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104. Jahrestagung der Deutschen Ophthalmologischen Gesellschaft 2006
Abstract
Abstract
SO.14.07 Wetting agents differ in their protection of human epithelial cells against drying Tost F.1, Paulsen K.2, Giebel J.2
1Eye Clinic and 2Institute for Anatomy and Cell Biology, Ernst-Moritz-Arndt University, Greifswald Objective: Dry eye is accompanied by disorders which affect the natural function of the corneal and conjunctival epithelial barrier. The instillation of tear substitutes should compensate the deficit in wetting and protect the mucosa against drying. In order to admit the protective effect of wetting agents to a standardised evaluation, a cell culture model was established using human epithelial cell lines.
Methods: The protection against drying was tested using pharmacological substances in a cell culture test on the conjunctival epithelial cell line Chang 1-5c-4 (series 1) and corneal cell culture line 2.040 pRSV-T (series 2). On confluent cell growth the cultures were wetted for 20 min. with Artelac® EDO, Vidisic® EDO, Vidisic® Fluid EDO, Acuolens®, Viscofresh, Hyal Drops SDU, PBS, medium. Immediately afterwards we exposed the cell cultures to a constant air flow for 0,15,30,45 min. Incubation with vitality assay alamarBlue (Biosource) absorption of the oxidised form of the dye was measured using an ELISA-Reader, in order to detect the number of live epithelial cells still present.
Results: Cell survival rates in series 1 after 0,15,30,45 min. (1.02;0.81;0.35;0.32) Artelac® EDO; (0.82;0.69; 0.63;0.54) Vidisic® EDO; (0.77;0.80;0.67;0.70) Vidisic® Fluid EDO; (0.76;0.70;0.36; 0.34) Acuolens®; (0.97;0.46;0.35;0.33) Viscofresh; (0.88;0.85;0.37;0.33) Hyal Drops SDU; (0.71;0.44;0.34;0.33) PBS; series 2 (1.03;0.84;-0.21;-0.20) Artelac® EDO; (0.89;0.92;0.93;0.86) Vidisic® EDO; (0.96;0.88;0.85;0.85) Vidisic® Fluid EDO; (1.01;0.75;-0.02;-0.03) Acuolens®; (0.98;0.17;-0.22;-0.20) Viscofresh; (0.97;0.83;0.03;-0.21) Hyal Drops SDU; (0.96;0.26;-0.24;-0.21) PBS. Extended drying time periods (30, 45 min.) led to a higher loss of live cells in most preparations. Two of the active substances tested showed a significantly better protective effect for these time periods. Loss of live epithelial cells was lowest after wetting with Vidisic® Fluid EDO and Vidisic® EDO.
Conclusions: The protective effect of pharmacological substances to the highly differentiated epithelium of the human ocular surface against drying is accessible to standardised evaluation using a cell culture test procedure on human epithelial cell lines. As the vital dye has positive redox potential and itself causes no cytotoxic effect, the measuring procedure achieves a high degree of reliability.
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