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Abstract
Abstract
Influence of Clotting Time and Diluent on the Epitheliotrophic Capacity of Serum Eye Drops
Liu L., Harloff S., Herminghaus P., Hartwig D., Wedel T., Geerling G.
Department of Ophthalmology, Medical University of Luebeck
Purpose: Various studies on serum eye drops for the treatment of ocular surface disorders have shown variable efficacy, when using different production protocols. We previously could show that the concentration of growth factors in autologous serum and its epitheliotrophic capacity depend on the centrifugation force used. However, other production parameters, such as the clotting time and the diluent used may determine the composition of the blood product. We therefor examined the effect of serum from healthy donors prepared after variable times of clotting and with 2 different diluents in a human corneal epithelial cell culture model.
Method: Full blood samples from 10 healthy donors were allowed to clot for 20, 60 and 120 minutes, centrifuged under standardised conditions and diluted with BSS or 0.9% saline. In these samples EGF, FGF, TGF-b1, HGF, PDGF-AB, Fibronectin as well as Vitamin A and E were quantified by means of routine ELISA- or HPLC-technology. SV-40 immortalised human corneal keratinozytes were cultured in 96-well plates at 37°C, 5% CO2 with a fully defined culture medium. At 30% confluency the culture medium was substituted by one of the test preparations. Proliferation of cell cultures was quantified by means of a luminescence-based ATP-Assay in dose-response experiments. A colony disper