PURPOSE: To improve corneal endothelial cell survival during cold preservation by the addition of a compound that enhances cell membrane repair. METHODS: Cell survival was assessed using intracellular accumulation of the fluorescent molecule calcein as a marker for endothelial cell viability in bovine corneas stored at 4 degrees C in Optisol-GS. The effect on endothelial cell survival of artificially lowering the surface tension of the cell membrane was also assayed by the addition of a surfactant, Poloxamer 188 (PLURONIC F68), which has been shown to reseal damaged cell membranes, even under conditions where metabolism is inhibited. RESULTS: Endothelial cell survival in corneas stored at 4 degrees C in Optisol-GS was increased by the addition of 1 mg/mL Poloxamer 188. The level of dead or missing corneal endothelial cells was less than 1% for corneas stored for 6, 9, or 12 days in Optisol-GS with 1 mg/mL Poloxamer 188 at 4 degrees C. In contrast, dead or missing corneal endothelial cell percentages were increased to approximately 4% after 9 days and approximately 10% after 12 days in storage in Optisol-GS without Poloxamer 188 at 4 degrees C. CONCLUSION: The surfactant Poloxamer 188 enhances endothelial cell survival in cold-stored bovine corneas.
PURPOSE: To improve corneal endothelial cell survival during cold preservation by the addition of a compound that enhances cell membrane repair. METHODS: Cell survival was assessed using intracellular accumulation of the fluorescent molecule calcein as a marker for endothelial cell viability in bovine corneas stored at 4 degrees C in Optisol-GS. The effect on endothelial cell survival of artificially lowering the surface tension of the cell membrane was also assayed by the addition of a surfactant, Poloxamer 188 (PLURONIC F68), which has been shown to reseal damaged cell membranes, even under conditions where metabolism is inhibited. RESULTS: Endothelial cell survival in corneas stored at 4 degrees C in Optisol-GS was increased by the addition of 1 mg/mL Poloxamer 188. The level of dead or missing corneal endothelial cells was less than 1% for corneas stored for 6, 9, or 12 days in Optisol-GS with 1 mg/mL Poloxamer 188 at 4 degrees C. In contrast, dead or missing corneal endothelial cell percentages were increased to approximately 4% after 9 days and approximately 10% after 12 days in storage in Optisol-GS without Poloxamer 188 at 4 degrees C. CONCLUSION: The surfactant Poloxamer 188 enhances endothelial cell survival in cold-stored bovine corneas.