Review : Research and Development of Titanium-Containing Biomedical High Entropy Alloys (BioHEAs) Utilizing Rapid Solidification via Laser-Powder Bed Fusion

High entropy alloys (HEAs) have been developed as a new class of structural materials that consist of multicomponent elements with an approximately equiatomic ratio for increasing the mixing entropy to stabilize the solid solution phase. HEA for biomedical applications (BioHEA) was first developed in Japan; HEA comprising nonbiotoxic elements was specifically designed, demonstrating excellent mechanical properties and biocompatibility. However, elemental segregation, often observed in BioHEAs, hinders the inherent functions derived from high entropy effects and solid solution hardening. In this review article, elemental homogenization and functionalization of BioHEAs utilized by ultra-rapid cooling via laser-powder bed fusion and the characteristics of these BioHEAs, especially focusing on their excellent properties for biomedical applications, are introduced.Ozasa Ryosuke, Matsugaki Aira, Ishimoto Takuya, et al. Review : Research and Development of Titanium-Containing Biomedical High Entropy Alloys (BioHEAs) Utilizing Rapid Solidification via Laser-Powder Bed Fusion. MATERIALS TRANSACTIONS 64, 31 (2023); https://doi.org/10.2320/matertrans.MT-MLA2022011

Ibandronate Suppresses Changes in Apatite Orientation and Young's Modulus Caused by Estrogen Deficiency in Rat Vertebrae

Bone material quality is important for evaluating the mechanical integrity of diseased and/or medically treated bones. However, compared to the knowledge accumulated regarding changes in bone mass, our understanding of the quality of bone material is lacking. In this study, we clarified the changes in bone material quality mainly characterized by the preferential orientation of the apatite c-axis associated with estrogen deficiency-induced osteoporosis, and their prevention using ibandronate (IBN), a nitrogen-containing bisphosphonate. IBN effectively prevented bone loss and degradation of whole bone strength in a dose-dependent manner. The estrogen-deficient condition abnormally increased the degree of apatite orientation along the craniocaudal axis in which principal stress is applied; IBN at higher doses played a role in maintaining the normal orientation of apatite but not at lower doses. The bone size-independent Young's modulus along the craniocaudal axis of the anterior cortical shell of the vertebra showed a significant and positive correlation with apatite orientation; therefore, the craniocaudal Young’s modulus abnormally increased under estrogen-deficient conditions, despite a significant decrease in volumetric bone mineral density. However, the abnormal increase in craniocaudal Young's modulus did not compensate for the degradation of whole bone mechanical properties due to the bone loss. In conclusion, it was clarified that changes in the material quality, which are hidden in bone mass evaluation, occur with estrogen deficiency-induced osteoporosis and IBN treatment. Here, IBN was shown to be a beneficial drug that suppresses abnormal changes in bone mechanical integrity caused by estrogen deficiency at both the whole bone and material levels.Ishimoto T., Saito M., Ozasa R., et al. Ibandronate Suppresses Changes in Apatite Orientation and Young's Modulus Caused by Estrogen Deficiency in Rat Vertebrae. Calcified Tissue International, 110, 6, 736. https://doi.org/10.1007/s00223-021-00940-2

Combination treatment with ibandronate and eldecalcitol prevents osteoporotic bone loss and deterioration of bone quality characterized by nano-arrangement of the collagen/apatite in an ovariectomized aged rat model

Combination therapy with bisphosphonates and vitamin D3 analogs has been frequently used for the treatment of osteoporosis. However, its effects on bone anisotropies, such as orientations of collagen and apatite at the nanometer-scale, which is a promising bone quality index, and its trabecular architecture at the micrometer scale, are not well understood despite its important mechanical properties and its role in fracture risk. In the present study, we analyzed the effects of ibandronate (IBN), eldecalcitol (ELD), and their combination on the collagen/apatite orientation and trabecular architectural anisotropy using an estrogen-deficiency-induced osteoporotic rat model. Estrogen deficiency caused by ovariectomy (OVX) excessively increased the degree of collagen/apatite orientation or trabecular architectural anisotropy along the craniocaudal axis in the lumbar vertebra compared to that of the sham-operated group. The craniocaudal axis corresponds to the direction of principal stress in the spine. The excessive material anisotropy in the craniocaudal axis contributed to the enhanced Young's modulus, which may compensate for the reduced mechanical resistance by bone loss to some extent. The solo administration of IBN and ELD prevented the reduction of bone fraction (BV/TV) determined by μ-CT, and combination therapy showed the highest efficacy in BV/TV gain. Furthermore, the solo administration and combination treatment significantly decreased the degree of collagen/apatite orientation to the sham level. Based on the results of bone mass and collagen/apatite orientation, combination treatment is an effective strategy. This is the first report to demonstrate the efficacy of IBN, ELD, and combination treatment with IBN and ELD relative to the bone micro-architectural anisotropy characterized by collagen/apatite orientation.Ozasa R., Saito M., Ishimoto T., et al. Combination treatment with ibandronate and eldecalcitol prevents osteoporotic bone loss and deterioration of bone quality characterized by nano-arrangement of the collagen/apatite in an ovariectomized aged rat model. Bone, 157, 116309. https://doi.org/10.1016/j.bone.2021.116309

