Effect of TNF-α-Induced Sclerostin on Osteocytes during Orthodontic Tooth Movement

Figure 2

Tumor necrosis factor alpha (TNF-α) enhanced sclerostin expression in primary osteocytes, and sclerostin enhanced osteoclastogenesis in vitro. (a) Expression of SOST mRNA in osteocytes, analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). Total RNA in osteocytes was isolated from osteocytes cultured with or without TNF-α for 1 or 3 days (; ). (b) Expression of RANKL mRNA in osteocytes, analyzed by real-time RT-PCR. Total RNA from osteocytes was isolated from osteocytes cultured with recombinant human sclerostin (rhSCL) (0 ng/mL, 1 ng/mL, 10 ng/mL, and 100 ng/mL) for 2 days (; ). (c) Expression levels of RANKL and OPG mRNA, as well as RANKL/OPG ratio in osteocytes, analyzed by real-time RT-PCR. Total RNA was isolated from osteocytes cultured with or without rhSCL (100 ng/mL) for 2 days (; and ). (d) Microscopic photos and the numbers of tartrate-resistant acid phosphatase- (TRAP-) positive cells within a coculture of osteocytes and osteoclast precursors, with or without rhSCL, in the presence of vitamin D3 and prostaglandin E2. (; ).

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