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Eric Miller, Anne Gemensky-Metzler, David Wilkie, Carmen Colitz, Heather Chandler; Effects of Various Antioxidants on Lens Epithelial Cells In Vitro and Ex Vivo. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2979. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To determine if three antioxidants, grape seed extract (GSE), lutein, and omega-3 fatty acids (O3FA), alter oxidative stress, migration, and proliferation in lens epithelial cells (LEC).
An antioxidant reductive capacity assay determined the antioxidant capability of the antioxidants. A DCF assay evaluated the ability of these antioxidants to reduce reactive oxygen species (ROS) production in UV stressed and unstressed LEC’s. Arrays were used to determine changes in protein expression following treatment with the antioxidants. LEC migration and proliferation were evaluated utilizing an ex vivo model of posterior capsular opacification (PCO).
The antioxidant reductive capacity of GSE surpassed that of the positive control, while lutein and O3FA showed little reducing capacity. The DCF assay corroborated this data; GSE reduced ROS production in both UV stressed and unstressed LEC cell cultures compared to the positive control, lutein appeared to be pro-oxidative and O3FA had little to no ability to reduce ROS. Protein arrays showed GSE decreased expression of IL-6, IL-8, CCL3, and CCL5 compared to controls. Lutein and O3FA showed an increase or no change in expression of the various proteins as compared to controls. The ex vivo PCO model demonstrated an increase in LEC number and migration resulting in an increased area of the posterior lens capsule covered following treatment with O3FA, while GSE and lutein treatments were similar to controls.
Only GSE showed substantial antioxidant capabilities with the ability to reduce ROS generation. Lutein and O3FA demonstrated no antioxidant abilities and lutein proved to be pro-oxidative in vitro. Following treatment with antioxidants, unstressed LEC’s showed altered expression of cytokines known to influence redox signaling, migration, and proliferation. O3FA increased cell proliferation and migration in the ex vivo PCO model while GSE and lutein demonstrated little effect. Careful consideration should be given to any conclusion about the effects of these antioxidants on LEC’s due to findings of variable and limited reducing power.
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