Bioinspired rational design of bi-material 3D printed soft-hard interfaces.

Durable interfacing of hard and soft materials is a major design challenge caused by the ensuing stress concentrations. In nature, soft-hard interfaces exhibit remarkable mechanical performance, with failures rarely happening at the interface. Here, we mimic the strategies observed in nature to design efficient soft-hard interfaces. We base our geometrical designs on triply periodic minimal surfaces (i.e., Octo, Diamond, and Gyroid), collagen-like triple helices, and randomly distributed particles. A combination of computational simulations and experimental techniques, including uniaxial tensile and quad-lap shear tests, are used to characterize the mechanical performance of the interfaces. Our analyses suggest that smooth interdigitated connections, compliant gradient transitions, and either decreasing or constraining strain concentrations lead to simultaneously strong and tough interfaces. We generate additional interfaces where the abovementioned toughening mechanisms work synergistically to create soft-hard interfaces with strengths approaching the upper achievable limit and enhancing toughness values by 50%, as compared to the control group.

3D printed patient-specific fixation plates for the treatment of slipped capital femoral epiphysis: Topology optimization vs. conventional design.

Orthopedic plates are commonly used after osteotomies for temporary fixation of bones. Patient-specific plates have recently emerged as a promising fixation device. However, it is unclear how various strategies used for the design of such plates perform in comparison with each other. Here, we compare the biomechanical performance of 3D printed patient-specific bone plates designed using conventional computer-aided design (CAD) techniques with those designed with the help of topology optimization (TO) algorithms, focusing on cases involving slipped capital femoral epiphysis (SCFE). We established a biomechanical testing protocol to experimentally assess the performance of the designed plates while measuring the full-field strain using digital image correlation. We also created an experimentally validated finite element model to analyze the performance of the plates under physiologically relevant loading conditions. The results indicated that the TO construct exhibited higher ultimate load and biomechanical performance as compared to the CAD construct, suggesting that TO is a viable approach for the design of such patient-specific bone plates. The TO plate also distributed stress more evenly over the screws, likely resulting in more durable constructs and improved anatomical conformity while reducing the risk of screw and plate failure during cyclic loading. Although differences existed between finite element analysis and experimental testing, this study demonstrated that finite element modelling can be used as a reliable method for evaluating and optimizing plates for SCFE patients. In addition to enhancing the mechanical performance of patient-specific fixation plates, the utilization of TO in plate design may also improve the surgical outcome and decrease the recovery time by reducing the plate and incision sizes.

Biomechanical evaluation of additively manufactured patient-specific mandibular cage implants designed with a semi-automated workflow: A cadaveric and retrospective case study.

OBJECTIVE: Mandibular reconstruction using patient-specific cage implants is a promising alternative to the vascularized free flap reconstruction for nonirradiated patients with adequate soft tissues, or for patients whose clinical condition is not conducive to microsurgical reconstruction. This study aimed to assess the biomechanical performance of 3D printed patient-specific cage implants designed with a semi-automated workflow in a combined cadaveric and retrospective case series study. METHODS: We designed cage implants for two human cadaveric mandibles using our previously developed design workflow. The biomechanical performance of the implants was assessed with the finite element analysis (FEA) and quasi-static biomechanical testing. Digital image correlation (DIC) was used to measure the full-field strains and validate the FE models by comparing the distribution of maximum principal strains within the bone. The retrospective study of a case series involved three patients, each of whom was treated with a cage implant of similar design. The biomechanical performance of these implants was evaluated using the experimentally validated FEA under the scenarios of both mandibular union and nonunion. RESULTS: No implant or screw failure was observed prior to contralateral bone fracture during the quasi-static testing of both cadaveric mandibles. The FEA and DIC strain contour plots indicated a strong linear correlation (r = 0.92) and a low standard error (SE=29.32µÎµ), with computational models yielding higher strain values by a factor of 2.7. The overall stresses acting on the case series' implants stayed well below the yield strength of additively manufactured (AM) commercially pure titanium, when simulated under highly strenuous chewing conditions. Simulating a full union between the graft and remnant mandible yielded a substantial reduction (72.7±1.5%) in local peak stresses within the implants as compared to a non-bonded graft. CONCLUSIONS: This study shows the suitability of the developed semi-automated workflow in designing patient-specific cage implants with satisfactory mechanical functioning under demanding chewing conditions. The proposed workflow can aid clinical engineers in creating reconstruction systems and streamlining pre-surgical planning. Nevertheless, more research is still needed to evaluate the osteogenic potential of bone graft insertions.

