研究者詳細 - 松本　卓也

論文 - 松本　卓也

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Optimizing β-TCP with E-rhBMP-2-infused fibrin for vertical bone regeneration in a mouse calvarium model

Kun Zhao, Mitsuaki Ono, Xindi Mu, Ziyi Wang, Shichao Xie, Tomoko Yonezawa, Masahiro Okada, Takuya Matsumoto, Takuo Kuboki, Toshitaka Oohashi

Regenerative Biomaterials 12 2025年1月

掲載種別：研究論文（学術雑誌）出版者・発行元：Oxford University Press (OUP)

Abstract

Effective reconstruction of large bone defects, particularly in thickness, remains one of the major challenges in orthopedic and dental fields. We previously produced an Escherichia coli-based industrial-scale GMP-grade recombinant human bone morphogenetic protein-2 (E-rhBMP-2) and showed that the combination of E-rhBMP-2 with beta-tricalcium phosphate (β-TCP/E-rhBMP-2) can effectively promote bone reconstruction. However, the limited mechanical strength and poor morphology retention of β-TCP granules are key points that need optimization to obtain more effective grafts and further expand its clinical applications. Therefore, we combined β-TCP/E-rhBMP-2 with fibrin gel to enhance its mechanical properties and usability for vertical bone regeneration. We investigated the mechanical properties and vertical bone regeneration effects of the materials applied, with or without fibrin containing E-rhBMP-2, in a calvarial defect model in mice. Compression tests were conducted to assess the initial stability of the materials. Scanning electron microscopy and Fourier transform infrared spectroscopy were conducted to characterize the presence of fibrin on the scaffold. After 4 and 12 weeks of implantation, micro-computed tomography and histological and immunofluorescent analyses were performed to assess the morphology and volume of the newly formed bone. The fibrin-containing groups had significantly higher initial mechanical strength and higher ability to maintain their morphology in vivo compared to the counterparts without fibrin. However, fibrin gel alone suppressed the bone formation ability of β-TCP/E-rhBMP-2 whereas the presence of high doses of E-rhBMP-2 in fibrin gel resulted in material resorption and enhanced new bone formation. In conclusion, fibrin gel significantly improved the mechanical strength and surgical manageability of the β-TCP/E-rhBMP-2 scaffold, and the addition of E-rhBMP-2 to the fibrin gel further enhanced the vertical bone regeneration and initial structural integrity of the scaffold.

DOI： 10.1093/rb/rbae144

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その他リンク： https://academic.oup.com/rb/article-pdf/doi/10.1093/rb/rbae144/61841803/rbae144.pdf

Long-Term Bonding Performance of One-Bottle vs. Two-Bottle Bonding Agents to Lithium Disilicate Ceramics 査読

Masao Irie, Masahiro Okada, Yukinori Maruo, Goro Nishigawa, Takuya Matsumoto

Polymers 16 ( 16 ) 2266 - 2266 2024年8月

掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

The aim of this study was to compare the long-term bonding performance to lithium disilicate (LDS) ceramic between one-bottle and two-bottle bonding agents. Bonding performance was investigated under these LDS pretreatment conditions: with hydrofluoric acid (HF) only, without HF, with a two-bottle bonding agent (Tokuyama Universal Bond II) only. Shear bond strengths between LDS and nine resin cements (both self-adhesive and conventional adhesive types) were measured at three time periods: after one-day water storage (Base), and after 5000 and 20,000 thermocycles (TC 5k and TC 20k respectively). Difference in degradation between one- and two-bottle bonding agents containing the silane coupling agent was compared by high-performance liquid chromatography. With HF pretreatment, bond strengths were not significantly different among the three time periods for each resin cement. Without HF, ESTECEM II and Super-Bond Universal showed significantly higher values than others at TC 5k and TC 20k when treated with the recommended bonding agents, especially at TC 20k. Difference in degradation between one- and two-bottle bonding agents containing the silane coupling agent was compared by high-performance liquid chromatography (HPLC). For both cements, these values at TC 20k were also not significantly different from pretreatment with only Tokuyama Universal Bond II. For LDS, long-term bond durability could be maintained by pretreatment with Tokuyama Universal Bond II instead of the hazardous HF.

