SHED transplantation blocked early osteoporotic phenotype development in OVX mice. (A) Schema indicating the experimental design for SHED or hBMMSC transplantation in OVX mice. (B) μCT images of trabecular bone structure in the femurs of sham, OVX, SHED-transplanted, and hBMMSC-transplanted mice. OVX mice showed a reduced trabecular bone volume compared with that in the sham group. SHED and hBMMSC transplantation partially rescued OVX-induced reduction of trabecular bone volume. (C-I) μCT analysis showed that OVX induced reduced bone volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), bone mineral density (BMD), and connectivity density (Conn.D), along with increased trabecular space (Tb.Sp) and structure model index (SMI) in the femurs. (J) μCT analysis showed that the cortical bone volume in the femur was reduced in OVX mice. SHED and hBMMSC transplantation increased the cortical bone volume in the femurs of OVX mice. (K-N) μCT analysis showed that OVX resulted in reduced total cross-sectional area (Tt.Ar), cortical bone area (Ct.Ar), cortical bone fraction (Ct.Ar/Tt.Ar), and cortical thickness (Ct.Th) in the femurs of OVX mice. SHED and hBMMSC transplantation significantly increased Tt.Ar, Ct.Ar, Ct.Ar/Tt.Ar, and Ct.Th in OVX mice. (O) Hematoxylin and eosin (H&E) staining showed that OVX resulted in reduced trabecular bone area (yellow circled area) compared with the sham mice, as analyzed by ImageJ software. SHED and hBMMSC transplantation significantly increased the trabecular bone area. (P) TRAP staining showed that OVX resulted in increased TRAP+ cells (yellow arrows) in the femurs of OVX mice. SHED and hBMMSC transplantation reduced the number of TRAP+ cells in the femurs of OVX mice. (Q-R) The levels of CTX-1, TRAP 5b, and RANKL in serum were markedly decreased in both SHED and hBMMSC transplantation groups compared with the “OVX without transplantation” group, whereas OPG levels were markedly increased in both treatment groups. n = 5 in each group. Scale bar = 200 μm; *p < .05; **p < .01; Error bars: mean ± SD.