Heliyon (Jul 2023)
X-ray diffraction studies of a partially demineralized oriented cortical bone with the controlled depth of analysis
Abstract
The in vitro demineralization of bone tissue is used for simulating the osteoporosis related bone loss. This way would be helpful in observations of bone apatite dissolution in microstructural level and may give significant input for understanding crystal-chemistry of bone resorption. In the case of cortical bone, demineralization occurs inhomogeneously, with the formation of a superficial demineralized layer and a transition zone with a gradient of concentration and structural characteristics perpendicular to the reaction advance front. Changes in the microstructural parameters of the bone mineral in this interface zone are of great interest for understanding the resorptive processes in the bone associated with osteoporosis. In this work, the SEM-EDX method was used to estimate the sizes of the demineralized and interface layers in the cortical bone during stepwise demineralization in HCl water solution; the general patterns of changes in the concentrations of Ca, P, and Cl in these layers were established. The calculations of the effective penetration depth of X-rays in diffraction mode for the intact and partially demineralized cortical bone were performed. It is shown that the use of CoKα radiation (instead of the usual CuKα) ensures the depth of probing within the interface zone, which allows to adequately assess the microstructural parameters (crystallite sizes and lattice microdeformations) of altered bioapatite in the zone of its interaction with an acid agent. A nonmonotonic change in the average size of crystallites and microdeformations of the apatite lattice was revealed during acid demineralization of the bone. Using asymmetric XRD geometry, the evidence was obtained that the affected mineral of the transition zone does not contain other crystalline phases except for weakly crystallized apatite. For the first time, the depth-controlled XRD analysis was applied to such a complex (surface-gradient) object as partially demineralized cortical bone. Additionally, we propose a rapid, averaging, and non-destructive method for estimating the depth of the reaction front dividing the demineralized and non-demineralized portions of the bone by XRD. The consistency of XRD and SEM-EDX data on the thickness values of the demineralized layer is shown.
