Abstract
Electrical signals represent an essential form of cellular communication. For decades, electrical stimulation has been used effectively in clinical practice to enhance bone healing. However, the detailed mechanisms between electrical stimulation and bone healing are not well understood. In addition, there have been many difficulties in setting up a stable and efficient electrical stimulation system within the in vitro environment. Therefore, various conductive materials and electrical stimulation methods have been tested to establish an effective electrical stimulation system. Through these systems, many studies have been conducted on the effects of electrical stimulation on bone healing and osteogenic differentiation. However, previous studies were limited by the use of opaque conductive materials that obscure the cells; fluorescent observations and staining are known to be two of the critical methods to confirm the states of the cells. Indium tin oxide (ITO) glass is known to have excellent transparency and conductivity, but it is challenging to cultivate cells due to low cell adhesion characteristics. Therefore, we used O2 plasma treatment to increase the hydrophilicity and wettability of ITO glass. This enhanced cell affinity to the glass, providing a stable surface for the cells to attach. Then, electrical stimulation was applied with an amplitude range of 10 to 200 µA at a frequency of 10 Hz. Our results demonstrated that the osteogenic differentiation efficiency was maximized under the amplitude conditions of 10 µA and 50 µA. Accordingly, the results of our study suggest the development of an excellent platform in the field of biological research as a good tool to elucidate various mechanisms of cell bioactivity under electrical conditions.