Yunnan Baiyao Conditioned Medium Promotes the Odonto/Osteogenic Capacity of Stem Cells from Apical Papilla via Nuclear Factor Kappa B Signaling Pathway

Figure 4

Effects of cotreatment by YNB and NF-κB inhibitor BMS345541 on the odonto/osteogenic differentiation of SCAPs. (a) The protein levels of cytoplasm IκBα, p-IκBα, P65, and p-P65 in YNB-treated SCAPs at different time points. GAPDH served as a loading control. (b-e) Semiquantitative analyses of cytoplasmic IκBα, p-IκBα, P65, and p-P65. n=3. P<0.05, P<0.01. (f, g) Immunofluorescence staining of P65 and IκBα in YNB-CM treated SCAPs at 0, 15, 30, 60, 120 min and with NF-κB inhibitor BMS345541, respectively. Scale bar = 200 μm. (h, i) ALP activity and ALP staining in BMS345541 + YNB group and YNB group after 3 days of culture. Values are means ± SD, n=3. P<0.05, P<0.01. (j, k) mRNA expression of OCN, OPN, OSX, RUNX2, ALP, COL-I, and DSP in different groups by real-time RT-PCR after cotreatment with YNB and BMS345541 at day 3 and day 7, respectively. , n=3. (l, o) The expression levels of the odonto/osteogenic proteins in different groups after cotreatment with BMS345541 and YNB at day 3 and day 7, respectively. Values are means ± SD, n=3. P<0.05, P<0.01. (m, n) Grayscale analyses of the odonto/osteogenic proteins at day 3. P<0.05, P<0.01. (p, q) Grayscale analyses of the odonto/osteogenic proteins at day 7. P<0.05, P<0.01.