A Novel Ex Vivo Bone Culture Model for Regulation of Collagen/Apatite Preferential Orientation by Mechanical Loading

The anisotropic microstructure of bone, composed of collagen fibers and biological apatite crystallites, is an important determinant of its mechanical properties. Recent studies have revealed that the preferential orientation of collagen/apatite composites is closely related to the direction and magnitude of in vivo principal stress. However, the mechanism of alteration in the collagen/apatite microstructure to adapt to the mechanical environment remains unclear. In this study, we established a novel ex vivo bone culture system using embryonic mouse femurs, which enabled artificial control of the mechanical environment. The mineralized femur length significantly increased following cultivation; uniaxial mechanical loading promoted chondrocyte hypertrophy in the growth plates of embryonic mouse femurs. Compressive mechanical loading using the ex vivo bone culture system induced a higher anisotropic microstructure than that observed in the unloaded femur. Osteocytes in the anisotropic bone microstructure were elongated and aligned along the long axis of the femur, which corresponded to the principal loading direction. The ex vivo uniaxial mechanical loading successfully induced the formation of an oriented collagen/apatite microstructure via osteocyte mechano-sensation in a manner quite similar to the in vivo environment.Watanabe R., Matsugaki A., Ishimoto T., et al. A Novel Ex Vivo Bone Culture Model for Regulation of Collagen/Apatite Preferential Orientation by Mechanical Loading. International Journal of Molecular Sciences, 23, 13, 7423. https://doi.org/10.3390/ijms23137423

Construction of human induced pluripotent stem cell-derived oriented bone matrix microstructure by using in vitro engineered anisotropic culture model

Bone tissue has anisotropic microstructure based on collagen/biological apatite orientation, which plays essential roles in the mechanical and biological functions of bone. However, obtaining an appropriate anisotropic microstructure during the bone regeneration process remains a great challenging. A powerful strategy for the control of both differentiation and structural development of newly-formed bone is required in bone tissue engineering, in order to realize functional bone tissue regeneration. In this study, we developed a novel anisotropic culture model by combining human induced pluripotent stem cells (hiPSCs) and artificially-controlled oriented collagen scaffold. The oriented collagen scaffold allowed hiPSCs-derived osteoblast alignment and further construction of anisotropic bone matrix which mimics the bone tissue microstructure. To the best of our knowledge, this is the first report showing the construction of bone mimetic anisotropic bone matrix microstructure from hiPSCs. Moreover, we demonstrated for the first time that the hiPSCs-derived osteoblasts possess a high level of intact functionality to regulate cell alignment. © 2017 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 360–369, 2018.Ozasa R., Matsugaki A., Isobe Y., et al. Construction of human induced pluripotent stem cell-derived oriented bone matrix microstructure by using in vitro engineered anisotropic culture model. Journal of Biomedical Materials Research - Part A, 106, 2, 360. https://doi.org/10.1002/jbm.a.36238

Superior alignment of human ipsc-osteoblasts associated with focal adhesion formation stimulated by oriented collagen scaffold

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.Ozasa R., Matsugaki A., Matsuzaka T., et al. Superior alignment of human ipsc-osteoblasts associated with focal adhesion formation stimulated by oriented collagen scaffold. International Journal of Molecular Sciences, 22, 12, 6232. https://doi.org/10.3390/ijms22126232

Factor which governs the feature of texture developed during additive manufacturing; clarified from the study on hexagonal C40-NbSi2

C40-NbSi2 with a hexagonal unit cell is focused as a high-temperature structural material. We first attempted the fabrication of the bulk C40-NbSi2 products via selective laser melting (SLM) in additive manufacturing (AM) process. Strong crystallographic texture control wherein was parallel to the building direction, i.e. development of the so-called basal fiber texture, was achieved in this study. The texture developed in products does not largely vary by changing the scanning strategy, unlike the textures of C11b-MoSi2 with a tetragonal unit cell and a β-Ti alloy with a cubic unit cell. A comparison of these results led us to the conclusion that crystal symmetry, i.e., the multiplicity of the preferential crystal growth direction, is one of the primary factors that governs the features of the textures developed in AM-built materials.Hagihara K., Ishimoto T., Suzuki M., et al. Factor which governs the feature of texture developed during additive manufacturing; clarified from the study on hexagonal C40-NbSi2. Scripta Materialia, 203, 114111. https://doi.org/10.1016/j.scriptamat.2021.114111

Osteoporosis Changes Collagen/Apatite Orientation and Young’s Modulus in Vertebral Cortical Bone of Rat

