Department of Ophthalmology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, Zhejiang Province, People's Republic of China.
Department of Ophthalmology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, Zhejiang Province, People's Republic of China.
Biomed Pharmacother. 2020 Dec;132:110790. doi: 10.1016/j.biopha.2020.110790. Epub 2020 Oct 6.
This study investigated the in vitro effect of various vital dyes in common clinical use on human Müller cell viability, and it compared the toxicity of these dyes using a cell culture model. Müller cells were exposed to a series of concentrations (1 %, 0.5 %, 0.25 %, and 0.125 % or 12.9 mM, 6.45 mM, 3.22 mM and 1.61 mM) of Indocyanine green (ICG) for 2, 24, 48, and 72 h. Similarly, groups of Müller cells were stained with "Heavy" brilliant blue G (HBBG), Trypan blue (TB) (0.15 %, or 1.56 mM), Membrane-blue-dual (MBD), and ICG (0.25 %, or 3.22 mM) or BBG (0.025 %, or 0.3 mM) with glucose (GS) (50 %, 66 % and 75 % or 2.78 M, 3.67 M and 4.17 M) for 30, 60, and 120 s. Cell viability was measured with the Cell Counting Kit-8 (CCK-8) and Lactate Dehydrogenase (LDH) release assays. We found that high stain concentration and long exposure time resulted in increased toxicity to Müller cells. Nevertheless, ICG seemed to be safe at the clinically relevant concentration of 0.25 % (3.22 mM) in the short time of exposure. TB was safer than both HBBG and MBD, especially HBBG. Hypertonic GS as a dilution was not safe for Müller cells, and the negative effect was more obvious in 0.025 % (0.3 mM) BBG than that in 0.25 % (3.22 mM) ICG. This is the first report to observe the cytotoxicity of commonly used stains in clinical on human Müller cells in vitro, and to provide some basis for further studies, including in vivo investigation.
本研究旨在探讨临床上常用的各种活细胞染料在体外对人 Müller 细胞活力的影响,并采用细胞培养模型比较这些染料的毒性。将 Müller 细胞暴露于一系列浓度(1%、0.5%、0.25%和 0.125%或 12.9mM、6.45mM、3.22mM 和 1.61mM)的吲哚菁绿(ICG)中 2、24、48 和 72 小时。同样,将一组 Müller 细胞用“重”亮蓝 G(HBBG)、台盼蓝(TB)(0.15%或 1.56mM)、Membrane-blue-dual(MBD)和 ICG(0.25%或 3.22mM)或 BBG(0.025%或 0.3mM)染色,并用葡萄糖(GS)(50%、66%和 75%或 2.78M、3.67M 和 4.17M)孵育 30、60 和 120 秒。通过细胞计数试剂盒-8(CCK-8)和乳酸脱氢酶(LDH)释放测定法测量细胞活力。我们发现高染色浓度和长时间暴露会导致 Müller 细胞毒性增加。然而,在短时间暴露于临床相关浓度 0.25%(3.22mM)的情况下,ICG 似乎是安全的。TB 比 HBBG 和 MBD 都更安全,尤其是 HBBG。高渗 GS 作为稀释剂对 Müller 细胞不安全,0.025%(0.3mM)BBG 的负作用比 0.25%(3.22mM)ICG 更明显。这是首次观察到临床上常用染色剂在体外对人 Müller 细胞的细胞毒性,并为进一步研究(包括体内研究)提供了一些依据。