DOI： 10.3390/polym16162266

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The cell membrane as biofunctional material for accelerated bone repair. 査読国際誌

Emi Hatano, Nahid Akhter, Risa Anada, Mitsuaki Ono, Toshitaka Oohashi, Takuo Kuboki, Hiroshi Kamioka, Masahiro Okada, Takuya Matsumoto, Emilio Satoshi Hara

Acta biomaterialia 2024年7月

記述言語：英語掲載種別：研究論文（学術雑誌）

The cell (plasma) membrane is enriched with numerous receptors, ligands, enzymes, and phospholipids that play important roles in cell-cell and cell-extracellular matrix interactions governing, for instance, tissue development and repair. We previously showed that plasma membrane nanofragments (PMNFs) act as nucleation sites for bone formation in vivo, and induce in vitro mineralization within 1 day. In this study, we optimized the methods for generating, isolating, and applying PMNFs as a cell-free therapeutic to expedite bone defect repair. The PMNFs were isolated from different mouse cell lines (chondrocytes, osteoblasts, and fibroblasts), pre-conditioned, lyophilized, and subsequently transplanted into 2 mm critical-sized calvarial defects in mice (n = 75). The PMNFs from chondrocytes, following a 3-day pre-incubation, significantly accelerated bone repair within 2 weeks, through a coordinated attraction of macrophages, endothelial cells, and osteoblasts to the healing site. In vitro experiments confirmed that PMNFs enhanced cell adhesion. Comparison of the PMNF efficacy with phosphatidylserine, amorphous calcium phosphate (ACP), and living cells confirmed the unique ability of PMNFs to promote accelerated bone repair. Importantly, PMNFs promoted nearly complete integration of the regenerated bone with native tissue after 6 weeks (% non-integrated bone area = 15.02), contrasting with the partial integration (% non-integrated bone area = 56.10; p < 0.01, Student's test) with transplantation of ACP. Vickers microhardness tests demonstrated that the regenerated bone after 6 weeks (30.10 ± 1.75) exhibited hardness similar to native bone (31.07 ± 2.46). In conclusion, this is the first study to demonstrate that cell membrane can be a promising cell-free material with multifaceted biofunctional properties that promote accelerated bone repair. STATEMENT OF SIGNIFICANCE.

DOI： 10.1016/j.actbio.2024.07.037

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Water-Mediated On-Demand Detachable Solid-State Adhesive of Porous Hydroxyapatite for Internal Organ Retractions. 査読国際誌

Masahiro Okada, Shi Chao Xie, Yusuke Kobayashi, Hiroaki Yanagimoto, Daisuke Tsugawa, Masaru Tanaka, Takayoshi Nakano, Takumi Fukumoto, Takuya Matsumoto

Advanced healthcare materials e2304616 2024年5月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Novel adhesives for biological tissues offer an advanced surgical approach. Here, the authors report the development and application of solid-state adhesives consisting of porous hydroxyapatite (HAp) biocompatible ceramics as novel internal organ retractors. The operational principles of the porous solid-state adhesives are experimentally established in terms of water migration from biological soft tissues into the pores of the adhesives, and their performance is evaluated on several soft tissues with different hydration states. As an example of practical medical utility, HAp adhesive devices demonstrate the holding ability of porcine livers and on-demand detachability in vivo, showing great potential as internal organ retractors in laparoscopic surgery.

DOI： 10.1002/adhm.202304616

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Lipids and Minerals, Interplay in Biomineralization: Nature's Alchemy 査読

Bhingaradiya Nutan, Masahiro Okada, Takuya Matsumoto

Tissue Engineering Part B: Reviews 2024年4月

Adaptive enhancement of apatite crystal orientation and Young's modulus under elevated load in rat ulnar cortical bone 査読

Jun Wang, Takuya Ishimoto, Tadaaki Matsuzaka, Aira Matsugaki, Ryosuke Ozasa, Takuya Matsumoto, Mikako Hayashi, Hyoung Seop Kim, Takayoshi Nakano

Bone 181 2024年4月

掲載種別：研究論文（学術雑誌）

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.

