Majmudar Parag A, Johnson Lauren
*Department of Ophthalmology, Rush University Medical Center, Chicago, IL; and †Eversight Eye Bank, Ann Arbor, MI.
Cornea. 2017 Feb;36(2):217-221. doi: 10.1097/ICO.0000000000001074.
The purpose of this study was to determine the correct concentration and duration of application of trypan blue dye to Descemet membrane endothelial keratoplasty (DMEK) donor corneal tissue before transplant to enhance visibility while minimizing damage to the tissue.
Corneas were precut for DMEK using an 8-mm punch, and each was stained with either 0.06% or 0.15% trypan blue for a duration of 1, 3, or 5 minutes. After staining, each tissue was placed in balanced salt solution and observed to determine how quickly the stain faded from the tissue using a grading system. Each cornea then underwent slit-lamp evaluation, specular microscopy, and assessment of the level of stain retained.
Tissue processed with 0.06% trypan blue for 1 minute regressed to a level 1 stain at an average of 109 minutes; tissue stained for 3 and 5 minutes regressed to a level 2 to 3 stain by 110 minutes. Endothelial cell density (ECD) was not affected when compared with prestaining measurements. Tissue processed with 0.15% trypan blue for 1 and 3 minutes regressed only to a level 2 to 3 stain at an average of 130 minutes; there was no change in ECD when stained for 1 to 3 minutes. Tissue stained for 5 minutes with 0.15% trypan blue regressed to a level 3 to 4 stain by 130 minutes. However, ECD was decreased by 11% when compared with prestaining measurements (P = 0.058).
Staining of DMEK donor tissue with VisionBlue (0.06%) up to 5 minutes is effective (visibility of staining is maintained for a sufficient period) and safe (no adverse effect on ECD). The higher concentration of trypan blue (0.15%, MembraneBlue) may be suggested during the learning curve of the DMEK surgeon, as it allowed a higher level of stain without adverse effects but only if used for 1 to 3 minutes; a longer staining time of 5 minutes resulted in a decrease in ECD, which approached statistical significance. Further study is required on use of the higher concentration of VisionBlue. The recent use of the "S" stamp may further help surgeons orient the graft correctly, thus requiring less intense trypan blue staining.
本研究的目的是确定在移植前将台盼蓝染料应用于Descemet膜内皮角膜移植术（DMEK）供体角膜组织的正确浓度和持续时间，以提高可见度，同时尽量减少对组织的损伤。
使用8毫米穿孔器为DMEK预先切割角膜，每个角膜分别用0.06%或0.15%的台盼蓝染色1、3或5分钟。染色后，将每个组织置于平衡盐溶液中，并使用分级系统观察以确定染料从组织中褪色的速度。然后对每个角膜进行裂隙灯评估、镜面显微镜检查以及对保留的染色水平进行评估。
用0.06%台盼蓝染色1分钟的组织平均在109分钟时退回到1级染色；染色3分钟和5分钟的组织在110分钟时退回到2至3级染色。与染色前测量相比，内皮细胞密度（ECD）未受影响。用0.15%台盼蓝染色1分钟和3分钟的组织平均在130分钟时仅退回到2至3级染色；染色1至3分钟时ECD没有变化。用0.1�%台盼蓝染色5分钟的组织在130分钟时退回到3至4级染色。然而，与染色前测量相比，ECD降低了11%（P = 0.058）。
用VisionBlue（0.06%）对DMEK供体组织染色长达5分钟是有效的（染色可见度保持足够长的时间）且安全的（对ECD无不良影响）。在DMEK手术医生的学习曲线期间，可能建议使用较高浓度的台盼蓝（0.15%，MembraneBlue），因为它能产生较高水平的染色且无不良影响，但前提是仅使用1至3分钟；5分钟的较长染色时间导致ECD降低，接近统计学意义。需要对使用较高浓度的VisionBlue进行进一步研究。最近使用“S”标记可能会进一步帮助手术医生正确定位移植物，从而减少对台盼蓝染色强度的需求。
