Yuen Darana, Gonder John, Proulx Alain, Liu Hong, Hutnik Cindy
Department of Ophthalmology, University of Western Ontario, London, Ontario, Canada.
Am J Ophthalmol. 2009 Feb;147(2):251-259.e2. doi: 10.1016/j.ajo.2008.08.031. Epub 2008 Nov 7.
To compare the in vitro toxicity of brilliant blue G (BBG), indocyanine green (ICG), Trypan blue (TB), and Evans blue (EB) in a human retinal pigment epithelial cell line (ARPE-19) and a murine retinal ganglion/Muller glial (RGC) primary cell culture.
In vitro cell biology experimental study.
The dose-dependent toxicity of the dyes was determined by exposing each dye at four different concentrations to the two cell cultures for a short exposure (three minutes) and a medium exposure (30 minutes). The time-dependent toxicity of the dyes was also determined. All four dyes, each diluted to 1/500th of stock concentration, were applied only to the ARPE-19 cells for a prolonged exposure of two, 24, 48, and 72 hours. Cell viability was measured via a mitochondrial dehydrogenase assay.
BBG was the only dye to cause toxicity in the ARPE-19 cell line at short exposure times. BBG and TB demonstrated toxicity at medium exposure times. BBG and ICG demonstrated toxicity at long exposure times and dilute concentrations. At short exposure times, none of the dyes caused toxicity in the RGC mixed primary cultures. In contrast, at medium exposure times, all dyes except ICG demonstrated toxicity that lessened with lower concentrations.
All dyes demonstrated relatively safe viability profiles in both cell lines at surgically relevant concentrations and times. Cell toxicity could be elicited at higher concentrations and longer exposure times. ICG had a favorable viability profile at almost all of the concentrations and times tested.
比较亮蓝G(BBG)、吲哚菁绿(ICG)、台盼蓝(TB)和伊文思蓝(EB)在人视网膜色素上皮细胞系(ARPE-19)和小鼠视网膜神经节/穆勒胶质(RGC)原代细胞培养物中的体外毒性。
体外细胞生物学实验研究。
通过将每种染料以四种不同浓度分别短时间(三分钟)和中等时间(30分钟)暴露于两种细胞培养物来测定染料的剂量依赖性毒性。还测定了染料的时间依赖性毒性。将所有四种染料均稀释至原液浓度的1/500,仅对ARPE-19细胞进行长达2、24、48和72小时的长时间暴露。通过线粒体脱氢酶测定法测量细胞活力。
在短时间暴露时,BBG是唯一在ARPE-19细胞系中引起毒性的染料。在中等时间暴露时,BBG和TB表现出毒性。在长时间暴露和低浓度时,BBG和ICG表现出毒性。在短时间暴露时,没有一种染料在RGC混合原代培养物中引起毒性。相反,在中等时间暴露时,除ICG外的所有染料均表现出毒性,且浓度越低毒性越小。
在手术相关的浓度和时间下,所有染料在两种细胞系中均表现出相对安全的活力特征。在较高浓度和较长暴露时间下可引发细胞毒性。在几乎所有测试的浓度和时间下,ICG都具有良好的活力特征。