Effect of Glucose on Retinal Endothelial Cell Viability and VEGF Secretion.

Previous studies have shown that in diabetic patients, there is an increase of retinal capillaries associated with the development of diabetic retinopathy in the eye. The objective of current study is to investigate the effect of glucose on retinal endothelial cell viability and VEGF secretion. 20,000 cells per well were treated without glucose or with 5.5mM (euglycemic), 18.5mM and 30mM (hyperglycemic) glucose for 24 hours. Viable cells were counted using Trypan blue dye exclusion method. ELISA was used to measure VEGF secretion from cells into the cell medium. The number of viable cells incubated with 5.5mM glucose (physiological control) increased by 53.7% after 24 hours. In comparison, cells treated with 18.5mM glucose decreased by 2.8% while cells treated with 30mM glucose decreased by 20% after 24 hours of incubation. Cells without glucose treatment (0mM control) decreased by 33.3%. In contrast to the decrease of viable cell numbers after treatment with high glucose, there is an increase in VEGF secretion (pg/mL) to the cell medium with increase in glucose concentration from 5.5mM to 0, 18.5, and 30mM. The amount of VEGF secreted per cell also increased with increasing glucose concentrations. Our results show that viability of retinal endothelial cells and VEGF release are highly responsive to changes in glucose concentration. Such glucose-induced changes in retinal endothelial cells may negatively impact the integrity of the microvasculature in the diabetic retina leading to angiogenesis and microaneursyms.