Wen-Chuan Wu1 , and Dan-Ning Hu 2
    高雄医科大学
    1. Department of Ophthalmology, Kaohsiung Medical University, Kaohsiung, Taiwan
    2. Tissue Culture Center, New York Eye and Eat Infirmary, New York Medical  College, New York, NY

    Purpose  Previous work has shown that indocyanine green (ICG) – assisted peeling of the internal limiting membrane during macular hole surgery may damage the retinal pigment epithelium (RPE). The aim of this study was to evaluate the direct toxic effects and phototoxic effects of ICG in cultured human RPE.

    Methods  In the first experiment, different levels concentration, up to 0.25% (weight / volume) of ICG were added to the culture medium. The cell-growth curve and cell viability were assayed with direct cell count and MTS cell proliferation kit, respectively. In the second experiment, RPE cells were exposed to ICG (0.5%, 5min) followed by light irradiation at different doses of light energy (1.0, 3.0, and 10.0 j/cm2). After 48 hours, cells were collected and stained with trypan blue to obtain the number of viable and nonviable cells in different groups.

    Results  In the first experiment, we obtained data for IC50 values for 24-, 48-, and 72- hours of ICG treatment, which were 0.062%, 0.041%, and 0.035%, respectively. This implies that the longer the culture duration, the lower the concentration of ICG, which can inhibit the human RPE cells. In the second experiment, cultures exposed to ICG without light irradiation showed a significant decrease of viable cells (-13.3%) and an increase of nonviable cells (x 2.5 fold) compared with cultures not exposed to either ICG or light, indicating the presence of direct toxic effects of ICG. In cultures exposed to ICG plus light irradiation (10.0 j/cm2), viable cells decreased significantly (-45.0 %) and nonviable cells increased significantly (x 4.4 fold) compared with cultures exposed to ICG alone. The damage to the RPE cells depended on the does of light (1.0 – 10.0 j/cm2), indicating that ICG has a phototoxic effect as well as a toxic one.

    Conclusion  ICG has both direct toxic and phototoxic effects on human RPE cells in vitro.

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