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研究目的（著者による）
To study the effects of different waveforms on cell proliferation, differentiation and mineralization of rat osteoblasts.
詳細情報
Four different waveforms and an untreated control were examined: sinusoidal, triangular, rectangular and sawtooth. Osteoblasts (primary cells) were isolated from skull bones of neonatal rats.
影響評価項目
cell proliferation, cell differentiation and mineralization potentials of osteoblasts
osteoblasts placed in 60 mm culture dishes inside the solenoid
ばく露装置の詳細
solenoid consisting of three series-connected coils and the core cylinder was an acrylic tube (inner diameter = 18 cm, height = 21 cm); solenoid positioned in an incubator at 37°C
分子生合成: gene expression of different osteogenesis-related genes after 1, 3 and 4 days of exposure (Igf-1, Opg, Runx-2; real-time RT-PCR)
細胞生存力／細胞分裂/増殖: cell proliferation after 3 days of exposure (MTT assay)
cell differentiation after 3, 6, 9 and 12 days of exposure: osteoblast mineralization (amount of calcium, numbers and area of calcified nodules; Alizarin red staining), alkaline phosphatase (ALP) enzyme activity (as early marker of osteoblast maturation phase; ALP positive colonies; commercial kit and histochemical staining (after 8 days of exposure)) and gene expression (see "molecular biosynthesis")
Compared to the untreated control, sinusoidal magnetic field exposure significantly inhibited osteoblast proliferation while rectangular magnetic field exposure significantly increased the proliferation. After 9 and 12 days of exposure, the enzyme activity of the alkaline phosphatase was significantly higher in the sinusoidal and triangular waveform exposed cultures compared to the control cultures. The amount of alkaline phosphatase positive colonies was significantly higher in the sinusoidal, triangular and sawtooth waveform exposed cultures than in the control cultures. Regarding mineralization, after 9 and 12 days of exposure to sinusoidal and triangular waveforms (and amount of calcium also sawtooth after 12 days), the amount of calcium and the number of calcified nodules were significantly higher than in the control cultures. In comparison to the control cultures, the gene expression of Runx-2 and Opg was significantly upregulated after 1 day of exposure to a sinusoidal, triangular and sawtooth waveform, whereas the gene expression of Igf-1 was significantly increased after 3 and 4 days. While rectangular magnetic fields increased the proliferation and did not effect the differentiation of osteoblasts, sinusoidal fields inhibited the proliferation but promoted osteogenic differentiation. The authors conclude that the influence of magnetic fields on proliferation, differentiation and mineralization of rat osteoblasts is dependent on the waveform.
