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The aim of this study was to evaluate the effects of indocyanine green (ICG), which is a fluorescent dye recently used for staining during manipulation of the internal limiting membrane (ILM) in surgery, on cultured human retinal pigment epithelial (HRPE) cells. Different concentrations, up to 0.25% (weight/volume), of ICG were added to the culture medium, which contained the cultured human RPE cells. Then the cell-growth curve and cell viability were assayed with a direct cell count and MTS cell proliferation kit, respectively. Two (2) models of experimentations, a dose-response group and a short-term application group, were designed. We obtained data for IC50 values for 24-, 48-, and 72hour ICG treatments, 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 HRPE cells. In the group of 3-day sequential incubations with ICG treatment, a 0.01% concentration significantly inhibited HRPE growth. In addition, HRPE cells were damaged immediately after ICG treatment, and the damage was ICG dose-related. In the sequential 3-day incubation, compared to the control group, the surviving HRPE cells of each treated group had the same growth rate. In conclusion, ICG has a relatively toxic effect on cultured HRPE cells, even in much lower concentrations than clinical nondilution status (0.25%). Since an ICG toxic effect on HRPE cells was found in our study (in vitro), we highly recommend staining the ILM with as low a concentration of ICG as possible, and washing out the residual intravitreal ICG as thoroughly as possible after finishing the peeling of the ILM. [ABSTRACT FROM AUTHOR]

It has been well documented that RPE cells play an important role in the formation of proliferative vitreoretinopathy (PVR). We studied the effects of four most widely used anti-proliferative drugs, i.e., 5-fluorouracil (5-FU), daunomycin (DM), mitomycin C (MMC) and dexamethasone (DEX) in an in vitro model system (cultured human RPE). The cells treated with indicated concentrations of drugs for 48 h were harvested for DNA content analysis. In addition, a time course study with constant dosage of drugs was performed. The results revealed that, after incubation for 48 h, an increased percentage (31.1%) of S phase cells was noted with exposure to MMC (10 ng/ml). It was confirmed by 5-bromodeoxyuridine (BrdU) incorporation test that this increase does not result from higher transition rate from G1 to S phase. The cells of G2/M phase markedly increased from 13.8% to 29.7% with 10 ng/ml DM. We also demonstrated that 5-FU and MMC treatment led to cell accumulation at S phase and DM treatment resulted in cell accumulation at G2/M phase, These findings were compatible with their pharmacological mechanisms. Development of an in vitro model using cultured human RPE to study the effects of various anti-proliferative drugs on cell cycle can provide a rapid, safe and inexpensive method for selection of drugs used for management of PVR. [ABSTRACT FROM AUTHOR]

The effects of various antiproliferative drugs have been tested in numerous cell types in vitro, but a comparison of effects of these drugs on cultured human retinal pigment epithelium (RPE) has not been reported. We studied the effects of four most widely used antiproliferative drugs (5-FU, daunomycin, mitomycin and dexamethasone) in a new in vitro model system (cultured human RPE). Various concentrations and exposure periods were tested in four human RPE cell lines. 5-FU showed a dose-dependant growth inhibition, with an ID[sub 50] of 5.35 ± 2.38 × 10[sup -7] M after 4 days culture. A decrease of cell number occurred relatively later, the slope of the dose-response curve was less steep than that of others. Mitomycin C showed an immediate and strong growth inhibition and cytotoxic effects, with an ID[sub 50] of 3.73 ± 0.71 × 10[sup -9] M. RPE cultured with daunomycin showed an abrupt decrease of cell number from 10[sup -9] M to 10[sup -8] M, the ID value was 1.07 ± 0.23 × 10[sup -8] M. Dexamethasone showed a biphasic effect; it stimulated cell growth at 10[sup -7] to 10[sup -6] M and inhibited cell growth at 10[sup -4] M or higher, with an ID[sub 50] of 6.05 ± 1.61 × 10[sup -3] M. The advantages and disadvantages of these drugs and the prospective clinical application of these drugs for management of proliferative vitreoretinopathy (PVR) were discussed. Development of an in vitro model using cultured human RPE to study the effects of various antiproliferative drugs can provide a rapid, safe and inexpensive method for selection of drugs used for management of proliferative vitreoretinopathy. [ABSTRACT FROM AUTHOR]