Effect of Anti-inflammatory Mediator on the Proliferation of Human Corneal Keratocyte

ABSTRACT

PURPOSE: The purpose of this study was to evaluate the effect of anti-inflammatory mediators like dexamethasone, nordihyroguaratic acid (NDGA), and diclofenac sodium on proliferation of human corneal keratocytes, and to investigate the cellular morphology of keratocyte. METHODS: Human corneal keratocytes were exposed to 0.05, 0.1, 0.3, 0.8, and 1.0 mM concentration of each drug for period of 24, 48, and 72 hours. MTT based colorimetric assay was performed to assess the metabolic activity and inhibition of cellular proliferation. Cellular morphology was evaluated by inverted phase contrast micrograph and electron microscopy. RESULTS: The higher the concentration of inoculated each drugs was, the more the inhibitory effect of human keratocyte proliferation was found (P<0.05). NDGA, over 0.3 mM and diclofenac, more than 0.1 mM had significant more inhibitory effect on keratocyte proliferation compared with dexamethasone within 48 hours of exposure to each drug. With the concentration and exposure time of each drug, human corneal keratocytes were visible more rounded and swollen rather than spindle shape, and detached from the bottom of the dish. The damaged keratocytes had degenerative changes like cellular membrane disruption, microvilli disappearance, enlarged rough surfaced endoplasmic reticulum and mitochondria, vacuole formation and nuclear membrane damage by TEM. CONCLUSIONS: On basis of this study, the anti-inflammatory mediators such as NDGA and diclofenac sodium have less side effects and stronger inhibitory effects of human keratocyte proliferation than dexamethasone.