Bone metastases radiographically appear as regions with high (i.e. blastic metastases) or low (i.e. lytic metastases) bone mineral density. The clinical assessment of metastatic features is based on computed tomography (CT) but it is still unclear if the actual size of the metastases can be accurately detected from the CT images and if the microstructure in regions surrounding the metastases is altered (Nägele et al., 2004, Calc Tiss Int). This study aims to evaluate (i) the capability of the CT in evaluating the metastases size and (ii) if metastases affect the bone microstructure around them.

Ten spine segments consisted of a vertebra with lytic or mixed metastases and an adjacent control (radiologically healthy) were obtained through an ethically approved donation program. The specimens were scanned with a clinical CT (AquilionOne, Toshiba: slice thickness:1mm, in-plane resolution:0.45mm) to assess clinical metastatic features and a micro-CT (VivaCT80, Scanco, isotropic voxel size:0.039mm) to evaluate the detailed microstructure. The volume of the metastases was measured from both CT and micro-CT images (Palanca et al., 2021, Bone) and compared with a linear regression. The microstructural alteration around the metastases was evaluated in the volume of interest (VOI) defined in the micro-CT images as the volume of the vertebral body excluding the metastases. Three 3D microstructural parameters were calculated in the VOI (CTAn, Bruker SkyScan): Bone Volume Fraction (BV/TV), Trabecular Thickness (Tb.Th.), Trabecular Spacing (Tb.Sp.). Medians of each parameter were compared (Kruskal-Wallis, p=0.05).

One specimen was excluded as it was not possible to define the size of the metastases in the CT scans. A strong correlation between the volume obtained from the CT and micro-CT images was found (R2=0.91, Slope=0.97, Intercept=2.55, RMSE=5.7%, MaxError=13.12%). The differences in BV/TV, Tb.Th. and Tb.Sp. among vertebrae with lytic and mixed metastases and control vertebrae were not statistically significant (p-value>0.6). Similar median values of BV/TV were found in vertebrae with lytic (13.2±2.4%) and mixed (12.8±9.8%) metastases, and in controls (13.0±10.1%). The median Tb.Th. was 176±18 ∓m, 179±43 ∓m and 167±91 ∓m in vertebrae with lytic and mixed metastases and control vertebrae, respectively. The median Tb. Sp. was 846±26 ∓m, 849±286 ∓m and 880±116 ∓m in vertebrae with lytic and mixed metastases and control vertebrae, respectively.

In conclusion, the size of vertebral metastases can be accurately assess using CT images. The 3D microstructural parameters measured were comparable with those reported in the literature for healthy vertebrae (Nägele et al., 2004, Calc Tiss Int, Sone et al., 2004, Bone) and showed how the microstructure of the bone tissue surrounding the lesion is not altered by the metastases.

DVC allowed measurements of displacement and strain distribution in bone through the comparison of two, or more, 3D images. Hence, it has a potential as a diagnostic tool in combination with clinical CT. Currently, traditional computed tomography (CT) allows for a detailed 3D analysis of hard tissues, but imaging in a weight-bearing condition is still limited. PedCAT-CT (Curvebeam, USA) emerged as a novel technology allowing, for the first time, 3D imaging under full-weight bearing (Richter, Zech et al. 2015). Specifically, a PedCAT-CT based DVC was employed to establish its reliability through the strain uncertainties produced on bone structure targets, preliminarily to any further clinical studies. In addition, a reverse engineering FE modeling was used to predict possible force associated to displacement errors from DVC.

Three porcine thoracic vertebrae were used as bone benchmark for the DVC (Palanca, Tozzi et al. 2016, Tozzi, Dall'Ara et al. 2016). The choice of using porcine vertebrae (in a CT designed for foot/ankle) was driven by availability, as well as similar dimensions to the calcaneus. Each vertebra was immersed in saline solution and scanned twice without any repositioning (zero-strain-test) with a pedCAT-CT (Curvebeam, USA) obtaining an isotropic voxel size of 370 micrometers. Volumes of interest of 35 voxel were cropped inside the vertebrae. Displacement and strains were evaluated using DVC (DaVis-DC, LaVision, Germany), with different spatial resolution. The displacement maps were used to predict the force uncertainties via FE (Ansys Mechanical v.14, Ansys Inc, Canonsburg, PA). Each element was assigned a linear elastic isotropic constitutive law (Young modulus: 8 GPa, Poisson's ratio: 0.3, as in (Follet, Peyrin et al. 2007)). Overall, the precision error of strain measurement was evaluated as the average of the standard deviation of the absolute value of the different component of strain (Liu and Morgan 2007).

The force uncertainties obtained with the FE analysis produced magnitudes ranging from 231 to 2376 N. No clear trend on the force was observed in relation to the spatial resolution. Precision errors were smaller than 1000 microstrain in all cases, with the lowest ranging from 83 microstrain for the largest spatial resolution. Full-field strain on the bone tissue did not seem to highlight a preferential distribution of error in the volume.

The precision errors showed that the pedCAT-CT based DVC can be sufficient to investigate the bone tissue failure (7000–10000 microstrain) or, physiological deformation if well-optimized. FE analysis produced important force uncertainties up to 2376 N. However, this is a preliminary investigation. Further investigation will give a clearer indication on DVC based PedCAT-CT, as well as force uncertainties predicted. So far, the DVC showed its ability to measure displacement and strain with reasonable reliability with clinical-CT as well.