Design considerations for patient-specific bone fixation plates: a literature review.

In orthopedic surgery, patient-specific bone plates are used for fixation when conventional bone plates do not fit the specific anatomy of a patient. However, plate failure can occur due to a lack of properly established design parameters that support optimal biomechanical properties of the plate.This review provides an overview of design parameters and biomechanical properties of patient-specific bone plates, which can assist in the design of the optimal plate.A literature search was conducted through PubMed and Embase, resulting in the inclusion of 78 studies, comprising clinical studies using patient-specific bone plates for fracture fixation or experimental studies that evaluated biomechanical properties or design parameters of bone plates. Biomechanical properties of the plates, including elastic stiffness, yield strength, tensile strength, and Poisson's ratio are influenced by various factors, such as material properties, geometry, interface distance, fixation mechanism, screw pattern, working length and manufacturing techniques.Although variations within studies challenge direct translation of experimental results into clinical practice, this review serves as a useful reference guide to determine which parameters must be carefully considered during the design and manufacturing process to achieve the desired biomechanical properties of a plate for fixation of a specific type of fracture.

Multi-scale in silico and ex silico mechanics of 3D printed cochlear implants for local drug delivery.

The currently available treatments for inner ear disorders often involve systemic drug administration, leading to suboptimal drug concentrations and side effects. Cochlear implants offer a potential solution by providing localized and sustained drug delivery to the cochlea. While the mechanical characterization of both the implants and their constituent material is crucial to ensure functional performance and structural integrity during implantation, this aspect has been mostly overlooked. This study proposes a novel methodology for the mechanical characterization of our recently developed cochlear implant design, namely, rectangular and cylindrical, fabricated using two-photon polymerization (2 PP) with a novel photosensitive resin (IP-Q™). We used in silico computational models and ex silico experiments to study the mechanics of our newly designed implants when subjected to torsion mimicking the foreseeable implantation procedure. Torsion testing on the actual-sized implants was not feasible due to their small size (0.6 × 0.6 × 2.4 mm³). Therefore, scaled-up rectangular cochlear implants (5 × 5 × 20 mm³, 10 × 10 × 40 mm³, and 20 × 20 × 80 mm³) were fabricated using stereolithography and subjected to torsion testing. Finite element analysis (FEA) accurately represented the linear behavior observed in the torsion experiments. We then used the validated Finite element analysis models to study the mechanical behavior of real-sized implants fabricated from the IP-Q resin. Mechanical characterization of both implant designs, with different inner porous structures (pore size: 20 µm and 60 µm) and a hollow version, revealed that the cylindrical implants exhibited approximately three times higher stiffness and mechanical strength as compared to the rectangular ones. The influence of the pore sizes on the mechanical behavior of these implant designs was found to be small. Based on these findings, the cylindrical design, regardless of the pore size, is recommended for further research and development efforts.

Bending analysis of sandwich panel composite with a re-entrant lattice core using zig-zag theory.