DOI： 10.1016/j.bone.2024.117024

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Mid-infrared passive spectroscopic imaging for visualizing tooth quality 査読

So Yamashita, Masahiro Okada, Takuya Matsumoto, Ichiro Ishimaru

Journal of Materials Chemistry B 2024年

Solid-state inorganic and metallic adhesives for soft biological tissues 査読

Masahiro Okada, Takuya Matsumoto

Japanese Dental Science Review 59 439 - 445 2023年12月

Fabrication of initial trabecular bone inspired three-dimensional structure with cell membrane nanofragments 査読

Koichi Kadoya, Emilio Satoshi Hara, Masahiro Okada, Yu Yang Jiao, Takayoshi Nakano, Akira Sasaki, Takuya Matsumoto

Regenerative Biomaterials 10 rbac088 2023年12月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Oxford University Press (OUP)

Abstract

The extracellular matrix of trabecular bone has a large surface exposed to the bone marrow and plays important roles such as hematopoietic stem cell niche formation and maintenance. In vitro reproduction of trabecular bone microenvironment would be valuable not only for developing a functional scaffold for bone marrow tissue engineering but also for understanding its biological functions. Herein, we analyzed and reproduced the initial stages of trabecular bone formation in mouse femur epiphysis. We identified that the trabecular bone formation progressed through the following steps: (1) partial rupture of hypertrophic chondrocytes; (2) calcospherite formation on cell nanofragments (CNFs) derived from the ruptured cell membranes; and (3) calcospherite growth and fusion to form the initial three-dimensional (3D) structure of trabecular bones. For reproducing the initial trabecular bone formation in vitro, we collected CNFs from cultured cells and used as nucleation sites for biomimetic calcospherite formation. Strikingly, almost the same 3D structure of the initial trabecular bone could be obtained in vitro by using additional CNFs as a binder to fuse biomimetic calcospherites.

DOI： 10.1093/rb/rbac088

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Investigation on bacterial capture and antibacterial properties of acid-treated Ti surface 査読

Haruyuki Aoyagi, Masahiro Okada, Hiroaki Yanagimoto, Takuya Matsumoto

Dental Materials 2023年12月

Fabrication of a Fish-Bone-Inspired Inorganic–Organic Composite Membrane 査読

YuYang Jiao, Masahiro Okada, Bhingaradiya Nutan, Noriyuki Nagaoka, Ahmad Bikharudin, Randa Musa, Takuya Matsumoto

Polymers 15 ( 20 ) 4190 - 4190 2023年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

Biological materials have properties like great strength and flexibility that are not present in synthetic materials. Using the ribs of crucian carp as a reference, we investigated the mechanisms behind the high mechanical properties of this rib bone, and found highly oriented layers of calcium phosphate (CaP) and collagen fibers. To fabricate a fish-rib-bone-mimicking membrane with similar structure and mechanical properties, this study involves (1) the rapid synthesis of plate-like CaP crystals, (2) the layering of CaP–gelatin hydrogels by gradual drying, and (3) controlling the shape of composite membranes using porous gypsum molds. Finally, as a result of optimizing the compositional ratio of CaP filler and gelatin hydrogel, a CaP filler content of 40% provided the optimal mechanical properties of toughness and stiffness similar to fish bone. Due to the rigidity, flexibility, and ease of shape control of the composite membrane materials, this membrane could be applied as a guided bone regeneration (GBR) membrane.

DOI： 10.3390/polym15204190

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歯根膜応用を目指した石灰化dECM膜のin vivo骨結合性評価

鈴木美加, 木村剛, 岡田正弘, 松本卓也, 山田将博, 江草宏, 中村奈緒子, 岸田晶夫

日本歯科理工学会誌 42 ( Special Issue81 ) 18 - 18 2023年3月

Preparation of mineralized pericardium by alternative soaking for soft–hard interregional tissue application 査読

Mika Suzuki, Tsuyoshi Kimura, Yuta Nakano, Mako Kobayashi, Masahiro Okada, Takuya Matsumoto, Naoko Nakamura, Yoshihide Hashimoto, Akio Kishida

