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The body excess weight of each rat was measured weekly, and the dosage of drug or vehicle administered was calculated based upon the most recent physique excess weight measurement.

The liver and left tibia Docetaxel of each animal were fixed with fixative and used for further histomorphometric analysis, while the right tibia was freed of all soft tissue and wrapped in a layer of PARAFILM, apart from 5 mm of its proximal end, and fixed into a 15 ml BD Falcon Tube and then soaked in fixative. The tube cap was tightened before performing a u CT scan to measure the microstructural parameters. The right femurs were subjected to DEXA measurement for BMD and bone mineral content. To assess bone loss, rats right tibiae were ex vivo scanned at the end of drug treatment. A 6 mm uCT scan with an isotropic resolution of 18 um was made of the proximal tibia using an in vivo u CT scanner The CT scanner was calibrated, and a beam hardening correction algorithm was applied to all scans. One CT scan took 35 minutes. In this study, the reproducibility E7080 of all structural parameters was high, with a coefficient of variation of about 1%.

With this protocol, the gray levels of voxels near the trabecular surfaces are not included to ensure that the measurements are not affected by partial volume effects. All DEXA measurements were Docetaxel performed by the same investigator using the Norland pDEXA Sabre equipped with Sabre Research software. The interassay coefficient of variation for BMD and BMC was 1. 7%. The scanner was calibrated daily to a dual material standard according to the manufacturers recommendations, and the scanner performance was controlled by the quality assurance protocol of our laboratory. The right femurs were scanned using DEXA to determine BMC and BMD.

Ex vivo measurements of the right distal femur were performed on excised bones positioned onto a 3 mmthick cotton piece on the bottom of a 10 cm diameter culture dish at a constant location on the scan table, and measured by DEXA using a special collimator, the scan length was 5 cm, the scan E7080 width 2 cm and the scan speed 10 mm/s with a resolution of 0. 2 mm ? 0. 2 mm.