The sandwich panel structures have been widely used in many industrial applications because of their high mechanical properties. The middle layer of these structures is very important factor in controlling and enhancing their mechanical performance under various loading scenarios. The re-entrant lattice configurations, are prominent candidates that can be used as the middle layer in such sandwich structures because of several reasons namely the simplicity in tuning their elastic (e.g., values of Poisson's ratio and elastic stiffness) and plastic (e.g., high strength-to-weight ratio) properties by only adjusting the geometrical features of the constituting unit cells. Here, we investigated the response of a three-layered sandwich plate with a re-entrant core lattice under flexural bending using analytical (i.e., zig-zag theory), computational (i.e., finite element) and experimental tests. We also analyzed the effects of different geometrical parameters (e.g., angle, thicknesses, and length to the height ratio of unit cells) of re-entrant lattice structures on the overall mechanical behavior of sandwich structures. We found that the core structures with auxetic behavior (i.e., negative Poisson's ratio) resulted in a higher bending strength and a minimum out-of-plane shear stress as compared to those with conventional lattices. Our results can pave way in designing advanced engineered sandwich structures with architected core lattices for aerospace and biomedical applications.

Semi-automated digital workflow to design and evaluate patient-specific mandibular reconstruction implants.

The reconstruction of large mandibular defects with optimal aesthetic and functional outcomes remains a major challenge for maxillofacial surgeons. The aim of this study was to design patient-specific mandibular reconstruction implants through a semi-automated digital workflow and to assess the effects of topology optimization on the biomechanical performance of the designed implants. By using the proposed workflow, a fully porous implant (LA-implant) and a topology-optimized implant (TO-implant) both made of Ti-6Al-4V ELI were designed and additively manufactured using selective laser melting. The mechanical performance of the implants was predicted by performing finite element analysis (FEA) and was experimentally assessed by conducting quasi-static and cyclic biomechanical tests. Digital image correlation (DIC) was used to validate the FE model by comparing the principal strains predicted by the FEM model with the measured distribution of the same type of strain. The numerical predictions were in good agreement with the DIC measurements and the predicted locations of specimen failure matched the actual ones. No statistically significant differences (p < 0.05) in the mean stiffness, mean ultimate load, or mean ultimate displacement were detected between the LA- and TO-implant groups. No implant failures were observed during quasi-static or cyclic testing under masticatory loads that were substantially higher (>1000 N) than the average maximum biting force of healthy individuals. Given its relatively lower weight (16.5%), higher porosity (17.4%), and much shorter design time (633.3%), the LA-implant is preferred for clinical application. This study clearly demonstrates the capability of the proposed workflow to develop patient-specific implants with high precision and superior mechanical performance, which will greatly facilitate cost- and time-effective pre-surgical planning and is expected to improve the surgical outcome.

Mechanisms of fatigue crack initiation and propagation in auxetic meta-biomaterials.

The fatigue performance of additively manufactured auxetic meta-biomaterials made from commercially pure titanium has been studied only recently. While certain assumptions have been made regarding the mechanisms underlying their fatigue failure, the exact mechanisms are not researched yet. Here, we studied the mechanisms of crack formation and propagation in cyclically loaded auxetic meta-biomaterials. Twelve different designs were subjected to compression-compression fatigue testing while performing full-field strain measurement using digital image correlation (DIC). The fatigue tests were stopped at different points before complete specimen failure to study the evolution of damage in the micro-architecture of the specimens using micro-computed tomography (micro-CT). Furthermore, finite element models were made to study the presence of stress concentrations. Structural weak spots were found in the inverted nodes and the vertical struts located along the outer rim of the specimens, matching the maximum principal strain concentrations and fracture sites in the DIC and micro-CT data. Cracks were often found to originate from internal void spaces or from sites susceptible to mode-I cracking. Many specimens maintained their structural integrity and exhibited no signs of rapid strain accumulation despite the presence of substantial crack growth. This observation underlines the importance of such microscale studies to identify accumulated damage that otherwise goes unnoticed. The potential release of powder particles from damaged lattices could elicit a foreign body response, adversely affecting the implant success. Finding the right failure criterion, therefore, requires more data than only those pertaining to macroscopic measurements and should always include damage assessment at the microscale. STATEMENT OF SIGNIFICANCE: The negative Poisson's ratio of auxetic meta-biomaterials makes them expand laterally in response to axial tension. This extraordinary property has great potential in the field of orthopedics, where it could enhance bone-implant contact. The fatigue performance of additively manufactured auxetic meta-biomaterials has only recently been studied and was found to be superior to many other bending- and stretch-dominated micro-architectures. In this study, we go beyond these macroscopic measurements and focus on the crack initiation and propagation. Full-field strain measurements and 3D imaging are used to paint a detailed picture of the mechanisms underlying fatigue. Using these data, specific aspects of the design and/or printing process can be targeted to improve the performance of auxetic meta-biomaterials in load-bearing applications.

