Abstract
Purpose:
To determine the effect of aflibercept on barrier function in cultured highly polarized retinal pigment epithelial cells.
Methods:
Polarized RPE cells were cultured in Boyden chamber by our method<br /> (Terasaki et al, PLos One 2013). Aflibercept was added in upper chamber (400 μg/ml).<br /> After 24 hours, cytotoxicity of polarized RPE was evaluated by TUNEL stanining and morphological analysis . Barrier function of polarized RPE cells with or without aflibercept was also measured by transepithelial volt-ohmmeter. The expression of ZO-1, a tight junction-associated molecule, was investigated by immunohistochemical staining.
Results:
After the treatment with aflibercept, number of TUNEL positive cells was not different compaired to control. <br /> No apparent morphological change was observed by electron- or light microscope. Although expression of ZO-1 was strongly expressed at cell border in control, its expression was not changed in polarized RPE cells with aflibercept. In addition, the TER in RPE cells was not significantly changed by exposure to the<br /> aflibercept compared to controls.
Conclusions:
Aflibercept did not show any toxic change on polarized RPE cells in vitro.<br /> Considering its similarity to RPE cells in vivo, aflibercept might not<br /> have a significant effect on barrier function of RPE cells at least with<br /> the clinical dosage.