Figure 3

Odonto/osteogenic differentiation was enhanced in E2-treated SCAP. (A) Immunocytochemical staining of STRO-1 in the control group. (B) Immunocytochemical staining of STRO-1 in the E2 group. Scale bars = 100 μm. (C) ALP activity in the control group, E2 group, mineralization-inducing group (MM) and MM + E2 group at day 5 and 7, respectively. Values are presented as the mean ± SD, n = 3. **P <0.01. (D) Alizarin red staining for the control group, E2 group, MM group and MM + E2 group at day 14. (E) Quantitative analysis of calcium content in the different groups. Values are the mean ± SD, n = 3. *P <0.05. (F-I) Calcium nodules in the different groups under the inverted microscope. Scale bars = 100 μm. (J) Real-time RT-PCR analysis for the expression of ALP, DMP1, DSPP, RUNX2, OSX and OCN in control and E2-treated SCAP at day 3. Values are the mean ± SD, n = 3. **2^-ΔΔCt >2, P <0.01; *1 < 2^-ΔΔCt <2, P <0.01. (K) Real-time RT-PCR analyses for the expression of ALP, DMP1, DSPP, RUNX2, OSX and OCN in control and E2-treated SCAP at day 7. Values are the mean ± SD, n = 3. **2^-ΔΔCt >2, P <0.01; *1 < 2^-ΔΔCt <2, P <0.01. (L) Western blot assay for the odonto/osteogenic proteins (DMP1, DSP, RUNX2, OSX and OCN) in control and E2-treated SCAP at day 3. (M) Quantitative analysis of western blot results at day 3. Values are the mean ± SD, n = 3. **P <0.01. (N) Western blot assay of the odonto/osteogenic proteins in control and E2-treated SCAP at day 7. (O) Quantitative analysis of western blot results at day 7. Values are the mean ± SD, n = 3. *P <0.05, **P <0.01. ALP, alkaline phosphatase; E2, 17beta estradiol; SCAP, stem cells from apical papilla; SD, standard deviation.