Metal additive manufacturing is a powerful tool for providing the desired functional performance through a three-dimensional (3D) structural design. Among the material functions, anisotropic mechanical properties are indispensable for enabling the capabilities of structural materials for living tissues. For biomedical materials to replace bone function, it is necessary to provide an anisotropic mechanical property that mimics that of bones. For desired control of the mechanical performance of the materials, we propose a novel 3D puzzle structure with cube-shaped parts comprising 27 (3 × 3 × 3) unit compartments. We designed and fabricated a Co–Cr–Mo composite structure through spatial control of the positional arrangement of powder/solid parts using the laser powder bed fusion (L-PBF) method. The mechanical function of the fabricated structure can be predicted using the rule of mixtures based on the arrangement pattern of each part. The solid parts in the cubic structure were obtained by melting and solidifying the metal powder with a laser, while the powder parts were obtained through the remaining nonmelted powders inside the structure. This is the first report to achieve an innovative material design that can provide an anisotropic Young’s modulus by arranging the powder and solid parts using additive manufacturing technology.

In this study, a Ti–6Al–4V alloy composite with uniaxial anisotropy and a hierarchical structure is fabricated using electron beam powder bed fusion, one of the additive manufacturing techniques that enable arbitrary fabrication, and subsequent heat treatment. The uniaxial anisotropic deformation behavior and mechanical properties such as Young’s modulus are obtained by introducing a unidirectional honeycomb structure. The main feature of this structure is that the unmelted powder retained in the pores of the honeycomb structure. After appropriate heat treatment at 1020 °C, necks are formed between the powder particles and between the powder particles and the honeycomb wall, enabling a stress transmission through the necks when the composite is loaded. This means that the powder part has been mechanically functionalized by the neck formation. As a result, a plateau region appears in the stress–strain curve. The stress transfer among the powder particles leads to the cooperative deformation of the composites, contributing to the excellent energy absorption capacity. Therefore, it is expected that the composite can be applied to bone plates on uniaxially oriented microstructures such as long bones owing to its excellent energy absorption capacity and low elasticity to unidirectionally suppress stress shielding.

Human-induced pluripotent stem cells (hiPSCs) can be applied in patient-specific cell therapy to regenerate lost tissue or organ function. Anisotropic control of the structural organization in the newly generated bone matrix is pivotal for functional reconstruction during bone tissue regeneration. Recently, we revealed that hiPSC-derived osteoblasts (hiPSC-Obs) exhibit preferential alignment and organize in highly ordered bone matrices along a bone-mimetic collagen scaffold, indicating their critical role in regulating the unidirectional cellular arrangement, as well as the structural organization of regenerated bone tissue. However, it remains unclear how hiPSCs exhibit the cell properties required for oriented tissue construction. The present study aimed to characterize the properties of hiPSCs-Obs and those of their focal adhesions (FAs), which mediate the structural relationship between cells and the matrix. Our in vitro anisotropic cell culture system revealed the superior adhesion behavior of hiPSC-Obs, which exhibited accelerated cell proliferation and better cell alignment along the collagen axis compared to normal human osteoblasts. Notably, the oriented collagen scaffold stimulated FA formation along the scaffold collagen orientation. This is the first report of the superior cell adhesion behavior of hiPSC-Obs associated with the promotion of FA assembly along an anisotropic scaffold. These findings suggest a promising role for hiPSCs in enabling anisotropic bone microstructural regeneration.

Functional adaptation refers to the active modification of bone structure according to the mechanical loads applied daily to maintain its mechanical integrity and adapt to the environment. Functional adaptation relates to bone mass, bone mineral density (BMD), and bone morphology (e.g., trabecular bone architecture). In this study, we discovered for the first time that another form of bone functional adaptation of a cortical bone involves a change in bone quality determined by the preferential orientation of apatite nano-crystallite, a key component of the bone. An in vivo rat ulnar axial loading model was adopted, to which a 3-15 N compressive load was applied, resulting in approximately 440-3200 μɛ of compression in the bone surface. In the loaded ulnae, the degree of preferential apatite c-axis orientation along the ulnar long axis increased in a dose-dependent manner up to 13 N, whereas the increase in BMD was not dose-dependent. The Young's modulus along the same direction was enhanced as a function of the degree of apatite orientation. This finding indicates that bone has a mechanism that modifies the directionality (anisotropy) of its microstructure, strengthening itself specifically in the loaded direction. BMD, a scalar quantity, does not allow for load-direction-specific strengthening. Functional adaptation through changes in apatite orientation is an excellent strategy for bones to efficiently change their strength in response to external loading, which is mostly anisotropic.

The auditory brainstem response (ABR) is a scalp recording of potentials produced by sound stimulation, and is commonly used as an indicator of auditory function. However, the ABR threshold, which is the lowest audible sound pressure, cannot be objectively determined since it is determined visually using a measurer, and this has been a problem for several decades. Although various algorithms have been developed to objectively determine ABR thresholds, they remain lacking in terms of accuracy, efficiency, and convenience. Accordingly, we proposed an improved algorithm based on the mutual covariance at adjacent sound pressure levels. An ideal ABR waveform with clearly defined waves I–V was created; moreover, using this waveform as a standard template, the experimentally obtained ABR waveform was inspected for disturbances based on mutual covariance. The ABR testing was repeated if the value was below the established cross-covariance reference value. Our proposed method allowed more efficient objective determination of ABR thresholds and a smaller burden on experimental animals.