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Bone is a biological composite material consisting of two main components: collagen and mineral. Collagen is the most abundant protein in vertebrates, which makes it of high clinical and scientific interest. In this paper, we compare the composition and structure of cortical bone demineralized using several protocols: ethylene-diamine-tetraacetic acid (EDTA), formic acid (CH2O2), hydrochloric acid (HCl), and HCl/EDTA mixture. The efficiencies of these four agents were investigated by assessing the remaining mineral quantities and collagen integrity with various experimental techniques. Raman spectroscopy results show that the bone demineralized by the CH2O2 agent has highest collagen quality parameter. The HCl/EDTA mixture removes the most mineral, but it affects the collagen secondary structure as amide II bands are shifted as observed by Fourier transform infrared spectroscopy. Thermogravimetric analysis reveals that HCl and EDTA are most effective in removing the mineral with bulk measurements. In summary, we conclude that HCl best demineralizes bone, leaving the well-preserved collagen structure in the shortest time. These findings guide on the best demineralization protocol to obtain high-quality collagen from bone for clinical and scientific applications.

Bone is composed of mineral (apatite crystals) phase, organic (proteins, mainly collagen, and other organics) phase, and water at the nanoscale. Mineral contributes to elastic modulus and strength while organics and water give bone its toughness. Changes in the mineral composition and arrangement with age during development, and the resulting mechanical properties, are of high scientific and clinical interests. The mineral phase can be studied by removing the organic phase from bone using sodium hypochlorite (NaClO). This process is called deproteinization (DP). In this paper, four age groups (3-week, 4-week, 16-week, and 24-week) were compared in both untreated and DP bone states. We find that the deproteinized bone samples from all these age groups are self-standing. Thus, the mineral phase forms a continuous structure, even in the 3-week bone. Micro-computed tomography imaging displays a significant porosity decrease with the increase in age. Raman spectroscopy reveals increases in both the carbonate-to-phosphate ratio and the mineral crystallinity with age. Compression test results show that the elastic modulus and strength of the mineral phase alone are positively related to age. The elastic modulus of DP bone is less affected by age than that of the untreated bone. Overall, the mechanical properties of the mineral phase increase with age due to a decrease in porosity and an increase in the mineral content.

Multi-material 3D printing offers design flexibility for the manufacture of new architectured composite materials and has been increasingly used to explore the mechanical properties of bioinspired composites. In this study, the influence of scale and size on mechanical properties of periodic two-phase composites with various architectures of a stiff and a soft phase are explored. The studied composites include an interpenetrating phase composite with two continuous phases (inspired by bone), a matrix-inclusion composite with a continuous and a discontinuous phase, and a discontinuous phase composite where both phases are discontinuous. These composites are fabricated by additive manufacturing, and their mechanical properties are evaluated experimentally using a compression test complemented by digital image correlation, and numerically through finite element analysis. Overall trends show that the elastic moduli and yield strengths decrease as the scale is increased. Our results on size effects reveal an increase in elastic modulus and yield strength with increasing size of the sample. Our investigation outcomes help to better understand some of the challenges in studying and applications of additively manufactured composites and shed light on scale and size effects on mechanical properties of 3D printed two-phase composites. This paper illustrates a need for further studies of scale and size effects in additively manufactured materials.

Freeze casting under external fields (magnetic, electric, or acoustic) produces porous materials having local, regional, and global microstructural order in specific directions. In freeze casting, porosity is typically formed by the directional solidification of a liquid colloidal suspension. Adding external fields to the process allows for structured nucleation of ice and manipulation of particles during solidification. External control over the distribution of particles is governed by a competition of forces between constitutional supercooling and electromagnetism or acoustic radiation. Here, we review studies that apply external fields to create porous ceramics with different microstructural patterns, gradients, and anisotropic alignments. The resulting materials possess distinct gradient, core–shell, ring, helical, or long-range alignment and enhanced anisotropic mechanical properties.

Bone is a biological composite material having collagen and mineral as its main constituents. In order to better understand the arrangement of the mineral phase in bone, porcine cortical bone was deproteinized using different chemical treatments. This study aims to determine the best method to remove the protein constituent while preserving the mineral component. Chemicals used were H2O2, NaOCl, NaOH, and KOH, and the efficacy of deproteinization treatments was determined by thermogravimetric analysis and Raman spectroscopy. The structure of the residual mineral parts was examined using scanning electron microscopy. X-ray diffraction was used to confirm that the mineral component was not altered by the chemical treatments. NaOCl was found to be the most effective method for deproteinization and the mineral phase was self-standing, supporting the hypothesis that bone is an interpenetrating composite. Thermogravimetric analyses and Raman spectroscopy results showed the preservation of mineral crystallinity and presence of residual organic material after all chemical treatments. A defatting step, which has not previously been used in conjunction with deproteinization to isolate the mineral phase, was also used. Finally, Raman spectroscopy demonstrated that the inclusion of a defatting procedure resulted in the removal of some but not all residual protein in the bone.

