Stimulated myogenic differentiation of C2C12 murine myoblasts by using graphene oxide

Abstract

Currently, applications of graphene and its derivatives have been reported in many fields. In particular, graphene oxide (GO) has been studied for biomedical applications, such as cell imaging, biosensors and drug delivery systems, because of its good biocompatibility and excellent physicochemical properties. Recently, much research has been conducted to control cell behavior, including attachment, proliferation and differentiation by using GO. On the other hand, the effects of GO on myogenic differentiation have not been explored. In the present study, the influence of GO on myogenic differentiation was investigated. This study assessed the cytotoxicity of GO and evaluated the myogenic differentiation of C2C12 murine myoblasts in growth media containing 10 μg/mL of GO. The physicochemical properties of GO were characterized by using atomic force microscopy (AFM) and Raman spectroscopy. The cytotoxicity was evaluated by using a WST-8 assay. An analysis of myogenic differentiation was performed by using immunofluorescence staining for the myosin heavy chain (MHC; myogenic differentiation marker) and was determined by calculating the amount of multinucleate myotube formation, the fusion index and the maturation index. GO did not exhibit cytotoxicity at low concentrations and effectively stimulated the myogenic differentiation of C2C12 murine myoblasts. Therefore, GO has potential skeletal tissue engineering applications.