Bone samples were fixed in 4% formaldehyde.
After micro-CT scans, the extracted bone samples were fixed in a neutral 10% formaldehyde solution, and the tissue blocks were embedded in a resin for histological analysis.
Bone samples were fixed in 10% neutral buffered formalin, then decalcified in 4% ethylenediamine tetraacetic acid (pH 7.2) (Sigma-Aldrich, St Louis, MO, USA).
All bone samples were fixed in 40% ethanol for 1 week, followed by a series of ethanol dehydration (50-100%) within 6 days.
For histology, trabecular bone samples were fixed in 70% ethanol, processed undecalcified through a graded series of ethanol, embedded in methylmethacrylate resin, and sectioned on a microtome (Polycut-E, Leica SP2600; Leica Microsystems, Wetzlar, Germany), as previously described [ 24, 31].
Biopsies were taken from the iliac crest of adult sheep (∼10 years old; approved by Danish Animal Experiments and Inspectorates, 2011/561-1959) from the study described in Andreasen et al. The bone samples were fixed in 4% buffered formaldehyde for 24 h, before transferring to 0.1% buffered formaldehyde.
Bone tissue samples were fixed in 4% paraformaldehyde overnight and decalcified in 10% EDTA for 10 days.
Briefly, bone marrow samples were fixed in formalin for 24 h, decalcified, embedded in paraffin, and sliced.
Bone and spleen samples were fixed in 10% formalin, washed with PBS and stored to 80% ethanol.
The bone and synovial membrane samples were fixed in 4% paraformaldehyde at 4°C and further processed for histology.