3D printed submicron patterns orchestrate the response of macrophages.

The surface topography of engineered extracellular matrices is one of the most important physical cues regulating the phenotypic polarization of macrophages. However, not much is known about the ways through which submicron (i.e., 100-1000 nm) topographies modulate the polarization of macrophages. In the context of bone tissue regeneration, it is well established that this range of topographies stimulates the osteogenic differentiation of stem cells. Since the immune response affects the bone tissue regeneration process, the immunomodulatory consequences of submicron patterns should be studied prior to their clinical application. Here, we 3D printed submicron pillars (using two-photon polymerization technique) with different heights and interspacings to perform the first ever systematic study of such effects. Among the studied patterns, the highest degree of elongation was observed for the cells cultured on those with the tallest and densest pillars. After 3 days of culture with inflammatory stimuli (LPS/IFN-γ), sparsely decorated surfaces inhibited the expression of the pro-inflammatory cellular marker CCR7 as compared to day 1 and to the other patterns. Furthermore, sufficiently tall pillars polarized the M1 macrophages towards a pro-healing (M2) phenotype, as suggested by the expression of CD206 within the first 3 days. As some of the studied patterns are known to be osteogenic, the osteoimmunomodulatory capacity of the patterns should be further studied to optimize their bone tissue regeneration performance.

Comparison in clinical performance of surgical guides for mandibular surgery and temporomandibular joint implants fabricated by additive manufacturing techniques.

Additive manufacturing (AM) offers great design freedom that enables objects with desired unique and complex geometry and topology to be readily and cost-effectively fabricated. The overall benefits of AM are well known, such as increased material and resource efficiency, enhanced design and production flexibility, the ability to create porous structures and on-demand manufacturing. When AM is applied to medical devices, these benefits are naturally assumed. However, hard clinical evidence collected from clinical trials and studies seems to be lacking and, as a result, systematic assessment is yet difficult. In the present work, we have reviewed 23 studies on the clinical use of AM patient-specific surgical guides (PSGs) for the mandible surgeries (n = 17) and temporomandibular joint (TMJ) patient-specific implants (PSIs) (n = 6) with respect to expected clinical outcomes. It is concluded that the data published on these AM medical devices are often lacking in comprehensive evaluation of clinical outcomes. A complete set of clinical data, including those on time management, costs, clinical outcomes, range of motion, accuracy of the placement with respect to the pre-operative planning, and extra complications, as well as manufacturing data are needed to demonstrate the real benefits gained from applying AM to these medical devices and to satisfy regulatory requirements.

Fatigue performance of auxetic meta-biomaterials.

