Rapid alterations of avian medullary bone material during the daily egg-laying cycle.

Bone is a dynamic tissue which is continuously adapting not only to external mechanical stimuli but also to internal metabolic calcium demands. During normal bone remodeling, bone-resorbing osteoclasts release calcium from the bone and digest the collagenous bone matrix, after which bone-depositing osteoblasts form unmineralized collagen matrix, which subsequently mineralizes. The detailed mechanism by which calcium is deposited at the site of mineralization and removed from it during bone resorption is largely unknown. Experimental studies are difficult to conduct because in adult bone only a small fraction of bone tissue is remodeled at any moment in time. Thus, one promising approach is to study mineral deposition and resorption in model systems in which a large fraction of the bone mineral is mobilized in a relatively short period of time. We investigated the microscopic and nanoscopic alterations of avian medullary bone architecture during the egg-laying (oviposition) cycle of hens. Medullary bone forms a labile calcium reservoir for eggshell production and is characterized by an extremely rapid and high-flux calcium metabolism. It thus, provides the unique opportunity to study processes of bone remodeling in their most intensive form. We used a combination of synchrotron X-ray tomography together with small angle X-ray scattering (SAXS), wide angle X-ray diffraction (WAXD) and X-ray fluorescence (XRF) to correlate microscopic medullary bone attributes such as the mineral content, medullary bone volume fraction and medullary bone trabecular thickness with nanoscopic alterations in the mineral particle size (thickness parameter T and length parameter L) during the oviposition cycle. To identify the timing of the different stages of the cycle, ionic calcium, phosphorus and PTH concentrations in the blood of the layers were monitored. We found that the microscopic and nanoscopic architecture of avian medullary bone material changes rapidly during the oviposition cycle. During eggshell calcification, the mineral content and the size of trabeculae of medullary bone decrease markedly. Furthermore, the average mineral particle size increases during resorption, suggesting that the smaller mineral particles are preferrentially removed. Medullary bone thus formes a fast-responding system exhibiting rapid alterations of the material at the micron and nano scale. Those mechanisms are crucial to provide calcium for the high metabolic calcium demand during eggshell mineralization.