Phototoxicity of indocyanine green and Brilliant Blue G under continuous fluorescent illumination on cultured human retinal pigment epithelial cells
Phototoxicity of indocyanine green and Brilliant Blue G under continuous fluorescent illumination on cultured human retinal pigment epithelial cells
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连续荧光照射下吲哚菁绿和亮蓝G对培养的人视网膜色素上皮细胞的光毒性
DOI：
10.1167/iovs.12-10754
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Takeuchi M
中科院分区：
文献类型：
--
作者：
Takayama K;Sato T;Karasawa Y;Sato S;Ito M;Takeuchi M
目的：我们在模拟环境光的荧光灯照射下比较吲哚菁绿色（ICG）和亮蓝G（BBG）对培养的RPE细胞的光毒性。将培养的人RPE系细胞用临床使用浓度的ICG或BBG溶液染色，并在黑暗中或在荧光灯照射下在无色培养基中培养24小时。培养后，观察细胞形态和TUNEL阳性凋亡细胞。评价细胞活力和细胞死亡率。测定BBG孵育前后的吸收光谱变化。ICG染色的细胞在光照下培养改变为椭圆形形态，凋亡细胞的数量增加，而ICG染色的细胞在黑暗中培养，和BBG染色的细胞在光照和黑暗条件下培养保持平坦的形态，而不增加凋亡细胞。细胞活力下降，细胞死亡率增加，只有在细胞染色ICG，然后在光照下培养。用临床使用浓度的十分之一的ICG染色细胞在光照下培养后不诱导细胞毒性。在孵育24小时后，保留在染色细胞中的约30%的总BBG释放到培养上清液中。荧光灯照射后BBG的吸收光谱没有发生变化。用荧光灯照射导致ICG暴露的培养RPE细胞通过凋亡而死亡，但在BBG暴露的培养RPE细胞中没有。BBG可能是一种比ICG更安全的染料，因为它具有低的光诱导细胞毒性和从染色细胞中快速洗脱的特性。
Purpose.: Wecompared the phototoxicity of indocyanine green (ICG) and Brilliant Blue G (BBG) in cultured RPE cells under fluorescent lamp illumination imitating ambient light.Methods.: Cultured human RPE line cells were stained with ICG or BBG solution at concentrations of clinical use, and cultured in a colorless medium for 24 hours in the dark or under illumination from a fluorescent lamp. After culture, cell morphology and TUNEL-positive apoptotic cells were observed. Cell viability and cell death rate were evaluated. Absorption spectral changes of BBG before and after incubation were measured.Results.: ICG-stained cells cultured under illumination changed to an oval morphology with increased number of apoptotic cells, whereas ICG-stained cells cultured in the dark, and BBG-stained cells cultured under illumination and dark conditions maintained a flat morphology without increase in apoptotic cells. Cell viability decreased and cell death rate increased only in cells stained by ICG followed by culture under illumination. Staining cells with ICG at one-tenth concentration of clinical usage induced no cytotoxicity after culture under illumination. Approximately 30% of total BBG retained in the stained cells was released into the culture supernatant after incubation for 24 hours. The absorption spectrum of BBG did not change after fluorescent light irradiation.Conclusions.: Illumination with a fluorescent lamp caused cell death via apoptosis in ICG-exposed, but not in BBG-exposed cultured RPE cells. BBG may be a safer dye than ICG because of low light-induced cytotoxicity and rapid elution from stained cells.