We present an overview on additive manufacturing (AM), also called three-dimensional printing, with a focus on polymers. First, we introduce the AM concept. Next, we outline several AM processes, including their advantages and limitations, and list common polymers that are used in commercial printers. Then, we state various AM applications and present two examples. We conclude with a global view of the AM field, its challenges, and future directions.

Biological materials found in Nature such as nacre and bone are well recognized as light-weight, strong, and tough structural materials. The remarkable toughness and damage tolerance of such biological materials are conferred through hierarchical assembly of their multiscale (i.e., atomic- to macroscale) architectures and components. Herein, the toughening mechanisms of different organisms at multilength scales are identified and summarized: macromolecular deformation, chemical bond breakage, and biomineral crystal imperfections at the atomic scale; biopolymer fibril reconfiguration/deformation and biomineral nanoparticle/nanoplatelet/nanorod translation, and crack reorientation at the nanoscale; crack deflection and twisting by characteristic features such as tubules and lamellae at the microscale; and structure and morphology optimization at the macroscale. In addition, the actual loading conditions of the natural organisms are different, leading to energy dissipation occurring at different time scales. These toughening mechanisms are further illustrated by comparing the experimental results with computational modeling. Modeling methods at different length and time scales are reviewed. Examples of biomimetic designs that realize the multiscale toughening mechanisms in engineering materials are introduced. Indeed, there is still plenty of room mimicking the strong and tough biological designs at the multilength and time scale in Nature.

Freeze casting is a technique used to manufacture porous ceramics with aligned microstructures. In conventional freeze casting, these microstructures are aligned along a single direction of freezing. However, a caveat to these ceramics has been their ensuing lack of strength and toughness due to their high porosity, especially in the direction orthogonal to the direction of alignment. In this work, a novel freezing casting method referred to as &ldquo, radial-concentric freeze casting&rdquo, is presented, which takes its inspiration from the radially and concentrically aligned structure of the defensive spines of the porcupine fish. The method builds off the radial freeze casting method, in which the microstructure is aligned radially, and imposes a concentric alignment. Axial compression and Brazilian tests were performed to obtain axial compressive strengths, axial compressive moduli, and splitting tensile strengths of freeze cast samples with and without epoxy infiltration. Notably, radial-concentric freeze cast samples had the greatest improvements in axial compressive modulus and splitting tensile strength with infiltration, when compared against the changes in mechanical properties of conventional and radial freeze cast ceramics with infiltration. These results provide further evidence for the importance of structure in multiphase materials and the possibility of enhancing mechanical properties through the controlled alignment of microstructures.

Tarantulas extend the femur–patella (proximal) and tibia–metatarsal (distal) joints of their legs hydraulically. Because these two hydraulically actuated joints are positioned in series, hemolymph flow within each leg is expected to mechanically couple the movement of the joints. In the current study, we tested two hypotheses: (1) at lower temperatures, movement of the two in-series hydraulic joints within a leg will be less coupled because of increased hemolymph viscosity slowing hemolymph flow; and (2) at higher temperatures, movement of the two in-series hydraulic joints will be less coupled because the higher stride frequencies limit the time available for hemolymph flow. We elicited maximal running speeds at four ecologically relevant temperatures (15, 24, 31 and 40°C) in Texas Brown tarantulas (Aphonopelma hentzi). The spiders increased sprint speed 2.5-fold over the temperature range by changing their stride frequency but not stride length. The coefficient of determination for linear regression (R2) of the proximal and distal joint angles was used as the measure of the degree of coupling between the two joints. This coupling coefficient between the proximal and distal joint angles, for both forelegs and hind­legs, was significantly lowest at the highest temperature at which the animals ran the fastest with the highest stride frequencies. The coordination of multiple, in-series hydraulically actuated joints may be limited by operating speed.

This paper explores the structure, composition, and mechanical properties of porcupine fish spines for the first time. The spine was found to be composed of nanocrystalline hydroxyapatite, protein (collagen), and water using X-ray diffraction, energy-dispersive X-ray spectroscopy, and thermogravimetric analysis. Microstructures have mineralized fibrillar sheets in the longitudinal direction and in a radial orientation in the transverse direction that were observed using light and electron microscopy. Based on the images, the hierarchical structure of the spine shows both concentric and radial reinforcement. Mechanical properties were obtained using cantilever beam and nanoindentation tests. A tapered cantilever beam model was developed and compared to that of a uniform cantilever beam. The tapered beam model showed that while the stresses experienced were similar to those of the uniform beam, the location of the maximum stress was near the distal region of the beam rather than at the base, which allows the porcupine fish to conserve energy and resources if the spine is fractured.