Weber Isabell P, Rana Mrinal, Thomas Peter B M, Dimov Ivan B, Franze Kristian, Rajan Madhavan S
Department of Physiology, Development and Neuroscience, Anatomy Building, University of Cambridge, Cambridge, United Kingdom.
Department of Ophthalmology, Addenbrookes Hospital, Cambridge University Hospitals NHS Trust, Cambridge, United Kingdom.
PLoS One. 2017 Sep 13;12(9):e0184375. doi: 10.1371/journal.pone.0184375. eCollection 2017.
The purpose of this study was to evaluate the effects of vital dyes on human Descemet's membranes (DMs) and endothelia. DMs of 25 human cadaveric corneas with research consent were treated with dyes routinely used in Descemet membrane endothelial keratoplasty (DMEK), 0.05% Trypan blue (TB) or a combination of 0.15% Trypan blue, 0.025% Brilliant blue and 4% Polyethylene glycol (commercial name Membrane Blue Dual; MB). The effects of these two dyes on (i) endothelial cell viability, (ii) DM mechanical properties as assessed by atomic force microscopy, and iii) qualitative DM dye retention were tested for two varying exposure times (one or four minutes). No significant differences in cell toxicity were observed between treatments with TB and MB at the two different exposure times (P = 0.21). Further, both dyes led to a significant increase in DM stiffness: exposure to TB and MB for one minute increased the apparent elastic modulus of the DM by 11.2% (P = 810-3) and 17.7%, respectively (P = 410-6). A four-minute exposure led to an increase of 8.6% for TB (P = 0.004) and 13.6% for MB (P = 0.03). Finally, at 25 minutes, the dye retention of the DM was considerably better for MB compared to TB. Taken together, a one-minute exposure to MB was found to improve DM visibility compared to TB, with a significant increase in DM stiffness and without detrimental effects on endothelial cell viability. The use of MB could therefore improve (i) visibility of the DM scroll, and (ii) intraoperative unfolding, enhancing the probability of successful DMEK surgery.
本研究的目的是评估活性染料对人Descemet膜（DM）和内皮细胞的影响。经研究同意，对25只人尸体角膜的DM用Descemet膜内皮角膜移植术（DMEK）中常用的染料进行处理，即0.05%台盼蓝（TB）或0.15%台盼蓝、0.025%亮蓝和4%聚乙二醇的组合（商品名Membrane Blue Dual；MB）。测试了这两种染料在两个不同暴露时间（1分钟或4分钟）对（i）内皮细胞活力、（ii）通过原子力显微镜评估的DM力学性能以及（iii）DM染料保留质量的影响。在两个不同暴露时间，TB和MB处理之间未观察到细胞毒性的显著差异（P = 0.21）。此外，两种染料均导致DM硬度显著增加：暴露于TB和MB 1分钟，DM的表观弹性模量分别增加11.2%（P = 8×10⁻³）和17.7%（P = 4×10⁻⁶）。暴露4分钟，TB增加8.6%（P = 0.004），MB增加13.6%（P = 0.03）。最后，在25分钟时，与TB相比，MB在DM上的染料保留明显更好。综上所述，发现与TB相比，暴露于MB 1分钟可提高DM的可见性，显著增加DM硬度且对内皮细胞活力无有害影响。因此，使用MB可改善（i）DM卷的可见性，以及（ii）术中展开，提高DMEK手术成功的概率。
