A bone substitute with high affinity for vitamin D-binding protein―relationship with niche of osteoclasts

The biological activity of osteoblasts and osteoclasts is regulated not only by hormones but also by local growth factors, which are expressed in neighbouring cells or included in bone matrix. Previously, we developed hydroxyapatite (HA) composed of rod-shaped particles using applied hydrothermal methods (HHA), and it revealed mild biodegradability and potent osteoclast homing activity. Here, we compared serum proteins adsorbed to HHA with those adsorbed to conventional HA composed of globular-shaped particles (CHA). The two ceramics adsorbed serum albumin and γ-globulin to similar extents, but affinity for γ-globulin was much greater than that to serum albumin. The chemotactic activity for macrophages of serum proteins adsorbed to HHA was significantly higher than that of serum proteins adsorbed to CHA. Quantitative proteomic analysis of adsorbed serum proteins revealed preferential binding of vitamin D-binding protein (DBP) and complements C3 and C4B with HHA. When implanted with the femur of 8-week-old rats, HHA contained significantly larger amount of DBP than CHA. The biological activity of DBP was analysed and it was found that the chemotactic activity for macrophages was weak. However, DBP-macrophage activating factor, which is generated by the digestion of sugar chains of DBP, stimulated osteoclastogenesis. These results confirm that the microstructure of hydroxyapatite largely affects the affinity for serum proteins, and suggest that DBP preferentially adsorbed to HA composed of rod-shaped particles influences its potent osteoclast homing activity and local bone metabolism

Osteoconductivity of hydrothermally synthesized beta-tricalcium phosphate composed of rod-shaped particles under mechanical unloading

Spherical beta-tricalcium phosphate (β-TCP) granules synthesized using a unique dropping slurry method expressed good osteoconductivity with prominent bone apposition and bioresorbability when implanted into the rat femur (Gonda et al, Key Eng. Mater. 361-363:1013-1016, 2008). The spherical β-TCP granules were implanted into the bone defect created in the distal end of the right femur of each 8-week-old female Wistar rat. To analyze performance of the spherical β-TCP granules as bone substitute in the bone with reduction in osteogenic potential, the right sciatic neurectomy was performed after implantation and the right hind limb was kept unloaded for 2 weeks before euthanization. Four weeks after implantation, some spherical β-TCP granules with resorption in part were surrounded by newly formed bone. Eight and 12 weeks after implantation, most of the residual β-TCP granules were embedded in newly formed bone, and total volume of the implant and newly formed bone was more than the other portions of the bone or the bone of control animals. Osteoclast activity in the implanted area was also higher than the other portions of the bone or the bone of control animals. Replacement of the intraosseous residual β-TCP granules for bone progressed at 12 weeks after implantation compared to those at 8 weeks after implantation. These data suggested that the spherical β-TCP granules stimulated osteogenesis and osteoclast activity of the unloaded bone

Stimulatory effect of hydrothermally synthesized biodegradable hydroxyapatite granules on osteogenesis and direct association with osteoclasts.

Calcium-deficient hydroxyapatite (HA) granules with a unique spherical shape were prepared using an applied hydrothermal method. Spherical stoichiometric HA granules were also prepared by normal sintering and both granules were used for implantation into rat tibiae to compare the biological responses to each implant. Twelve and 24 weeks after implantation, the volume of calcium-deficient HA granules was significantly less than that of stoichiometric HA granules, and the biodegradability of calcium-deficient HA granules was confirmed. The larger number of osteoclasts, larger osteoblast surface and larger bone volume in the implanted area of calcium-deficient HA than those of stoichiometric HA suggested that osteoclastic resorption of calcium-deficient HA affected osteogenesis in that area. To analyze the direct contribution of osteoclasts to osteogenesis, C2C12 multipotent myoblastic cells, which have the potential to differentiate into osteoblasts in the presence of bone morphogenetic protein 2, were cultured with supernatants of osteoclasts cultured on calcium-deficient HA, stoichiometric HA, beta-tricalcium phosphate disks or plastic dishes, or bone marrow macrophages cultured on plastic dishes. Supernatants of osteoclasts but not bone marrow macrophages stimulated the expression of Runx2 and osteocalcin in C2C12 cells in concert with bone morphogenetic protein 2. The expression of alkaline phosphatase was stimulated with supernatants of osteoclasts cultured on ceramic disks. These results suggested that osteoclasts produced certain soluble factors which stimulated osteoblastic differentiation and they were thought to be associated with the induction of a larger osteoblast surface and bone volume in the animals implanted with calcium-deficient HA granules

SALC ノ ガクセイ スタッフ ト ツクル ニホンゴ ガクシュウ シエン OU マルチリンガルプラザ ノ カツドウ カラ

実践報告Activity Reports本稿は、大阪大学国際教育交流センター日本語教育研究チームで取り組んできた日本語学習支援の実践報告である。2023年度の特筆すべき事項として、大阪大学のSALC（Self-Access Learning Center）であるOU マルチリンガルプラザのこれまでの活動に基づき、大学院生スタッフが従事しやすい体制を整備したことが挙げられる。SALC の運営は、それを運営している教育機関や学生の特色に合った体制を設計することにより、より機能的なものとなる。本稿では、大阪大学の日本語学習者とOU マルチリンガルプラザで従事する学生スタッフの概要を説明したのち、これまでの活動経緯とその成果を報告し、今後の課題を述べる