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R. Migon, L. C. Zacharias, M. F. Estrago-Franco, G. M. Seigel, M. C. Kenney, B. D. Kuppermann; Effect of Silicone Oil on Retinal Cells in Culture. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5000.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the effect of 1000 and 5000 centistoke (cs) silicone oil (SO) in human retinal pigment epithelial (ARPE- 19) and rat retinal neurosensory (R28) cells in vitro.
ARPE-19 and R28 cells were plated into the superior compartment of 6-well 0.4um pore transwell inserts (Corning Inc, Corning, NY). Cells were grown to 80% confluency then transferred to serum-free media for 24 hours to avoid cell proliferation. SO 1000cs (Alcon, Fort Worth, Texas) or 5000cs (Bausch & Lomb, Rochester, NY) was added directly onto the cells or after a 100ul protective layer of culture media had been placed onto the cells to simulate the physiologic conditions of the superior and inferior retina, respectively. The inferior compartment of the insert was filled with culture media. Cell viability was analyzed after 24, 48, or 120 hours of SO exposure for ARPE-19 cells and after 24 or 48 hours for R28 cells. Cell viability was assessed by manual count using a Hemacytometer (Reichert Bright Line, Buffalo, NY), and results were normalized to untreated controls set to reference level of 100%.
ARPE-19 cells exposed to 1000cs SO showed significantly reduced cell viability after 48 and 120 hours when SO was added directly onto cells: 64.43 ± 9.50% and 72.81 ± 0.96% at 48 and 120 hours vs 100% for untreated cells, p<0.05. Cell viability was also reduced for cells treated with 5000cs SO directly on cells after 24 and 120 hours: 54.42 ± 12.76% and 76.91 ± 9.27% vs untreated controls, p<0.05. Cell viability was similarly reduced in ARPE-19 cells treated with SO placed over a 100ul protective layer of culture media (though not statistically significant due to larger sd): 59.73 ± 23.24% (p=0.14) and 77.42 ± 16.45% (p=0.10) for 1000cs and 5000cs at 120 hours. R28 cells treated with 1000cs or 5000cs SO showed a significant reduction in cells treated with either a protective aqueous layer then SO or with SO directly onto the cells: 52.33 ± 7.69% and 60.95 ± 6.33% for protective layer or direct 1000cs SO, respectively; 56.93 ± 16.48% and 56.43 ± 12.09% for protective layer or direct 5000cs SO, p<0.05; untreated controls 100% after 24 hours) or after 48 hours: 63.55 ± 21.99% and 39.91 ± 20.20% for protective layer or direct 1000cs SO; 56.60 ± 19.31% and 44.48 ± 9.84% for protective layer or direct 5000cs SO, p<0.05 compared to untreated controls.
Silicone oil 1000cs and 5000cs appear to reduce the number of viable retinal cells in vitro. A layer of fluid simulating the aqueous miniscus conditions observed in the inferior retina of a silicone-oil filled eye did not appear to be protective in this cell culture model.
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