Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation.

Yu Wang,Haitao Cui,Peibiao Zhang,Zhenxu Wu,Xuesi Chen,Zongliang Wang,Yen Wei,Naipeng Wu,Jamshidkhan Chamani

doi:10.1371/journal.pone.0154924

Abstract

Electrical stimulation (ES) is therapeutic to many bone diseases, from promoting fracture regeneration to orthopedic intervention. The application of ES offers substantial therapeutic potential, while optimal ES parameters and the underlying mechanisms responsible for the positive clinical impact are poorly understood. In this study, we assembled an ES cell culture and monitoring device. Mc-3T3-E1 cells were subjected to different frequency to investigate the effect of osteogenesis. Cell proliferation, DNA synthesis, the mRNA levels of osteosis-related genes, the activity of alkaline phosphatase (ALP), and intracellular concentration of Ca2+ were thoroughly evaluated. We found that 100 Hz could up-regulate the mRNA levels of collagen I, collagen II and Runx2. On the contrary, ES could down-regulate the mRNA levels of osteopontin (OPN). ALP activity assay and Fast Blue RR salt stain showed that 100 Hz could accelerate cells differentiation. Compared to the control group, 100 Hz could promote cell proliferation. Furthermore, 1 Hz to 10 Hz could improve calcium deposition in the intracellular matrix. Overall, these results indicate that 100Hz ES exhibits superior potentialities in osteogenesis, which should be beneficial for the clinical applications of ES for the treatment of bone diseases.

Highlights

Electrical stimulation (ES) is clinically beneficial in the treatment of fracture, while 5%-10% of fractures show impaired healing and require additional orthopedic intervention
We find that cell proliferation is similar on 1 K and on control, while cell proliferation is decreased on 1, 10, 10 K, and 100 K compared to control at day 1
This study evaluates the role of ES frequency to determine which frequency most affects osteogenesis differentiation

Summary

Introduction

Electrical stimulation (ES) is clinically beneficial in the treatment of fracture, while 5%-10% of fractures show impaired healing and require additional orthopedic intervention. Fracture healing involves a complex multistep process. Serious periosteal and soft tissue damage at the time of fracture can lead to the formation of an atrophic nonunion. Fracture patients suffering from nonunion are mainly treated with surgery and orthopedic treatment, including bone grafts[1, 2], llizarov technique[3, 4], cytokines induction [5, 6], and biophysical therapy [7].

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