Comparative Effects of Six Intraocular Vital Dyes on Retinal Pigment Epithelial Cells
Comparative Effects of Six Intraocular Vital Dyes on Retinal Pigment Epithelial Cells
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DOI：
10.1167/iovs.09-4916
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发表时间：
2010-11-01
影响因子：
4.4
通讯作者：
Corcostegui, Gonzalo
中科院分区：
文献类型：
--
作者：
Morales, Maria-Celia;Freire, Vanesa;Corcostegui, Gonzalo
目的.评价和比较吲哚菁绿色（ICG）、吲哚菁绿色（IfCG）、台盼蓝（TB）、溴酚蓝（BrB）、专利蓝（PB）和亮蓝G（BBG）对人色素上皮细胞的作用。体外培养的ARPE-19细胞暴露于这些染料，并评价急性和慢性毒性。MTT比色法测定细胞活力，相差显微镜观察细胞形态，碘化丙啶（PI）流式细胞仪测定细胞膜通透性（CMP），DiOC（6）（3）测定线粒体膜电位（Delta Psi m）。每种研究的染料在手术剂量急性暴露后均表现出毒性。光的存在经常降低细胞活力，特别是在ICG、TB、BrB和BBG的情况下孵育3小时后测量时。ICG、IfCG和BBG诱导形态学变化。CMP和Delta Psi m在暴露于手术剂量的ICG、TB、PB和四倍手术剂量的BrB后发生改变。慢性暴露于某些染料的残留量与增殖减少甚至细胞死亡有关。谨慎的做法似乎是使用每种染料的尽可能低的剂量，以尽量减少毒性作用的风险。这一预防措施在BrB的情况下可能特别重要，其使用量不应超过0.5%。此外，手术后充分冲洗玻璃体腔以完全去除痕量染料可能至关重要，特别是在ICG的情况下，以最大限度地减少慢性毒性。（Invest Ophthalmol维斯Sci.2010;51：6018-6029）DOI：10.1167/iovs.09-4916
PURPOSE. To evaluate and compare the effects of the following dyes on human pigmented epithelial cells: indocyanine green (ICG), infracyanine green (IfCG), trypan blue (TB), bromophenol blue (BrB), patent blue (PB), and Brilliant Blue G (BBG).METHODS. ARPE-19 cells cultured in vitro were exposed to these dyes, and acute and chronic toxicity were evaluated. Cell viability was measured by colorimetry (MTT assay), morphology was observed by phase-contrast microscopy, membrane permeability (CMP) was evaluated by flow cytometry with propidium iodide (PI), and mitochondrial membrane potential (Delta Psi m) was measured with 3,3'-dihexyloxacarbocyanine (DiOC(6)(3)).RESULTS. Each of the studied dyes exhibited toxicity after acute exposure at surgical doses. The presence of light often reduced cell viability, especially when measured 3 hours after incubation in the case of ICG, TB, BrB, and BBG. Morphologic changes were induced by ICG, IfCG, and BBG. Both CMP and Delta Psi m were altered after exposure to surgical doses of ICG, TB, PB, and a fourfold surgical dose of BrB. Chronic exposure to residual amounts of some dyes was associated with reduced proliferation and even cell death.CONCLUSIONS. It appears to be prudent to use the lowest possible dose of each dye, to minimize the risk of toxic effects. This precaution may be particularly important in the case of BrB, which should not be used in excess of 0.5%. In addition, abundant irrigation of the vitreous cavity after surgery to completely remove traces of dye may be of crucial importance, particularly in the case of ICG, in minimizing chronic toxicity. (Invest Ophthalmol Vis Sci.2010;51:6018-6029) DOI:10.1167/iovs.09-4916