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. 2020 Jun 12;10(16):7409–7421. doi: 10.7150/thno.44782

Figure 3.

Figure 3

HERS spheroids, rather than 2D monolayer HERS cells, maintained the cementogenesis potential both in vitro and in vivo. (A) The alizarin red staining of mineralized deposits formed by 2D monolayer HERS cells (upper) and HERS spheroids (lower) indicated that more mineralized deposits formed in the HERS spheroids groups after induction. (B) Light microscope images of alizarin red staining also revealed that more mineralized deposits formed in the HERS spheroids groups, also. (C) Quantification of the alizarin red staining showed a consistent trend. (D) RT-qPCR analysis of the relative expression of cementogenesis-associated genes BSP and DMP1 after induction. Their relative expression level is higher in the HERS spheroids groups. (E) Cementogenesis potential was compared in vivo. H&E staining revealed a thin layer of cementum-like tissue generated on the surface of HA/TCP particles in the HERS spheroids groups, but not in the 2D monolayer HERS cells transplant (n=4). IHC showed the cementum-like tissues were positive for DMP1, BSP, and OCN antibody compared to the nearly-negative stain in the 2D monolayer HERS cells transplants. Scale bars are shown, *** P < 0.001; ** P< 0.01; * P < 0.05.