Histological Analysis of Vertebral Morphology

Sections used for histology were fixed in 4 % paraformaldehyde (PFA) (VWR, Lutterworth, UK) for 20 min, stained with 0.025 % alcian blue (Merck) in 3 % acetic acid (Merck) (for cartilage) for 30 min followed by 1 % picrosirius red (Merck) (for collagen) for 30 min (Fig. 1c) (Rolfe et al., 2017 (link)). Sections from the cervical, thoracic, and lumbar regions were imaged in transmitted illumination using a light microscope (Yenway EX30, Glasgow, UK), with light intensity being adjusted to observe the vertebral bodies. Vertebral segmentation, disc morphology, and vertebral shape were qualitatively compared between control and mdg spines, using light microscopy images.