Meta-biomaterials offer a promising route towards the development of life-lasting implants. The concept aims to achieve solutions that are ordinarily impossible, by offering a unique combination of mechanical, mass transport, and biological properties through the optimization of their small-scale geometrical and topological designs. In this study, we primarily focus on auxetic meta-biomaterials that have the extraordinary ability to expand in response to axial tension. This could potentially improve the longstanding problem of implant loosening, if their performance can be guaranteed in cyclically loaded conditions. The high-cycle fatigue performance of additively manufactured (AM) auxetic meta-biomaterials made from commercially pure titanium (CP-Ti) was therefore studied. Small variations in the geometry of the re-entrant hexagonal honeycomb unit cell and its relative density resulted in twelve different designs (relative density: ~5-45%, re-entrant angle = 10-25°, Poisson's ratio = -0.076 to -0.504). Micro-computed tomography, scanning electron microscopy and mechanical testing were used to respectively measure the morphological and quasi-static properties of the specimens before proceeding with compression-compression fatigue testing. These auxetic meta-biomaterials exhibited morphological and mechanical properties that are deemed appropriate for bone implant applications (elastic modulus = 66.3-5648 MPa, yield strength = 1.4-46.7 MPa, pore size = 1.3-2.7 mm). With an average maximum stress level of 0.47 σy at 106 cycles (range: 0.35 σyσy- 0.82 σyσy), the auxetic structures characterized here are superior to many other non-auxetic meta-biomaterials made from the same material. The optimization of the printing process and the potential application of post-processing treatments could improve their performance in cyclically loaded settings even further. STATEMENT OF SIGNIFICANCE: Auxetic meta-biomaterials have a negative Poisson's ratio and, therefore, expand laterally in response to axial tension. Recently, they have been found to restore bone-implant contact along the lateral side of a hip stem. As a result, the bone will be compressed along both of the implant's contact lines, thereby actively reducing the risk of implant failure. In this case the material will be subjected to cyclic loading, for which no experimental data has been reported yet. Here, we present the first ever study of the fatigue performance of additively manufactured auxetic meta-biomaterials based on the re-entrant hexagonal honeycomb. These results will advance the adoption of auxetic meta-biomaterials in load-bearing applications, such as the hip stem, to potentially improve implant longevity.

Non-affinity in multi-material mechanical metamaterials.

Non-affine deformations enable mechanical metamaterials to achieve their unusual properties while imposing implications for their structural integrity. The presence of multiple phases with different mechanical properties results in additional non-affinity of the deformations, a phenomenon that has never been studied before in the area of extremal mechanical metamaterials. Here, we studied the degree of non-affinity, [Formula: see text], resulting from the random substitution of a fraction of the struts,[Formula: see text], that make up a lattice structure and are printed using a soft material (elastic modulus = [Formula: see text]) by those printed using a hard material ([Formula: see text]). Depending on the unit cell angle (i.e., [Formula: see text] = 60°, 90°, or 120°), the lattice structures exhibited negative, near-zero, or positive values of the Poisson's ratio, respectively. We found that the auxetic structures exhibit the highest levels of non-affinity, followed by the structures with positive and near-zero values of the Poisson's ratio. We also observed an increase in [Formula: see text] with [Formula: see text] and [Formula: see text] until [Formula: see text] =104 and [Formula: see text]= 75%-90% after which [Formula: see text] saturated. The dependency of [Formula: see text] upon [Formula: see text] was therefore found to be highly asymmetric. The positive and negative values of the Poisson's ratio were strongly correlated with [Formula: see text]. Interestingly, achieving extremely high or extremely low values of the Poisson's ratio required highly affine deformations. In conclusion, our results clearly show the importance of considering non-affinity when trying to achieve a specific set of mechanical properties and underscore the structural integrity implications in multi-material mechanical metamaterials.

Multi-material additive manufacturing technologies for Ti-, Mg-, and Fe-based biomaterials for bone substitution.

