Chalam Kakarla V, Li Wenhua, Koushan Keyvan, Grover Sandeep, Balaiya Sankarathi
Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, FL, USA.
J Clin Med Res. 2015 Jul;7(7):517-24. doi: 10.14740/jocmr2085e. Epub 2015 May 8.
The objective of the study was to determine the safety parameters of using brilliant blue green (BBG) for chromovitrectomy by assessing the cytotoxicity of BBG on cultured retinal ganglion cells (RGCs) exposed to illumination.
RGCs were exposed to two concentrations of BBG (0.25 and 0.5 mg/mL) under metal halide illumination at varying distances (1 and 2.5 cm), intensities (990 and 2,000 Fc), and durations (1, 5 and 15 minutes). Cell viability was assessed using the WST-1 and CellTiter 96(®) AQueous One solution cell proliferation assays.
Using the WST-1 assay, with high-intensity illumination, viability of RGCs ranged from 97.5±16.4% of controls with minimum BBG and light exposure (0.25 mg/mL BBG and illuminated for 1 minute at 2.5 cm distance) to 53.1±11.3% of controls with maximum BBG and light exposure (0.50 mg/mL and illuminated for 15 minutes at 1 cm distance; P < 0.01). With medium-intensity illumination, RGCs showed better viability, ranging from 95.1±7.2% of controls with minimum BBG and light exposure to 72.3±12.8% of controls with maximum BBG and light exposure. CellTiter 96(®) AQueous One assay showed similar results.
RGCs seem to safely tolerate up to 5 minutes of exposure to 0.5 mg/mL BBG under diffuse medium-intensity illumination (990 Fc).
本研究的目的是通过评估亮蓝绿(BBG)对暴露于光照下的培养视网膜神经节细胞(RGCs)的细胞毒性,来确定在脉络膜视网膜切除术使用BBG的安全参数。
在金属卤化物光照下,将RGCs暴露于两种浓度的BBG(0.25和0.5mg/mL),光照距离分别为1和2.5cm,强度分别为990和2000英尺烛光(Fc),持续时间分别为1、5和15分钟。使用WST-1和CellTiter 96® AQueous One溶液细胞增殖试验评估细胞活力。
使用WST-1试验,在高强度光照下,RGCs的活力范围为:在最低BBG和光照条件下(0.25mg/mL BBG,在2.5cm距离处光照1分钟)为对照组的97.5±16.4%,到在最高BBG和光照条件下(0.50mg/mL,在1cm距离处光照15分钟)为对照组的53.1±11.3%;P<0.01。在中等强度光照下,RGCs表现出更好的活力,范围为:在最低BBG和光照条件下为对照组的95.1±7.2%,到在最高BBG和光照条件下为对照组的72.3±12.8%。CellTiter 96® AQueous One试验显示了类似的结果。
在漫射中等强度光照(990 Fc)下,RGCs似乎能够安全耐受长达5分钟的0.5mg/mL BBG暴露。