The Effects of 5-Fluorouracil and Agaricus Bisporus Lectin on Regulating Retinal Pigment Epithelial Cell Wound Healing Activities

Abstract

Purpose: : Following successful surgical repair of retinal detachment, proliferative vitreoretinopathy (PVR), a modified wound healing process, may result in re-detachment of the retina. Dedifferentiated and anomalous retinal pigment epithelial (RPE) cells have been implicated to play a major role in PVR. Previous attempts using 5-fluorouracil (5-FU) as an adjunctive surgical therapy have been shown to reduce the onset of PVR but have no effect on established PVR in vivo. Agaricus bisporus lectin (ABL), a protein extracted from edible mushroom, is shown to bind dedifferentiated RPE cells. This study examines the effects of 5-FU and ABL on RPE cell wound healing processes.

Methods: : ARPE-19 cells were treated with 5-FU (0.25-25 mg/ml) for 10 min or incubated with ABL (20-90 µg/ml). Cellular proliferation rate was measured by the CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay (MTS). Adhesion assay was conducted to evaluate the effects of 5-FU on cell adhesion towards collagen type I and fibronectin. Migration assay was performed with either 5-FU or ABL. The cytotoxicity of 5-FU was determined by the live/dead assay.

Results: : Both ABL and 5-FU were found to inhibit cellular proliferation, adhesion and contraction in a dose-dependent manner. ABL also showed a significant inhibitory effect on cell migration at 90 µg/ml, whilst 5-FU did not. A 10-min pulse of 5-FU exposure on ARPE-19 cells did not cause significant cell death.

Conclusions: : 5-FU and ABL regulate a number of the wound healing activities mediated by RPE cells.

Keywords: retinal pigment epithelium • wound healing • proliferative vitreoretinopathy