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. 2019 Dec 27;9:19895. doi: 10.1038/s41598-019-56151-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of long-term administrations of anti-RANKL antibody or ZOL on femur and tibia. (A) Whole femur appearance. (B) Micro-computed tomography (μCT) findings of distal femurs in 8-week-old mice administered saline, anti-RANKL antibody (Ab) at 2.5 mg/kg, RfD-ZOL at 0.08 mg/kg, or CD-ZOL at 3.0 mg/kg weekly for 7 weeks. (C) Findings following toluidine blue staining (upper) and TRAP staining (middle and lower) of proximal tibiae growth plate obtained from 8-week-old mice after weekly administrations of each drug for 7 weeks. Higher magnification images of black-boxed regions in middle panels are shown in lower panels. Arrows indicate TRAP-positive cells. Representative results are shown in (A–C). (D) Femur bone length. (E) Bone volume/tissue volume (BV/TV) ratios were determined by μCT (saline, n = 6; anti-RANKL Ab, n = 5; Rfd-ZOL, n = 6, CD-ZOL, n = 5). (F) Number of osteoclasts (N.Oc) was determined in proximal tibiae following TRAP staining. Four mice from each of the saline, anti-RANKL Ab, RfD-ZOL, and CD-ZOL groups were used. Statistical differences were assessed by one-way ANOVA with Tukey-Kramer’s test. ^†Statistically significant different from compared to indicated groups, p < 0.05. Error bars represent SEM.