Ultrasounds could be considered as a future tool for probing growing bone properties.

Juvenile bone growth is well described (physiological and anatomical) but there are still lacks of knowledge on intrinsic material properties. Our group has already published, on different samples, several studies on the assessment of intrinsic material properties of juvenile bone compared to material properties of adult bone. The purpose of this study was finally to combine different experimental modalities available (ultrasonic measurement, micro-Computed Tomography analysis, mechanical compression tests and biochemical measurements) applied on small cubic bone samples in order to gain insight into the multiparametric evaluation of bone quality. Differences were found between juvenile and adult groups in term of architectural parameters (Porosity Separation), Tissue Mineral Density (TMD), diagonal stiffness coefficients (C33, C44, C55, C66) and ratio between immature and mature cross-links (CX). Diagonal stiffness coefficients are more representative of the microstructural and biochemical parameters of child bone than of adult bone. We also found that compression modulus E was highly correlated with several microstructure parameters and CX in children group while it was not at all correlated in the adult group. Similar results were found for the CX which was linked to several microstructure parameters (TMD and E) only in the juvenile group. To our knowledge, this is the first time that, on a same sample, ultrasonic measurements have been combined with the assessment of mechanical and biochemical properties. It appears that ultrasonic measurements can provide relevant indicators of child bone quality (microstructural and biochemical parameters) which is promising for clinical application since, B-mode ultrasound is the preferred first-line modality over other more constraining imaging modalities (radiation, parent-child accessibility and access to the patient's bed) for pediatric patients.

Compositional and mechanical properties of growing cortical bone tissue: a study of the human fibula.

Human cortical bone contains two types of tissue: osteonal and interstitial tissue. Growing bone is not well-known in terms of its intrinsic material properties. To date, distinctions between the mechanical properties of osteonal and interstitial regions have not been investigated in juvenile bone and compared to adult bone in a combined dataset. In this work, cortical bone samples obtained from fibulae of 13 juveniles patients (4 to 18 years old) during corrective surgery and from 17 adult donors (50 to 95 years old) were analyzed. Microindentation was used to assess the mechanical properties of the extracellular matrix, quantitative microradiography was used to measure the degree of bone mineralization (DMB), and Fourier transform infrared microspectroscopy was used to evaluate the physicochemical modifications of bone composition (organic versus mineral matrix). Juvenile and adult osteonal and interstitial regions were analyzed for DMB, crystallinity, mineral to organic matrix ratio, mineral maturity, collagen maturity, carbonation, indentation modulus, indicators of yield strain and tissue ductility using a mixed model. We found that the intrinsic properties of the juvenile bone were not all inferior to those of the adult bone. Mechanical properties were also differently explained in juvenile and adult groups. The study shows that different intrinsic properties should be used in case of juvenile bone investigation.

Pore network microarchitecture influences human cortical bone elasticity during growth and aging.

Cortical porosity is a major determinant of bone strength. Haversian and Volkmann׳s canals are׳seen' as pores in 2D cross-section but fashion a dynamic network of interconnected channels in 3D, a quantifiable footprint of intracortical remodeling. Given the changes in bone remodeling across life, we hypothesized that the 3D microarchitecture of the cortical pore network influences its stiffness during growth and ageing. Cubes of cortical bone of 2 mm side-length were harvested in the distal 1/3 of the fibula in 13 growing children (mean age±SD: 13±4 yrs) and 16 adults (age: 75±13 yrs). The cubes were imaged using desktop micro-CT (8.14µm isotropic voxel size). Pores were segmented as a solid to assess pore volume fraction, number, diameter, separation, connectivity and structure model index. Elastic coefficients were derived from measurements of ultrasonic bulk compression and shear wave velocities and apparent mass density. The pore volume fraction did not significantly differ between children and adults but originates from different microarchitectural patterns. Compared to children, adults had 42% (p=0.033) higher pore number that were more connected (Connective Density: +205%, p=0.001) with a 18% (p=0.007) lower pore separation. After accounting for the contribution of pore volume fraction, axial elasticity in traction-compression mode was significantly correlated with better connectivity in growing children and with pore separation among adults. The changes in intracortical remodeling across life alter the distribution, size and connectedness of the channels from which cortical void fraction originates. These alterations in pore network microarchitecture participate in changes in compressive and shear mechanical behavior, partly in a porosity-independent manner. The assessment of pore volume fraction (i.e., porosity) provides only a limited understanding of the role of cortical void volume fraction in its mechanical properties.

Measuring mass density and ultrasonic wave velocity: A wavelet-based method applied in ultrasonic reflection mode.

When assessing ultrasonic measurements of material parameters, the signal processing is an important part of the inverse problem. Measurements of thickness, ultrasonic wave velocity and mass density are required for such assessments. This study investigates the feasibility and the robustness of a wavelet-based processing (WBP) method based on a Jaffard-Meyer algorithm for calculating these parameters simultaneously and independently, using one single ultrasonic signal in the reflection mode. The appropriate transmitted incident wave, correlated with the mathematical properties of the wavelet decomposition, was determined using a adapted identification procedure to build a mathematically equivalent model for the electro-acoustic system. The method was tested on three groups of samples (polyurethane resin, bone and wood) using one 1-MHz transducer. For thickness and velocity measurements, the WBP method gave a relative error lower than 1.5%. The relative errors in the mass density measurements ranged between 0.70% and 2.59%. Despite discrepancies between manufactured and biological samples, the results obtained on the three groups of samples using the WBP method in the reflection mode were remarkably consistent, indicating that it is a reliable and efficient means of simultaneously assessing the thickness and the velocity of the ultrasonic wave propagating in the medium, and the apparent mass density of material.

Analyzing the anisotropic Hooke's law for children's cortical bone.

Child cortical bone tissue is rarely studied because of the difficulty of obtaining samples. Yet the preparation and ultrasonic characterization of the small samples available, while challenging, is one of the most promising ways of obtaining information on the mechanical behavior of non-pathological children׳s bone. We investigated children׳s cortical bone obtained from chirurgical waste. 22 fibula or femur samples from 21 children (1-18 years old, mean age: 9.7±5.8 years old) were compared to 16 fibula samples from 16 elderly patients (50-95 years old, mean age: 76.2±13.5 years old). Stiffness coefficients were evaluated via an ultrasonic method and anisotropy ratios were calculated as the ratio of C33/C11, C33/C22 and C11/C22. Stiffness coefficients were highly correlated with age in children (R>0.56, p<0.01). No significant difference was found between C11 and C22 for either adult or child bone (p>0.5), nor between C44 and C55 (p>0.5). We observe a transverse isotropy with C33>C22=C11>C44C55>C66. For both groups, we found no correlation between age and anisotropy ratios. This study offers the first complete analysis of stiffness coefficients in the three orthogonal bone axes in children, giving some indication of how bone anisotropy is related to age. Future perspectives include studying the effect of the structure and composition of bone on its mechanical behavior.