Department of Ophthalmology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
Clin Exp Ophthalmol. 2013 Jul;41(5):484-90. doi: 10.1111/ceo.12018. Epub 2012 Dec 10.
To determine the time-dependent toxicity of Trypan blue at 0.06% concentration in cultured human trabecular meshwork cells.
Human trabecular meshwork cells cultured in vitro were exposed to Trypan blue and acute toxicity was evaluated. Cells were exposed for 5, 30, 60 s and for 5, 15, 30, 60 min to a commercially available Trypan blue preparation (Vision Blue; DORC International Zuidland, The Netherlands). Morphology was observed by phase-contrast microscopy and cell viability was measured using Hoechst 33342 (Intergen; Purchase, NY, USA) and propidium iodide assays to determine the percentage of living and dead cells.
Morphological changes occurred mainly after 5 min of exposure to Trypan blue. Viability was 96.0% ± 3.6%, 94.8% ± 3.5% and 92.5% ± 4.4% after 5, 30 and 60 s of exposure, respectively; a significant toxic effect of Trypan blue was observed after 5, 15, 30 and 60 min of exposure with a viability of 85.0% ± 3.7%, 77.0% ± 6.7%, 70.8% ± 5.9% and 68.3% ± 8.7%, respectively.
Trypan blue is not toxic, in terms of cell viability, over an exposure time of up to 60 s; however, further exposure results in a gradual increase in damage of cultured human trabecular meshwork cells.
确定 0.06%浓度锥虫蓝在体外培养的人眼小梁细胞中的时间依赖性毒性。
将体外培养的人眼小梁细胞暴露于锥虫蓝中，并评估急性毒性。将细胞暴露于市售锥虫蓝制剂（Vision Blue；DORC International Zuidland，荷兰）5、30、60 秒和 5、15、30、60 分钟。通过相差显微镜观察形态，并用 Hoechst 33342（Intergen；Purchase，NY，USA）和碘化丙啶测定法测定活细胞和死细胞的百分比来测量细胞活力。
形态变化主要发生在暴露于锥虫蓝 5 分钟后。暴露于锥虫蓝 5、30 和 60 秒后，细胞活力分别为 96.0%±3.6%、94.8%±3.5%和 92.5%±4.4%；暴露于锥虫蓝 5、15、30 和 60 分钟后，细胞活力显著降低，分别为 85.0%±3.7%、77.0%±6.7%、70.8%±5.9%和 68.3%±8.7%。
在暴露时间不超过 60 秒的情况下，锥虫蓝在细胞活力方面没有毒性；然而，进一步的暴露会导致培养的人眼小梁细胞逐渐受损。
