Intracortical stiffness of mid-diaphysis femur bovine bone: lacunar-canalicular based homogenization numerical solutions and microhardness measurements.

Microscale lacunar-canalicular (L-C) porosity is a major contributor to intracortical bone stiffness variability. In this work, such variability is investigated experimentally using micro hardness indentation tests and numerically using a homogenization scheme. Cross sectional rings of cortical bones are cut from the middle tubular part of bovine femur long bone at mid-diaphysis. A series of light microscopy images are taken along a line emanating from the cross-section center starting from the ring's interior (endosteum) ring surface toward the ring's exterior (periosteum) ring surface. For each image in the line, computer vision analysis of porosity is conducted employing an image segmentation methodology based on pulse coupled neural networks (PCNN) recently developed by the authors. Determined are size and shape of each of the lacunar-canalicular (L-C) cortical micro constituents: lacunae, canaliculi, and Haversian canals. Consequently, it was possible to segment and quantify the geometrical attributes of all individual segmented pores leading to accurate determination of derived geometrical measures such as L-C cortical pores' total porosity (pore volume fraction), (elliptical) aspect ratio, orientation, location, and number of pores in secondary and primary osteons. Porosity was found to be unevenly (but linearly) distributed along the interior and exterior regions of the intracortical bone. The segmented L-C porosity data is passed to a numerical microscale-based homogenization scheme, also recently developed by the authors, that analyses a composite made up of lamella matrix punctuated by multi-inclusions and returns corresponding values for longitudinal and transverse Young's modulus (matrix stiffness) for these micro-sized spatial locations. Hence, intracortical stiffness variability is numerically quantified using a combination of computer vision program and numerical homogenization code. These numerically found stiffness values of the homogenization solution are corroborated experimentally using microhardness indentation measurements taken at the same points that the digital images were taken along a radial distance emanating from the interior (endosteum) surface toward the bone's exterior (periosteum) surface. Good agreement was found between numerically calculated and indentation measured stiffness of Intracortical lamellae. Both indentation measurements and numerical solutions of matrix stiffness showed increasing linear trend of compressive longitudinal modulus (E11) values vs. radial position for both interior and exterior regions. In the interior (exterior) region of cortical bone, stiffness modulus values were found to range from 18.5 to 23.4 GPa (23 to 26.0 GPa) with the aggregate stiffness of the cortical lamella in the exterior region being 12% stiffer than that in the interior region. In order to further validate these findings, experimental and FEM simulation of a mid-diaphysis bone ring under compression is employed. The FEM numerical deflections employed nine concentric regions across the thickness with graded stiffness values based on the digital segmentation and homogenization scheme. Bone ring deflections are found to agree well with measured deformations of the compression bone ring.