Real-time RT-PCR analysis.

NPWT-treated and static control group cell clots were harvested before and 1, 3, and 7 days after incubation (n = 3 per timepoint for each group) and analyzed by real-time RT-PCR. The expressions of the specific osteogenic genes (A) ALP, (B) OC, and (D) COL1 were higher in NPWT group than those in control group (*p<0.05, **p<0.01). The NPWT group also expressed higher levels of the osteogenic transcription factor Cbfa1 (also known as Runx2) and the mechanosignaling molecule integrin β5 (ITGB5) (C) and (E) (*p<0.05, **p<0.01).