Acellular calcified columns in the normal growth plate of mouse vertebrae

The present study aims to demonstrate the calcified columnar structures of the growth plate of mouse vertebrae and to show their age related changes. For light microscopy, paraffin sections of decalcified lumbar spines were stained with hematoxylin-eosin or toluidine blue; methacrylate sections of undecalcified specimens were stained to detect calcium precipitate. For scanning electron microscopy, lumbar spines treated with 5% NaClO solution were dehydrated by acetone and metal-coated. Light microscopy of hematoxylin-eosin stained sections revealed that the acellular columnar structures appeared between the chondrocyte stacks in the growth plate. These structures were stained more densely by toluidine blue. The methacrylate-embedded sections for calcium staining showed calcium deposition in the columns. Scanning electron microscopy of NaClO-treated specimens enabled the direct observation of the columns on both the epiphyseal and diaphyseal bone surfaces facing the cartilaginous growth plate. Numerous projections on each surface were distributed in mirror images ; the corresponding projections were similar in size and shape, indicating that the projections bonded with each other and formed calcified columns in the cartilaginous growth plate. Longitudinal sections of the spine confirmed these findings. The calcified columns first appeared about 2 or 3 weeks of age and increased in number with time. While increasing in number, they also grew in size fusing with the neighboring ones. The proportional area of the columns occupying the surface facing the growth plate also increased with age. These findings indicate that the calcified column ultimately concerns the cessation of the bone growth.