DVC is a novel full-field and contactless measurement technique for calculating displacements and strains inside bones (Grassi and Isaksson 2015) through the comparison of 3D reconstructions (CT, micro-CT, MRI, etc.) from unloaded and loaded samples. Recent in zero-strain tests to estimate the measurement precision by applying a known state of strain (Palanca, Tozzi et al. 2015) suggested that DVC is suitable to identify regions where bone tissue is yielded (i.e. subjected to high strains). Conversely to reliably measure strain in the physiological range a severe compromise with spatial resolution is necessary (Dall'Ara, Barber et al. 2014, Palanca, Tozzi et al. 2015). In order to use DVC to explore the relationship between the local physiological strain and bone microarchitecture, an error lower than 200 microstrain (an order of magnitude lower than the mean strain) and a spatial resolution of the strain measurement lower than 100 μm is required. The aim of this work is to define if, and to what extend, high-quality images obtained by synchrotron radiation micro computed tomography (SR-μCT) improve the precision of a global DVC approach.

Cylindrical specimens of cortical and trabecular bone were extracted from a fresh bovine femur and embedded in acrylic resin. Both samples were scanned twice without any repositioning (‘repeated scantest’) at beamline l13–2 of Diamond Light Source (Oxford, UK). 4000 projections of 53 ms exposure were collected via fly-scanning with a CdWO₄scintillator-coupled pco.edge 5.5 detector with 4× magnification and an effective pixel size of 1.6μm. Strains were evaluated using a global DVC approach (ShIRT-FE) in two cubic volumes of interest (VOI) of 1,000 voxels in side length, for each specimen, exploring a DVC spatial resolution from 16 to 498 μm. The precision of measurements was evaluated extracting a similar indicator to (Liu and Morgan 2007).

Precision improved with decreasing spatial resolution, confirming a trend similar to that obtained with ‘laboratory source’ μCT on similar specimens (Palanca, Tozzi et al. 2015). To obtain a precision of better than 200 microstrains the cortical and trabecular samples required spatial resolutions of 41 and 80 μm respectively. Comparing these results to those of previous studies, where similar specimens were scanned with ‘laboratory source’ μCT (effective voxel size of the order of ten μm) the errors were vastly reduced (approximately one order of magnitude). In fact, in order to obtain a precision of better than 200 microstrain, spatial resolutions of 550 (cortical) and 480 (trabecular) μm were needed (Dall'Ara, Barber et al. 2014).

This work showed that using high-quality tomograms obtained by synchrotron radiation μCT decreases the measurement uncertainties of a global DVC approach with respect to those obtained with laboratory source μCT. DVC could therefore be used with μCT data to evaluate displacement and strain in the physiological range with remarkable spatial resolution.

The causes of spine disease are often biomechanical ones (e.g. disc degeneration, vertebral fracture). Currently, a deep investigation of the spine biomechanics is missing, due to the high complexity of the spine system (Fung 1980, Brandolini, Cristofolini et al. 2014): vertebrae and intervertebral discs. Recently, the Digital Image Correlation allowed measuring in vitrothe displacement and strain on the surface of soft and hard tissues, upon a specific non-invasive preparation of their surface with a speckle pattern (Palanca, Tozzi et al. 2016). The aim of this explorative work was to evaluate the deformation on spine segments, being able to distinguish between hard and soft tissue in the elastic regime and up to fracture.

Segment of four vertebrae were extracted from porcine spines. All ligaments and muscles were removed, without damaging the spine segment (vertebrae and intervertebral discs). A suitable non-conventional white-on-black speckle pattern was prepared on the surface with airbrush airgun to track the movements of the specimen with DIC (Lionello, Sirieix et al. 2014). The endplates of the extreme vertebrae were potted in poly-methyl-methacrylate. The spine segments were tested in pure axial loading with cycles of increasing magnitude, up to fialure. A commercial 3D-DIC (Dantec Dynamics, Denmark) was used. In the present configuration, it allowed a resolution of 30 micrometers. It was used to measure the displacements and strains in a full-field and contactless way on the frontal surface of the spine segments.

DIC allowed measuring with success the displacement and strain during the entire test, in the elastic regime and up to failure. The displacements and strains could be measured on the entire specimen, both in the vertebrae (hard tissue) and in the intervertebral discs (soft tissue). The axial strain evaluated prior to failure was close to 10’000 microstrain on the vertebral body surface and exceed 70’000 microstrain on the intervertebral discs, where failure was localized.

The pattern, prepared in a dedicated way showed its suitability for both the bone and the disc. The evaluated failure strains were in agreement with the literature (Bayraktar, Morgan et al. 2004) (Spera, Genovese et al. 2011). To the authors' best knowledge, this kind of measurement including strain on soft and hard tissue simultaneously has never been performed before. This work showed the capability of DIC in providing full-field measures on the surface with complex geometry, such as the spine. The assertion of these potentialities could open the way to further application of DIC to study the behaviour of human spines, including improvement of spinal fixation devices.