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Tanja Stachon, Jiong Wang, Timo Eppig, Achim Langenbucher, Berthold Seitz, Nora Szentmáry; Apoptosis And Viability Of Human Corneal Endothelial Cell Cultures Following Photodynamic Therapy (pdt). Invest. Ophthalmol. Vis. Sci. 2012;53(14):6014.
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With increasing resistance of microorganisms to antibiotics, alternative therapeutic options may be needed in treatment of infectious keratitis. Photodynamic therapy (PDT) may be one potential option. The purpose of this study was to evaluate apoptosis and viability of human corneal endothelial cell cultures following photodynamic therapy (PDT).
Human corneal endothelial cell line (HCEC) was cultured on Fibronectin coated cell culture dishes in DMEM/Ham’s F12 supplemented with 5% FCS. HCEC underwent illumination (670 nm) for 13 minutes following exposure to 0, 50, 100, 150, 250, 350 and 500 nM concentrations of photosensitizer chlorine e6 (Ce6) in the culture medium for 30 minutes. Twenty-four hours following PDT the percentage of apoptotic HCEC was assessed using APO-DIRECTTM Kit by flow-cytometry (FACS) and viability of HCEC was determined using alamarBlue® assay.
Twenty-four hours after PDT the percentage of apoptotic cells increased significantly (p<0.05) at concentrations higher than 150 nM of Ce6. Cell viability decreased significantly (p<0.05) at concentrations higher than 50 nM of Ce6 using PDT. Using Ce6 or illumination only the percentage of apoptotic cells and viability remained unchanged.
Photodynamic therapy triggers apoptosis and decreases viability of human corneal endothelial cells in vitro.
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