Digital Image/Volume Correlation of Biological Tissues and Biomaterials

Fibrotic scarring is prevalent in a range of collagenous tissue disorders. Understanding the role of matrix biophysics in contributing to fibrotic progression is important to develop therapies, as well as to elucidate biological mechanisms. Here, we demonstrate how microfocus small-angle X-ray scattering (SAXS), with in situ mechanics and correlative imaging, can provide quantitative and position-resolved information on the fibrotic matrix nanostructure and its mechanical properties. We use as an example the case of keloid scarring in skin. SAXS mapping reveals heterogeneous gradients in collagen fibrillar concentration, fibril pre-strain (variations in D-period) and a new interfibrillar component likely linked to proteoglycans, indicating evidence of a complex 3D structure at the nanoscale. Furthermore, we demonstrate a proof-of-principle for a diffraction-contrast correlative imaging technique, incorporating, for the first time, DIC and SAXS, and providing an initial estimate for measuring spatially resolved fibrillar-level strain and reorientation in such heterogeneous tissues. By application of the method, we quantify (at the microscale) fibrillar reorientations, increases in fibrillar D-period variance, and increases in mean D-period under macroscopic tissue strains of ~20%. Our results open the opportunity of using synchrotron X-ray nanomechanical imaging as a quantitative tool to probe structure–function relations in keloid and other fibrotic disorders in situ. Full article

The mechanics of breathing is a fascinating and vital process. The lung has complexities and subtle heterogeneities in structure across length scales that influence mechanics and function. This study establishes an experimental pipeline for capturing alveolar deformations during a respiratory cycle using synchrotron radiation micro-computed tomography (SR-micro-CT). Rodent lungs were mechanically ventilated and imaged at various time points during the respiratory cycle. Pressure-Volume (P-V) characteristics were recorded to capture any changes in overall lung mechanical behaviour during the experiment. A sequence of tomograms was collected from the lungs within the intact thoracic cavity. Digital volume correlation (DVC) was used to compute the three-dimensional strain field at the alveolar level from the time sequence of reconstructed tomograms. Regional differences in ventilation were highlighted during the respiratory cycle, relating the local strains within the lung tissue to the global ventilation measurements. Strains locally reached approximately 150% compared to the averaged regional deformations of approximately 80–100%. Redistribution of air within the lungs was observed during cycling. Regions which were relatively poorly ventilated (low deformations compared to its neighbouring region) were deforming more uniformly at later stages of the experiment (consistent with its neighbouring region). Such heterogenous phenomena are common in everyday breathing. In pathological lungs, some of these non-uniformities in deformation behaviour can become exaggerated, leading to poor function or further damage. The technique presented can help characterize the multiscale biomechanical nature of a given pathology to improve patient management strategies, considering both the local and global lung mechanics. Full article

Strains within bone play an important role in the remodelling process and the mechanisms of fracture. The ability to assess these strains in vivo can provide clinically relevant information regarding bone health, injury risk, and can also be used to optimise treatments. In vivo bone strains have been investigated using multiple experimental techniques, but none have quantified 3D strains using non-invasive techniques. Digital volume correlation based on clinical MRI (DVC-MRI) is a non-invasive technique that has the potential to achieve this. However, before it can be implemented, uncertainties associated with the measurements must be quantified. Here, DVC-MRI was evaluated to assess its potential to measure in vivo strains in the talus. A zero-strain test (two repeated unloaded scans) was conducted using three MRI sequences, and three DVC approaches to quantify errors and to establish optimal settings. With optimal settings, strains could be measured with a precision of 200 με and accuracy of 480 με for a spatial resolution of 7.5 mm, and a precision of 133 με and accuracy of 251 με for a spatial resolution of 10 mm. These results demonstrate that this technique has the potential to measure relevant levels of in vivo bone strain and to be used for a range of clinical applications. Full article

Osteoarthritis (OA) is a chronic disease, affecting approximately one third of people over the age of 45. Whilst the etiology and pathogenesis of the disease are still not well understood, mechanics play an important role in both the initiation and progression of osteoarthritis. In this study, we demonstrate the application of stepwise compression, combined with microCT imaging and digital volume correlation (DVC) to measure and evaluate full-field strain distributions within osteoarthritic femoral heads under uniaxial compression. A comprehensive analysis showed that the microstructural features inherent in OA bone did not affect the level of uncertainties associated with the applied methods. The results illustrate the localization of strains at the loading surface as well as in areas of low bone volume fraction and subchondral cysts. Trabecular thickness and connectivity density were identified as the only microstructural parameters with any association to the magnitude of local strain measured at apparent yield strain or the volume of bone exceeding yield strain. This work demonstrates a novel approach to evaluating the mechanical properties of the whole human femoral head in case of severe OA. Full article

