Vitamin E succinate inhibits proliferation and migration of retinal pigment epithelial cells in vitro: therapeutic implication for proliferative vitreoretinopathy

Abstract

Background: Retinal pigment epithelial (RPE) cells play an important role in proliferative vitreoretinopathy (PVR). Vitamin E succinate is an ester form of a potent biological antioxidant, vitamin E, and has unique effects on various cells. We examined the effect of vitamin E succinate on proliferation and migration of cultured bovine RPE cells, since these are critical steps in the development of PVR.

Methods: Bovine RPE cells were cultured in minimal essential medium (MEM) containing 10% fetal calf serum (MEM-10). Cells were incubated with MEM-10 containing 25 microM vitamin E, vitamin E succinate, butylated hydroxytoluene (BHT) or d-mannitol. Cell proliferation was assessed by counting cell numbers on days 2, 4 and 6. 3H-Thymidine uptake was also examined in RPE cells incubated with various forms of vitamin E-- vitamin E, vitamin E succinate, Trolox, gamma-tocopherol, vitamin E acetate, vitamin E phosphate, vitamin E nicotinate--or antioxidants-- BHT or d-mannitol (25 microM each). RPE cell migration was studied as follows: A small area (5 x 15 mm) of confluent cultured RPE cells was denuded using a straight razor blade and incubation was continued for 20 h with MEM-10 containing vitamin E, vitamin E succinate, gamma-tocopherol or BHT. The number of cells that migrated into the denuded area from the wound edge in each microscopic field (x20) was counted and expressed as a percentage of control (MEM-10 alone).

Results: The antioxidants, vitamin E and BHT, stimulated RPE cell proliferation and 3H-thymidine incorporation compared with the control, while vitamin E succinate significantly inhibited both proliferation and 3H-thymidine uptake (IC50, 23 microM). Other forms of vitamin E or d-mannitol had no effect. Neither vitamin E nor BHT had a significant effect on RPE cell migration (108.2% and 112.6% of control, respectively), but vitamin E succinate inhibited migration (58.3%). Cell viability, assessed by the trypan blue dye exclusion test, was not impaired by a 3-day incubation with 50 microM of vitamin E succinate.

Conclusions: An ester form of a physiological antioxidant, vitamin E succinate, inhibits RPE cell proliferation and migration without causing cellular toxicity. These findings suggest its therapeutic potential for the pharmacological treatment of PVR.