Figure 3. NT-plasma induced osteogenic differentiation using qPCR markers.

(A) Fold increases in the expression of alkaline phosphatase (ALKP), bone morphogenetic protein 2 (BMP2), bone sialoprotein (BSP) and fibroblast growth factor-2 (FGF-2) in response to NT-plasma treatment, H₂O₂ or β-glycerol phosphate (βGP) normalized to sham treated control cells after 24 hr. ALKP was upregulated 3- (NS) and 11-fold (p<0.01), respectively, and BMP2 and BSP were increased 5 fold (p<0.05). βGP increased expression 14–23 fold (p<0.01). FGF-2 expression was not affected by NT-plasma, H₂O₂ or βGP. (B) After a 24 hr incubation in βGP NT-plasma was applied. 24 hr later there was a 2–7 fold increase in the induction of BMP2, ALKP (p<0.05) and Osterix was increased 15 fold (p<0.01) compared to βGP treatment alone. At 56 hr, BSP and osteocalcin (OSTCN) were increased 17–24 fold above βGP-treated control (p<0.05). The results are expressed as the mean ± standard deviation (n = 2). Statistical significance was determined by the Wilcoxin Mann-Whitney test for non-parametric data; * (p<0.05), ** (p<0.01).