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An evaluation of novel vital dyes for intraocular surgery.

机译:对眼内手术的新型重要染料的评估。

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PURPOSE: To evaluate systematically the staining characteristics and safety of potential new dyes for intraocular surgery. METHODS: Six dyes were included in the investigation: light green SF (LGSF) yellowish, E68, bromophenol blue (BPB), Chicago blue (CB), rhodamine 6G, rhodulinblau-basic 3 (RDB-B3). All dyes were dissolved and diluted in a balanced saline saline solution. The light-absorbing properties of each dye were measured at a concentration of 0.05% between 200 and 1000 nm. Staining characteristics were examined by staining lens capsule tissue and epiretinal membranes (ERMs), removed intraoperatively, with dye concentrations of 1.0%, 0.5%, 0.2%, and 0.05%. Enucleated porcine eyes (postmortem time, 9 hours) were also stained. Dye-related toxicity was evaluated by a colorimetric test (MTT) measuring the inhibition of retinal pigment epithelium (RPE) cell proliferation (ARPE-19 and primary human RPE cells, passages 3-6). Cell viability was also quantified based on a two-color fluorescence cell-viability assay. Dyes were investigated in concentrations of 0.2% and 0.02%. RESULTS: All dyes investigated in this study stained human lens capsules, removed intraoperatively; ERMs, peeled during macular pucker surgery; and enucleated porcine eyes, depending on the concentration applied. The long-wavelength absorption maximum of the dyes was within the range of 527 to 655 nm at concentrations of 0.05%. Rhodamine G6 and RDB-B3 showed adverse effects on ARPE-19 cell proliferation at a concentration of 0.2% and were excluded from further investigation in primary RPE cells. The remaining four dyes showed no toxic effect on ARPE-19 and primary RPE cell proliferation at concentrations of 0.2% and 0.02%. Cell viability was affected by LGSF yellowish (0.2%) and CB (0.2% and 0.02%). Two dyes (E68 and BPB) showed no relevant toxicity in vitro. CONCLUSIONS: The systematic evaluation of dyes for intraocular use seems mandatory. In this study four dyes were identified with effective staining characteristics, with two of these dyes having no detectable toxic effect on RPE cells in vitro.
机译:目的:系统评价潜在的新型染料用于眼内手术的染色特性和安全性。方法:研究中包括六种染料:浅绿色SF(LGSF)淡黄色,E68,溴酚蓝(BPB),芝加哥蓝(CB),若丹明6G,若丹明碱性3(RDB-B3)。将所有染料溶解并在平衡盐水溶液中稀释。在200和1000 nm之间的浓度为0.05%的情况下测量每种染料的吸光性能。通过染色术中去除的晶状体囊组织和视网膜前膜(ERM)来检查染色特性,染料浓度为1.0%,0.5%,0.2%和0.05%。去核猪眼(死后9小时)也被染色。通过比色测试(MTT)评估了与染料相关的毒性,该比色测试测量了视网膜色素上皮(RPE)细胞增殖的抑制作用(ARPE-19和原代人RPE细胞,第3-6代)。还基于双色荧光细胞活力测定法定量了细胞活力。研究了染料的浓度为0.2%和0.02%。结果:本研究中研究的所有染料均将人晶状体囊染色,并在术中取出。 ERM,在黄斑皱褶手术中剥落;和去核猪眼,取决于所用浓度。在0.05%的浓度下,染料的长波吸收最大值在527至655nm的范围内。若丹明G6和RDB-B3浓度为0.2%时,对ARPE-19细胞增殖具有不利影响,因此未在原代RPE细胞中进行进一步研究。其余四种染料在0.2%和0.02%的浓度下对ARPE-19和原代RPE细胞的增殖没有毒性作用。细胞活力受LGSF微黄(0.2%)和CB(0.2%和0.02%)影响。两种染料(E68和BPB)在体外没有相关毒性。结论:眼内使用染料的系统评价似乎是强制性的。在这项研究中,鉴定了四种具有有效染色特性的染料,其中两种染料在体外对RPE细胞没有可检测的毒性作用。

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