Balaiya Sankarathi, Koushan Keyvan, McLauchlan Tatiana, Chalam K V
Department of Ophthalmology, University of Florida College of Medicine , Jacksonville, Florida.
J Ocul Pharmacol Ther. 2014 Oct;30(8):625-33. doi: 10.1089/jop.2013.0225. Epub 2014 Jul 7.
To assess the cytotoxicity of varying concentrations of Brilliant Blue Green (BBG) on human retinal pigment epithelial (HRPE) cells exposed to metal halide surgical endoilluminator (SE) at varying distances of illumination.
HRPE (ARPE-19) were exposed to 2 concentrations (0.25 and 0.5 mg/mL) of BBG and illuminated with SE for 1, 5, and 15 min. Illumination (measured with light meter) was positioned at varying distances (1 and 2.5 cm) to mimic surgical distance of illumination. Cell viability (WST-1 assay) as well as structural changes in cells was quantified.
At 1 cm distance of illumination, 0.25 mg/mL BBG decreased viability of HRPE to 89.6%±4.3%, 83.9%±10.9%, and 38.9%±5.1% of controls after 1, 5, and 15 min of exposure, respectively. Similarly, 0.5 mg/mL BBG at 1 cm distance of illumination reduced the viability of HRPE to 93.7%±2.8%, 59.6%±16%, and 34.7%±3.5% of controls. At the distance of 2.5 cm, HRPE showed improved viability; cells exposed to 0.25 mg/mL BBG maintained 98.85%±3.3%, 95.31%±7.12%, and 62.07%±3.0% of viability compared with controls after 1, 5, and 15 min of exposure. Morphometric evaluation of RPE cells showed increased width (swelling) of HRPE.
BBG at its clinically used concentration (0.25 mg/mL) during vitreoretinal surgery is safe and not toxic to HRPE for up to 5 min under focal illumination (1 cm) and up to 15 min under diffuse illumination (2.5 cm) from the commonly used SE. RESULTS of our study are useful in establishing safety parameters for the use of BBG dye in vitreoretinal surgery.
评估不同浓度的亮蓝绿（BBG）对在不同照明距离下暴露于金属卤化物手术内照明器（SE）的人视网膜色素上皮（HRPE）细胞的细胞毒性。
将HRPE（ARPE - 19）暴露于2种浓度（0.25和0.5mg/mL）的BBG，并使用SE分别照射1、5和15分钟。照明（用光度计测量）放置在不同距离（1和2.5cm）以模拟手术照明距离。对细胞活力（WST - 1测定）以及细胞的结构变化进行量化。
在1cm照明距离下，0.25mg/mL的BBG在暴露1、5和15分钟后，使HRPE的活力分别降至对照组的89.6%±4.3%、83.9%±10.9%和38.9%±5.1%。同样，在1cm照明距离下，0.5mg/mL的BBG使HRPE的活力降至对照组的93.7%±2.8%、59.6%±16%和34.7%±3.5%。在2.5cm距离处，HRPE的活力有所提高；暴露于0.25mg/mL BBG的细胞在暴露1、5和15分钟后，与对照组相比，活力分别维持在98.85%±3.3%、95.31%±7.12%和62.07%±3.0%。对RPE细胞的形态计量学评估显示HRPE的宽度增加（肿胀）。
在玻璃体视网膜手术中，临床使用浓度（0.25mg/mL）的BBG在聚焦照明（1cm）下长达5分钟以及在常用SE的漫射照明（2.5cm）下长达15分钟时对HRPE是安全且无毒的。我们的研究结果有助于确定在玻璃体视网膜手术中使用BBG染料的安全参数。
