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. Author manuscript; available in PMC: 2018 Sep 15.
Published in final edited form as: Acta Biomater. 2017 Jul 12;60:181–189. doi: 10.1016/j.actbio.2017.07.017

Figure 3. Down-regulation of SHED osteogenesis in the presence of pro-inflammatory cytokines.

Figure 3

(a) Alizarin red staining showed reduced mineralized nodule formation for SHED cultured in osteogenic media in the presence of IFN-γ 50 (ng m/l)- TNF-α (5 ng m/L) combination. Mineralization area percentage was defined as the area of stained mineralization divided by the total area. (b) The protective role of the biomaterial was confirmed by osteogenic differentiation of encapsulated SHED in alginate with different degrees of elasticity/porosity in the presence of IFN-γ and TNF-α for 4 weeks and staining with Xylenol orange fluoroprobe. (c) Mineralization area percentage was defined as the area of stained mineralization divided by the total area of the field of view of the image in d. (d) Western blot analysis showed that pro-inflammatory cytokine-treated SHED expressed decreased levels of osteogenic marker. Moreover, Western blot analysis confirmed that the physiomechanical properties of the encapsulating hydrogel biomaterial regulated the expression of osteogenic marker levels (RUNX2) in the presence of IFN-γ and TNF-α. NS= not significant, *P<0.05, **P<0.01.