Institute of Transfusion Medicine, University Tissue Bank, Cornea Bank Berlin, Charité - Universitätsmedizin Berlin, Campus Mitte, Charitéplatz 1, 10117, Berlin, Germany,
Graefes Arch Clin Exp Ophthalmol. 2015 May;253(5):753-8. doi: 10.1007/s00417-014-2903-0. Epub 2015 Jan 10.
To evaluate temporary exposure to hyperthermia for its impact on endothelial cell density of porcine corneas in organ culture medium containing dextran with regards to possible negative influences of high temperatures during the storage and transport of corneal grafts. Four groups of central discs (diameter 8 mm) from the corneas of both eyes in 40 pigs were first organ-cultured (MEM with 6% dextran 500) for 24 h at 32°C. Ten corneas were then exposed to 40°C in group 1, to 42°C in group 2, to 44°C in group 3, and to 50°C in group 4 for 12 h each. The paired corneal discs for all groups were not treated, stored at 32°C and served as controls. After further organ culture of all corneas for 48 h 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 on three central images using a system for the automatic estimation of morphometric parameters of corneal endothelium. Exposure for 12 h to 40°C as well as to 42°C induced no endothelial cell loss. Statistical analysis showed no significant difference of the endothelial cell density between corneas exposed to 40°C and 42°C and the control corneas (40°C treatment: 4736 ± 426 cells/mm(2) and control: 4762 ± 344 cells/mm(2), p = 0.74; 42°C treatment: 4240 ± 363 cells/mm(2) and control: 4176 ± 448 cells/mm(2), p = 0.40). Exposure to 44°C and 50°C lead to total necrosis of the endothelial cell layer. Exposure of organ cultured porcine corneas in dextran containing medium up to 42°C for 12 h does not compromise the endothelial cell density in a clinically relevant manner. Temperatures above 42°C, as it might be the case during transports from the cornea bank to the ophthalmic surgeon, must be strictly avoided as they damage the endothelial cell layer.
为了评估暂时暴露于热疗对猪眼角膜组织培养物中内皮细胞密度的影响,以便研究在角膜移植物的储存和运输过程中,高温可能带来的负面影响。将来自 40 只猪双眼的中央角膜环(直径 8mm)分为 4 组,首先在 32°C 下用含有 6%右旋糖酐 500 的 MEM 进行 24 小时的器官培养。然后,第 1 组的 10 只眼角膜暴露于 40°C,第 2 组暴露于 42°C,第 3 组暴露于 44°C,第 4 组暴露于 50°C,持续 12 小时。所有组的配对角膜盘均未进行处理,储存在 32°C 下作为对照。所有角膜在 32°C 下进一步器官培养 48 小时,以允许再生过程,然后用茜素红 S 对角膜内皮进行染色,并在光镜下进行检查。使用角膜内皮形态计量学参数自动评估系统,在三个中央图像上测定内皮细胞密度。暴露于 40°C 和 42°C 12 小时不会引起内皮细胞丢失。统计分析显示,暴露于 40°C 和 42°C 的角膜与对照角膜的内皮细胞密度无显著差异(40°C 处理:4736±426 个细胞/mm²和对照:4762±344 个细胞/mm²,p=0.74;42°C 处理:4240±363 个细胞/mm²和对照:4176±448 个细胞/mm²,p=0.40)。暴露于 44°C 和 50°C 导致内皮细胞层完全坏死。将猪眼角膜在含有右旋糖酐的培养基中培养至 42°C 并暴露 12 小时不会以临床相关的方式损害内皮细胞密度。必须严格避免高于 42°C 的温度,因为在从角膜银行到眼科医生的运输过程中可能会出现这种情况,因为它们会损害内皮细胞层。