Hip implant failure is mainly due to aseptic loosening of the cotyle and is typically accompanied by defects in the acetabular region. Revision surgery aims to repair such defects before implantation by means of reconstruction materials, whose morselized bone graft represents the gold standard. Due to the limited availability of bone tissue, synthetic substitutes are also used. The aim of this study was to evaluate if a synthetic fully resorbable tri-calcium phosphate-based substitute can provide adequate mechanical stability when employed to restore severe, contained defects, in comparison with morselized bone graft. Five cadaveric pelvises were adopted, one side was reconstructed with morselized bone graft and the other with the synthetic substitute, consisting of dense calcium phosphate granules within a collagen matrix. During the biomechanical test, cyclic load packages of increasing magnitude were applied to each specimen until failure. Bone/implant motions were measured through Digital Image Correlation and were expressed in terms of permanent and inducible translations and rotations. The reconstruction types exhibited a similar behavior, consisting of an initial settling trend followed by failure as bone fracture (i.e., no failure of the reconstruction material). When 2.2 Body Weight was applied, the permanent translations were not significantly different between the two reconstructions (p = 0.06–1.0) and were below 1.0 mm. Similarly, the inducible translations did not differ significantly (p = 0.06–1.0) and were below 0.160 mm. Rotations presented the same order of magnitude but were qualitatively different. Overall, the synthetic substitute provided adequate mechanical stability in comparison with morselized bone graft, thus representing a reliable alternative to treat severe, contained acetabular defects. Full article

Cervical spine injuries (CSIs) arising from collisions are uncommon in contact sports, such as rugby union, but their consequences can be devastating. Several FE modelling approaches are available in the literature, but a fully calibrated and validated FE modelling framework for cervical spines under compressive dynamic-impact loading is still lacking and material properties are not adequately calibrated for such events. This study aimed to develop and validate a methodology for specimen-specific FE modelling of vertebral bodies under impact loading. Thirty-five (n = 35) individual vertebral bodies (VBs) were dissected from porcine spine segments, potted in bone cement and $\mu$CT scanned. A speckle pattern was applied to the anterior faces of the bones to allow digital image correlation (DIC), which monitored the surface displacements. Twenty-seven (n = 27) VBs were quasi-statically compressively tested to a load up to 10 $\mathrm{kN}$ from the cranial side. Specimen-specific FE models were developed for fourteen (n = 14) of the samples in this group. The material properties were optimised based on the experimental load-displacement data using a specimen-specific factor ($k_{G{S}_{static}}$) to calibrate a density to Young’s modulus relationship. The average calibration factor arising from this group was calculated (${\overline{K}}_{G{S}_{static}}$) and applied to a control group of thirteen (n = 13) samples. The resulting VB stiffnesses was compared to experimental findings. The final eight (n = 8) VBs were subjected to an impact load applied via a falling mass of $7.4$$\mathrm{k}\mathrm{g}$ at a velocity of $3.1$$\mathrm{m}$$\mathrm{s}$⁻¹. Surface displacements and strains were acquired from the anterior VB surface via DIC, and the impact load was monitored with two load cells. Specimen-specific FE models were created for this dynamic group and material properties were assigned again based on the density–Young’s modulus relationship previously validated for static experiments, supplemented with an additional factor ($K_{G{S}_{dynamic}}$). The optimised conversion factor for quasi-static loading, ${\overline{K}}_{G{S}_{static}}$, had an average of 0.033. Using this factor, the validation models presented an average numerical stiffness value $3.72$% greater than the experimental one. From the dynamic loading experiments, the value for $K_{G{S}_{dynamic}}$ was found to be 0.14, 4.2 times greater than ${\overline{K}}_{G{S}_{static}}$. The average numerical stiffness was $2.3$% greater than in the experiments. Almost all models presented similar stiffness variations and regions of maximum displacement to those observed via DIC. The developed FE modelling methodology allowed the creation of models which predicted both static and dynamic behaviour of VBs. Deformation patterns on the VB surfaces were acquired from the FE models and compared to DIC data, achieving high agreement. This methodology is now validated to be fully applied to create whole cervical spine models to simulate axial impact scenarios replicating rugby collision events. Full article