This study revealed the distinguished changes of preferential orientation of collagen and apatite and Young’s modulus in two different types of osteoporotic bones compared with the normal bone. Little is known about the bone material properties of osteoporotic bones; therefore, we aimed to assess material properties in osteoporotic bones. 66 female Sprague–Dawley rats were used. We analyzed the volumetric bone mineral density, collagen/apatite orientation, and Young’s modulus of fifth lumbar vertebral cortex for osteoporotic rats caused by ovariectomy (OVX), administration of low calcium and phosphate content (LCaP) diet, and their combination (OVX + LCaP), as well as sham-operated control. Osteocyte conditions were assessed by hematoxylin and eosin and immunohistochemical (matrix extracellular phosphoglycoprotein (MEPE) and dentin matrix protein 1 (DMP1)) staining. All osteoporotic animals showed bone loss compared with the sham-operated control. OVX improved craniocaudal Young’s modulus by enhancing collagen/apatite orientation along the craniocaudal axis, likely in response to the elevated stress due to osteoporotic bone loss. Conversely, LCaP-fed animals showed either significant bone loss or degraded collagen/apatite orientation and Young’s modulus. Osteocytes in LCaP and OVX + LCaP groups showed atypical appearance and MEPE- and DMP1-negative phenotype, whereas those in the OVX group showed similarity with osteocytes in the control group. This suggests that osteocytes are possibly involved in the osteoporotic changes in collagen/apatite orientation and Young’s modulus. This study is the first to demonstrate that osteoporosis changes collagen/apatite orientation and Young’s modulus in an opposite manner depending on the cause of osteoporosis in spite of common bone loss.Ozasa R., Ishimoto T., Miyabe S., et al. Osteoporosis Changes Collagen/Apatite Orientation and Young’s Modulus in Vertebral Cortical Bone of Rat. Calcified Tissue International, 104, 4, 449. https://doi.org/10.1007/s00223-018-0508-z

Effect of a helium gas atmosphere on the mechanical properties of Ti-6Al-4V alloy built with laser powder bed fusion: A comparative study with argon gas

In metal additive manufacturing, the microstructures and associated mechanical properties of metal specimens can be controlled over a wide range. Although process parameters are considered important in the fabrication of functional parts, the effect of atmospheric gas has not been comprehensively documented. In laser powder bed fusion (LPBF), gas flow is used to eliminate fumes generated by laser irradiation. Simultaneously, the gas removes heat from the laser-irradiated part, which is exposed to high temperature. In this study, we investigated the capacity of helium as an alternative to argon, which is conventionally used as the LPBF atmosphere gas. He has a higher thermal conductivity and lower gas density than Ar, which may result in enhanced heat removal from the Ti-6Al-4V alloy during fabrication. Numerical simulations suggest a greater cooling rate under He flow. Further, the material built under He flow contained finer α' martensite grains and showed improved mechanical properties compared to those fabricated under Ar flow, despite the identical laser irradiation conditions. Thus, He gas is advantageous in LPBF for fabricating products with superior mechanical performance through microstructural refinement, and this is a result of its capacity for cooling and fume generation inhibition. Therefore, this study reveals the importance of the choice of atmospheric gas because of its effects on the characteristics of metallic specimens fabricated using LPBF.Amano H., Ishimoto T., Suganuma R., et al. Effect of a helium gas atmosphere on the mechanical properties of Ti-6Al-4V alloy built with laser powder bed fusion: A comparative study with argon gas. Additive Manufacturing, 48, 102444. https://doi.org/10.1016/j.addma.2021.102444

Host bone microstructure for enhanced resistance to bacterial infections

Postoperative bacterial infection is a serious complication of orthopedic surgery. Not only infections that develop in the first few weeks after surgery but also late infections that develop years after surgery are serious problems. However, the relationship between host bone and infection activation has not yet been explored. Here, we report a novel association between host bone collagen/apatite microstructure and bacterial infection. The bone-mimetic-oriented micro-organized matrix structure was obtained by prolonged controlled cell alignment using a grooved-structured biomedical titanium alloy. Surprisingly, we have discovered that highly aligned osteoblasts have a potent inhibitory effect on Escherichia coli adhesion. Additionally, the oriented collagen/apatite micro-organization of the bone matrix showed excellent antibacterial resistance against Escherichia coli. The proposed mechanism for realizing the antimicrobial activity of the micro-organized bone matrix is by the controlled secretion of the antimicrobial peptides, including β-defensin 2 and β-defensin 3, from the highly aligned osteoblasts. Our findings contribute to the development of anti-infective strategies for orthopedic surgeries. The recovery of the intrinsically ordered bone matrix organization provides superior antibacterial resistance after surgery.Watanabe R., Matsugaki A., Gokcekaya O., et al. Host bone microstructure for enhanced resistance to bacterial infections. Biomaterials Advances 154, 213633 (2023); https://doi.org/10.1016/j.bioadv.2023.213633