The Study of the Effects of Glucose on Cultured Human Corneal Epithelial Cells

ABSTRACT

PURPOSE: In this study, we evaluated the effects of glucose on the proliferation, migration, morphological characteristics, and proteinase expressions in human corneal epithelial cells to discover the cause of diabetic corneal epithelial complications. METHODS: Human corneal epithelial cells transfected by SV40 were cultured in media containing 17.5 mM (control), 25 mM, and 100 mM D-glucose to stimulate a diabetic condition. In some experiments, 25 mM or 100mM D-mannitol was added to the media to control for the osmotic effect of glucose. We performed MTT analysis of the proliferation assay, cell count of the migration assay, scanning electron microscopic (SEM) examination for morphologic change, and RT-PCR for the expression of mRNA of metalloproteinase (MMP). RESULTS: Raising the concentration of glucose from 25 mM to 100 mM significantly decreased in the cellular proliferation and migration. These findings were not observed in the presence of 25 mM or 100 mM mannitol. In SEM, the pseudopods of the cells were markedly decreased in the 100mM glucose media. Although the expression of MMP-2 was not significantly different, that of MMP-9 was significantly increased with increasing glucose concentration. CONCLUSIONS: Higher levels of glucose show a significant effect on cellular proliferation, migration, morphology, and the expression of MMP-9 in corneal epithelial cells. This study may be an initial step in understanding the mechanism of corneal epithelial wound healing in diabetic patients.