The survivorship of cementless orthopaedic implants may be related to their initial stability; insufficient press-fit can lead to excessive micromotion between the implant and bone, joint pain, and surgical revision. However, too much interference between implant and bone can produce excessive strains and damage the bone, which also compromises stability. An understanding of the nature and mechanisms of strain generation during implantation would therefore be valuable. Previous measurements of implantation strain have been limited to local discrete or surface measurements. In this work, we devise a Digital Volume Correlation (DVC) methodology to measure the implantation strain throughout the volume. A simplified implant model was implanted into analogue bone media using a customised loading rig, and a micro-CT protocol optimised to minimise artefacts due to the presence of the implant. The measured strains were interpreted by FE modelling of the displacement-controlled implantation, using a bilinear elastoplastic constitutive model for the analogue bone. The coefficient of friction between the implant and bone was determined using the experimental measurements of the reaction force. Large strains at the interface between the analogue bone and implant produced localised deterioration of the correlation coefficient, compromising the ability to measure strains in this region. Following correlation coefficient thresholding (removing strains with a coefficient less than 0.9), the observed strain patterns were similar between the DVC and FE. However, the magnitude of FE strains was approximately double those measured experimentally. This difference suggests the need for improvements in the interface failure model, for example, to account for localised buckling of the cellular analogue bone structure. A further recommendation from this work is that future DVC experiments involving similar geometries and structures should employ a subvolume size of 0.97 mm as a starting point. Full article

Personalised prosthetic sockets are fabricated by expert clinicians in a skill- and experience-based process, with research providing tools to support evidence-based practice. We propose that digital volume correlation (DVC) may offer a deeper understanding of load transfer from prosthetic sockets into the residual limb, and tissue injury risk. This study’s aim was to develop a transtibial amputated limb analogue for volumetric strain estimation using DVC, evaluating its ability to distinguish between socket designs. A soft tissue analogue material was developed, comprising silicone elastomer and sand particles as fiducial markers for image correlation. The material was cast to form an analogue residual limb informed by an MRI scan of a person with transtibial amputation, for whom two polymer check sockets were produced by an expert prosthetist. The model was micro-CT scanned according to (i) an unloaded noise study protocol and (ii) a case study comparison between the two socket designs, loaded to represent two-legged stance. The scans were reconstructed to give 108 µm voxels. The DVC noise study indicated a 64 vx subvolume and 50% overlap, giving better than 0.32% strain sensitivity, and ~3.5 mm spatial resolution of strain. Strain fields induced by the loaded sockets indicated tensile, compressive and shear strain magnitudes in the order of 10%, with a high signal:noise ratio enabling distinction between the two socket designs. DVC may not be applicable for socket design in the clinical setting, but does offer critical 3D strain information from which existing in vitro and in silico tools can be compared and validated to support the design and manufacture of prosthetic sockets, and enhance the biomechanical understanding of the load transfer between the limb and the prosthesis. Full article

Osteochondral injuries are increasingly prevalent, yet success in articular cartilage regeneration remains elusive, necessitating the development of new surgical interventions and novel medical devices. As part of device development, animal models are an important milestone in illustrating functionality of novel implants. Inspection of the tissue-biomaterial system is vital to understand and predict load-sharing capacity, fixation mechanics and micromotion, none of which are directly captured by traditional post-mortem techniques. This study aims to characterize the localised mechanics of an ex vivo ovine osteochondral tissue–biomaterial system extracted following six weeks in vivo testing, utilising laboratory micro-computed tomography, in situ loading and digital volume correlation. Herein, the full-field displacement and strain distributions were visualised across the interface of the system components, including newly formed tissue. The results from this exploratory study suggest that implant micromotion in respect to the surrounding tissue could be visualised in 3D across multiple loading steps. The methodology provides a non-destructive means to assess device performance holistically, informing device design to improve osteochondral regeneration strategies. Full article

Regenerative medicine solutions require thoughtful design to elicit the intended biological response. This includes the biomechanical stimulus to generate an appropriate strain in the scaffold and surrounding tissue to drive cell lineage to the desired tissue. To provide appropriate strain on a local level, new generations of scaffolds often involve anisotropic spatially graded mechanical properties that cannot be characterised with traditional materials testing equipment. Volumetric examination is possible with three-dimensional (3D) imaging, in situ loading and digital volume correlation (DVC). Micro-CT and DVC were utilised in this study on two sizes of 3D-printed inorganic/organic hybrid scaffolds (n = 2 and n = 4) with a repeating homogenous structure intended for cartilage regeneration. Deformation was observed with a spatial resolution of under 200 µm whilst maintaining displacement random errors of 0.97 µm, strain systematic errors of 0.17% and strain random errors of 0.031%. Digital image correlation (DIC) provided an analysis of the external surfaces whilst DVC enabled localised strain concentrations to be examined throughout the full 3D volume. Strain values derived using DVC correlated well against manually calculated ground-truth measurements (R² = 0.98, n = 8). The technique ensures the full 3D micro-mechanical environment experienced by cells is intimately considered, enabling future studies to further examine scaffold designs for regenerative medicine. Full article

