Pulsed electromagnetic field induces Ca2+-dependent osteoblastogenesis in C3H10T1/2 mesenchymal cells through the Wnt-Ca2+/Wnt-β-catenin signaling pathway

Abstract

Pulsed electromagnetic fields (PEMFs) are effective in healing fractures and improving osteoporosis. However, their effect on mesenchymal cells remains largely unknown. In this study, the effects of PEMF on osteoblastogenesis and its underlying molecular signaling mechanisms were systematically investigated in C3H10T1/2 cells. C3H10T1/2 mesenchymal cells were exposed to 30-Hz PEMF bursts at various intensities for 3 consecutive days. The optimal PEMF exposure (30 Hz, 1 mT, 2 h/day) was applied in subsequent experiments. Our results suggest that intracellular [Ca²⁺]i in C3H10T1/2 cells can be upregulated upon exposure to PEMF and that PEMF-induced C3H10T1/2 cell differentiation was Ca²⁺-dependent. The pro-osteogenic effect of PEMF on Ca²⁺-dependent osteoblast differentiation was then verified by alkaline phosphatase (ALP) and von Kossa staining. Furthermore, PEMF promoted the gene expression and protein synthesis of the Wnt/β-catenin pathway. Increased [Ca²⁺]i in the nucleoplasm was followed by the mobilization and translocation of β-catenin into the nucleus in C3H10T1/2 cells. A model of Wnt/β-catenin signaling and the Wnt/Ca2+ signaling network is proposed. Taken together, these findings indicated for the first time that PEMF induces osteoblastogenesis through increased intracellular [Ca²⁺]i and the Wnt-Ca2+/Wnt-β-catenin signaling pathway in C3H10T1/2 mesenchymal cells.

Keywords: Ca(2+) upregulation; Osteoblastogenesis; PEMF; Wnt-β-catenin.