Abstract

SO.04.13

Influence of temporary hypothermia on endothelial cell density during organ culture preservation

Schroeter J.¹, Meltendorf C.², Ohrloff C.², Rieck P.^1
1Department of Ophthalmology, Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin; ²Department of Ophthalmology, Johann Wolfgang Goethe University, Frankfurt am Main

Objective: To evaluate temporary exposure to hypothermia for its effects on endothelial cell density of porcine corneas in dextran containing organ culture medium with regards to possible negative influences of low temperatures during the transport of corneal grafts.
Methods: Two groups of central discs from pig corneas (diameter 8 mm) were organ-cultured (MEM with 6% dextran 500) for 24 hours at 32°C. Twelve corneas per group were exposed to 4°C for 12 hours and to 21° for 48 hours each. The paired corneal discs were not treated and served as controls. After further organ culture of all corneas for 48 hours at 32°C to allow regenerative processes, corneal endothelium was stained with alizarin red S and examined by light microscopy. The endothelial cell densities were determined manually on three central images.
Results: Exposure for 12 hours to 4°C as well for 48 hours to 21°C induced an endothelial cell loss of 0.3% and 1.8%, respectively. Statistical analysis showed no significant difference (p=0.680) of the endothelial cell density between corneas exposed to 4°C and the control corneas (4166±389 cells/mm² and 4177±407 cells/mm² respectively). Despite the minor cell loss, the difference of the endothelial cell density between corneas exposed to 21°C and the control corneas (4085±260cells/mm² and 4159±312 cells/mm² respectively) was statistically significant (p=0.025).
Conclusions: Exposure of organ cultured porcine corneas in dextran containing medium to 4°C for 12 hours and 21°C for 48 hours does not compromise the endothelial cell density of donor corneas in a clinically relevant manner. A storage of corneal grafts at temperatures down to 4°C for 12 hours, as it might be the case during transports from the cornea bank to the ocular surgeon, does not seem to damage the endothelial cell layer.

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