The growing interest in multi-functional metallic biomaterials for bone substitutes challenges the current additive manufacturing (AM, =3D printing) technologies. It is foreseeable that advances in multi-material AM for metallic biomaterials will not only allow for complex geometrical designs, but also improve their multi-functionalities by tuning the types or compositions of the underlying base materials, thereby presenting unprecedented opportunities for advanced orthopedic treatments. AM technologies are yet to be extensively explored for the fabrication of multi-functional metallic biomaterials, especially for bone substitutes. The aim of this review is to present the viable options of the state-of-the-art multi-material AM for Ti-, Mg-, and Fe-based biomaterials to be used as bone substitutes. The review starts with a brief review of bone tissue engineering, the design requirements, and fabrication technologies for metallic biomaterials to highlight the advantages of using AM over conventional fabrication methods. Five AM technologies suitable for metal 3D printing are compared against the requirements for multi-material AM. Of these AM technologies, extrusion-based multi-material AM is shown to have the greatest potential to meet the requirements for the fabrication of multi-functional metallic biomaterials. Finally, recent progress in the fabrication of Ti-, Mg-, and Fe-based biomaterials including the utilization of multi-material AM technologies is reviewed so as to identify the knowledge gaps and propose the directions of further research for the development of multi-material AM technologies that are applicable for the fabrication of multi-functional metallic biomaterials. STATEMENT OF SIGNIFICANCE: Addressing a critical bone defect requires the assistance of multi-functional porous metallic bone substitutes. As one of the most advanced fabrication technology in bone tissue engineering, additive manufacturing is challenged for its viability in multi-material fabrication of metallic biomaterials. This article reviews how the current metal additive manufacturing technologies have been and can be used for multi-material fabrication of Ti-, Mg-, and Fe-based bone substitutes. Progress on the Ti-, Mg-, and Fe-based biomaterials, including the utilization of multi-material additive manufacturing, are discussed to direct future research for advancing the multi-functional additively manufactured metallic bone biomaterials.

3D Printing of Large Areas of Highly Ordered Submicron Patterns for Modulating Cell Behavior.

Fabricating large areas of geometrically complex and precisely controlled topographies is required for the studies of cell behavior on patterned surfaces. Direct laser writing (DLW) is an advanced 3D-fabrication technique, which facilitates the manufacturing of structures within various scales (from a few hundred nanometers to millimeters). However, this method requires improvements in the accuracy and reproducibility of the submicron and nanoscale features that are printed over a large area. Here, we present a scheme to both improve the uniformity of the printed submicron patterns and decrease the printing time. The effects of various processing parameters (e.g., laser power and writing field) on the dimensions and uniformity of submicron pillars as well as on their Young's modulus and surface wettability were assessed. Decreasing the writing field to 33 × 33 µm2 significantly improved the uniformity of submicron pillars that were printed over an area of 4 mm2 in a single-step process. Preosteoblast cells (MC3T3-E1) were used to assess the cytocompatibility of the used material (IP-L780 resin) with a focus on cell morphology, cell proliferation, cytoskeletal organization, and the elastic modulus of the cells. The cells cultured for 2 days on the submicron pillars showed a polarized shape and a higher Young's modulus of the area corresponding to the nucleus relative to those cultured on flat surfaces. Taken together, the results of the current study clearly show that the submicron patterns created using DLW are both cytocompatible and could modulate the morphology and mechanical properties of cells. This work paves the way for direct printing of submicron features with controlled Young's moduli over large areas in a single-step process, which is necessary for systematically studying how such patterns modulate cellular functions.

Length-scale dependency of biomimetic hard-soft composites.

Biomimetic composites are usually made by combining hard and soft phases using, for example, multi-material additive manufacturing (AM). Like other fabrication methods, AM techniques are limited by the resolution of the device, hence, setting a minimum length scale. The effects of this length scale on the performance of hard-soft composites are not well understood. Here, we studied how this length scale affects the fracture toughness behavior of single-edge notched specimens made using random, semi-random, and ordered arrangements of the hard and soft phases with five different ratios of hard to soft phases. Increase in the length scale (40 to 960 µm) was found to cause a four-fold drop in the fracture toughness. The effects of the length scale were also modulated by the arrangement and volumetric ratio of both phases. A decreased size of the crack tip plastic zone, a crack path going through the soft phase, and highly strained areas far from the crack tip were the main mechanisms explaining the drop of the fracture toughness with the length scale.

In-silico quest for bactericidal but non-cytotoxic nanopatterns.

