Study On Safety And The Anti-oxidative Stress Effects Of The Fullerene Derivatives Towards Corneal Epithelium

Purpos:As a brand new nanomaterial,Fullerene is shaped by 60 carbon atoms with a diameter of approximately 0.71nm.Recently,they were extensively studied in order to apply in bio-medicine field because of their strong anti-oxidative abilities.However,little is known about their biosafety and their potential pharmacological effects in cornea.In this study we used water soluble fullerene derivatives C60(COOH)12 to investigated the biosafety and potential anti-oxidative stress effects of C60(COOH)12 on HCE cells and in rats.Methods:First we investigated the toxic effects of C60(COOH)12.The following experiments were performed:CCK-assay to detect cell viability,clinical slip lamp to observe general changes and histological staining were conducted after topical treatment on rat cornea for one week.In the anti-oxidative stress experimets,we applied hydrogen peroxide(H2O2)induced oxidative stress model in the corneas of Wistar rat.The levels of main factors of oxidative stress 3-nitrotyrosine(3-NT)and Keapl-Nrf2-ARE signaling pathway were assessed by western blot and immunofluorescence staining.We also applied H2O2-induced models in cultured HCE cells.CCK-8 assay,Western blot and immunofluorescence staining were performed to detect the level of correlation components of ROS system.Conclusion:Fullerene derivatives C60(COOH)12 did not have toxic effects as in HCE cell in vitro and the rat cornea in vivo;fullerene derivatives protected the oxidant damage induced by hydrogen peroxide(H2O2)both in HCE cells and in rat cornea,the underlying mechsnism is via the activation of Keap l-Nrf2-ARE signaling pathway.