+/-482 viable cells/mm2. The drug treatment reduced the number of viable cells on the specimens drastically, because it ranges between 0.44+/-0.53 % (6.0+/-7.3 cells/mm2) for Disulfiram, 0.27+/-0.50 % (3.7+/-6.9 cells/mm2) for Methotrexate and 0.07+/-0.19 % (0.1+/-0.27 cells/mm2) for Actinomycin D. Actinomycin D was slightly more potent in cell ablation than Disulfiram and Methotrexate.
The screening of drugs in the described ex vivo model can help to reduce the number of preclinical studies for secondary cataract prevention. Furthermore, a safe drug application can be achieved by using hyaluronic acid as a hydrophilic polymeric carrier.
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