Fig. 6

Effects of LY294402 and rapamycin on in vitro calcification of scaffold-free SCAP-based spheroids. SCAP-based spheroids were pretreated with or without LY294402 (50 μM) and rapamycin (100 nM) and were cultured under osteogenic/dentinogenic condition for 4 weeks. a–c Representative microscopic images (a), 3D (micro CT-3D, b) and tomographic (microCT-2D, c) micro-CT images of SCAP-based calcified spherical constructs. d, e Micro-CT analysis shows bone parameters (BV/TV, ratio of bone volume [BV] per total volume [TV]; Tb.Th, trabecular thickness; Tb.N, trabecular numbers; Tb.Sp, trabecular separation) (d) and bone mineral density (BMD) (e). f RT-qPCR analysis shows the expression of RUNX2 and DSPP in SCAP-transplant tissues. Results show the ratios to the non-pretreated SCAP-transplant group. d–f: n = 5 for all groups. *P < 0.05, **P < 0.01 and ***P < 0.005. Graph bars show the means ± SEM. a–c, f Intact, non-pretreated SCAP-transplanted group