Nanopillar arrays that are bactericidal but not cytotoxic against the host cells could be used in implantable medical devices to prevent implant-associated infections. It is, however, unclear what heights, widths, interspacing, and shape should be used for the nanopillars to achieve the desired antibacterial effects while not hampering the integration of the device in the body. Here, we present an in-silico approach based on finite element modeling of the interactions between Staphylococcus aureus and nanopatterns on the one hand and osteoblasts and nanopatterns on the other hand to find the best design parameters. We found that while the height of the nanopillars seems to have little impact on the bactericidal behavior, shorter widths and larger interspacings substantially increase the bactericidal effects. The same combination of parameters could, however, also cause cytotoxicity. Our results suggest that a specific combination of height (120 nm), width (50 nm), and interspacing (300 nm) offers the bactericidal effects without cytotoxicity.

Shape-matching soft mechanical metamaterials.

Architectured materials with rationally designed geometries could be used to create mechanical metamaterials with unprecedented or rare properties and functionalities. Here, we introduce "shape-matching" metamaterials where the geometry of cellular structures comprising auxetic and conventional unit cells is designed so as to achieve a pre-defined shape upon deformation. We used computational models to forward-map the space of planar shapes to the space of geometrical designs. The validity of the underlying computational models was first demonstrated by comparing their predictions with experimental observations on specimens fabricated with indirect additive manufacturing. The forward-maps were then used to devise the geometry of cellular structures that approximate the arbitrary shapes described by random Fourier's series. Finally, we show that the presented metamaterials could match the contours of three real objects including a scapula model, a pumpkin, and a Delft Blue pottery piece. Shape-matching materials have potential applications in soft robotics and wearable (medical) devices.

Crumpling-based soft metamaterials: the effects of sheet pore size and porosity.

Crumpled-based materials are relatively easy to fabricate and show robust mechanical properties for practical applications, including meta-biomaterials design aimed for improved tissue regeneration. For such requests, however, the structure needs to be porous. We introduce a crumpled holey thin sheet as a robust bio-metamaterial and measure the mechanical response of a crumpled holey thin Mylar sheet as a function of the hole size and hole area fraction. We also study the formation of patterns of crease lines and ridges. The area fraction largely dominated the crumpling mechanism. We also show, the crumpling exponents slightly increases with increasing the hole area fraction and the total perimeter of the holes. Finally, hole edges were found to limit and guide the propagation of crease lines and ridges.

Characterizing microcrack orientation distribution functions in osteonal bone samples.

Prefailure microdamage in bone tissue is considered to be the most detrimental factor in defining its strength and toughness with respect to age and disease. To understand the influence of microcracks on bone mechanics it is necessary to assess their morphology and three-dimensional distribution. This requirement reaches beyond classic histology and stereology, and methods to obtain such information are currently missing. Therefore, the aim of the study was to develop a methodology that allows to characterize three-dimensional microcrack distributions in bulk bone samples. Four dumbbell-shaped specimens of human cortical bone of a 77-year-old female donor were loaded beyond yield in either tension, compression or torsion (one control). Subsequently, synchrotron radiation micro-computed tomography (SRµCT) was used to obtain phase-contrast images of the damaged samples. A microcrack segmentation algorithm was developed and used to segment microcrack families for which microcrack orientation distribution functions were determined. Distinct microcrack families were observed for each load case that resulted in distinct orientation distribution functions. Microcracks had median areas of approximately 4.7 µm2 , 33.3 µm2 and 64.0 µm2 for tension, compression and torsion. Verifying the segmentation algorithm against a manually segmented ground truth showed good results when comparing the microcrack orientation distribution functions. A size dependence was noted when investigating the orientation distribution functions with respect to the size of the volume of interest used for their determination. Furthermore, a scale separation between tensile, compressive and torsional microcracks was noticeable. Visual comparison to classic histology indicated that microcrack families were successfully distinguished. We propose a methodology to analyse three-dimensional microcrack distributions in overloaded cortical bone. Such information could improve our understanding of bone microdamage and its impact on bone failure in relation to tissue age and disease.