Patients spend months between the primary spinal tumor diagnosis and the surgical treatment, due to the need for performing chemotherapy and/or radiotherapy. During this period, they are exposed to an unknown risk of fracture. The aim of this study was to assess if it is possible to measure the mechanical strain in vertebrae affected by primary tumors, so as to open the way to an evidence-based scoring or prediction tool. We performed biomechanical tests on three vertebrae with bone tumor removed from patients. The tests were designed so as not to compromise the standard surgical and diagnostic procedures. Non-destructive mechanical tests in combination with state-of-the-art digital image correlation allowed to measure the distribution of strain on the surface of the vertebra. Our study has shown that the strains in the tumor region is circa 3 times higher than in the healthy bones, with principal strain peaks of 40,000/−20,000 microstrain, indicating a stress concentration potentially triggering vertebral fracture. This study has proven it is possible to analyze the mechanical behavior of primary tumor vertebrae as part of the clinical treatment protocol. This will allow building a tool for quantifying the risk of fracture and improving decision making in spine tumors. Full article

The extracellular matrix (ECM) plays a key role during cell migration, proliferation, and differentiation by providing adhesion sites and serving as a physical scaffold. Elucidating the interaction between the cell and ECM can reveal the underlying mechanisms of cellular behavior that are currently unclear. Analysis of the deformation of the ECM due to cell–matrix interactions requires microscopic, three-dimensional (3-D) imaging methods, such as confocal microscopy and second-harmonic generation microscopy, which are currently limited by phototoxicity and bleaching as a result of the point-scanning approach. In this study, we suggest the use of optical coherence microscopy (OCM) as a live-cell, volumetric, fast imaging tool for analyzing the deformation of fibrous ECM. We optimized such OCM parameters as the sampling rate to obtain images of the best quality that meet the requirements for robust digital volume correlation (DVC) analysis. Visualization and analysis of the mechanical interaction between collagen ECM and human umbilical vein endothelial cells (HUVECs) show that cellular adhesion during protrusion can be analyzed and quantified. The advantages of OCM, such as fine isotropic spatial resolution, fast time resolution, and low phototoxicity, make it the ideal optic tool for 3-D traction force microscopy. Full article

A deeper understanding of the cartilage-bone mechanics is fundamental to unravel onset and progression of osteoarthritis, enabling better diagnosis and treatment. The aim of this study is therefore to explore the capability of X-ray computed (XCT) phase-contrast imaging in a lab-based system to enable digital volume correlation (DVC) measurements of unstained cartilage-bone plugs from healthy adult bovines. DVC strain uncertainties were computed for both articular cartilage and mineralized tissue (calcified cartilage and subchondral bone) in the specimens at increasing propagation distances, ranging from absorption up to four times (4× such effective distance. In addition, a process of dehydration and rehydration was proposed to improve feature recognition in XCT of articular cartilage and mechanical properties of this tissue during the process were assessed via micromechanical probing (indentation), which was also used to determine the effect of long X-ray exposure. Finally, full-field strain from DVC was computed to quantify residual strain distribution at the cartilage-bone interface following unconfined compression test (ex situ). It was found that enhanced gray-scale feature recognition at the cartilage-bone interface was achieved using phase-contrast, resulting in reduced DVC strain uncertainties compared to absorption. Residual strains up to ~7000 µε in the articular cartilage were transferred to subchondral bone via the calcified cartilage and micromechanics revealed the predominant effect of long phase-contrast X-ray exposure in reducing both stiffness and hardness of the articular cartilage. The results of this study will pave the way for further development and refinement of the techniques, improving XCT-based strain measurements in cartilage-bone and other soft-hard tissue interfaces. Full article

While the non-linear behavior of spine segments has been extensively investigated in the past, the behavior of the Anterior Longitudinal Ligament (ALL) and its contribution during flexion and extension has never been studied considering the spine as a whole. The aims of the present study were to exploit Digital Image Correlation (DIC) to: (I) characterize the strain distribution on the ALL during flexion-extension, (II) compare the strain on specific regions of interest (ROI) of the ALL in front of the vertebra and of the intervertebral disc, (III) analyze the non-linear relationship between the surface strain and the imposed rotation and the resultant moment. Three specimens consisting of 6 functional spinal units (FSUs) were tested in flexion-extension. The full-field strain maps were measured on the surface of the ALL, and the most strained areas were investigated in detail. The DIC-measured strains showed different values of peak strain in correspondence with the vertebra and the disc but the average over the ROIs was of the same order of magnitude. The strain-moment curves showed a non-linear response like the moment-angle curves: in flexion the slope of the strain-moment curve was greater than in extension and with a more abrupt change of slope. To the authors’ knowledge, this is the first study addressing, by means of a full-field strain measurement, the non-linear contribution of the ALL to spine biomechanics. This study was limited to only three specimens; hence the results must be taken with caution. This information could be used in the future to build more realistic numerical models of the spine. Full article