Journal of Biomedical Materials Research Part A 111 ( 2 ) 198 - 208 2023年2月

Shear Bond Strength of Resin Luting Materials to Lithium Disilicate Ceramic: Correlation between Flexural Strength and Modulus of Elasticity 査読

Masao Irie, Masahiro Okada, Yukinori Maruo, Goro Nishigawa, Takuya Matsumoto

Polymers 15 ( 5 ) 1128 - 1128 2023年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

This study investigates the effect of the curing mode (dual-cure vs. self-cure) of resin cements (four self-adhesive and seven conventional cements) on their flexural strength and flexural modulus of elasticity, alongside their shear bond strength to lithium disilicate ceramics (LDS). The study aims to determine the relationship between the bond strength and LDS, and the flexural strength and flexural modulus of elasticity of resin cements. Twelve conventional or adhesive and self-adhesive resin cements were tested. The manufacturer’s recommended pretreating agents were used where indicated. The shear bond strengths to LDS and the flexural strength and flexural modulus of elasticity of the cement were measured immediately after setting, after one day of storage in distilled water at 37 °C, and after 20,000 thermocycles (TC 20k). The relationship between the bond strength to LDS, flexural strength, and flexural modulus of elasticity of resin cements was investigated using a multiple linear regression analysis. For all resin cements, the shear bond strength, flexural strength, and flexural modulus of elasticity were lowest immediately after setting. A clear and significant difference between dual-curing and self-curing modes was observed in all resin cements immediately after setting, except for ResiCem EX. Regardless of the difference of the core-mode condition of all resin cements, flexural strengths were correlated with the LDS surface upon shear bond strengths (R2 = 0.24, n = 69, p < 0.001) and the flexural modulus of elasticity was correlated with them (R2 = 0.14, n = 69, p < 0.001). Multiple linear regression analyses revealed that the shear bond strength was 17.877 + 0.166, the flexural strength was 0.643, and the flexural modulus was (R2 = 0.51, n = 69, p < 0.001). The flexural strength or flexural modulus of elasticity may be used to predict the bond strength of resin cements to LDS.

DOI： 10.3390/polym15051128

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Important roles of odontoblast membrane phospholipids in early dentin mineralization 査読国際誌

Risa Anada, Emilio Satoshi Hara, Noriyuki Nagaoka, Masahiro Okada, Hiroshi Kamioka, Takuya Matsumoto

Journal of Materials Chemistry B 11 ( 3 ) 657 - 666 2023年1月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Royal Society of Chemistry (RSC)

The objective of this study was to first identify the timing and location of early mineralization of mouse first molar, and subsequently, to characterize the nucleation site for mineral formation...

DOI： 10.1039/d2tb02351b

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Host bone microstructure for enhanced resistance to bacterial infections 査読

Ryota Watanabe, Aira Matsugaki, Ozkan Gokcekaya, Ryosuke Ozasa, Takuya Matsumoto, Hiroyuki Takahashi, Hidekazu Yasui, Takayoshi Nakano

Biomaterials Advances 2023年

掲載種別：研究論文（学術雑誌）

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.

DOI： 10.1016/j.bioadv.2023.213633

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Self-adhesive Devices Made of Titanium for Biological Soft Tissue 査読

Masahiro Okada, Takuya Matsumoto

Materia Japan 61 ( 11 ) 760 - 764 2022年11月

Does Multifunctional Acrylate’s Addition to Methacrylate Improve Its Flexural Properties and Bond Ability to CAD/CAM PMMA Block? 査読国際誌

Yukinori Maruo, Kumiko Yoshihara, Masao Irie, Noriyuki Nagaoka, Takuya Matsumoto, Shogo Minagi

Materials 15 ( 21 ) 7564 - 7564 2022年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

This study investigated the effects of a multifunctional acrylate copolymer—Trimethylolpropane Triacrylate (TMPTA) and Di-pentaerythritol Polyacrylate (A-DPH)—on the mechanical properties of chemically polymerized acrylic resin and its bond strength to a CAD/CAM polymethyl methacrylate (PMMA) disk. The methyl methacrylate (MMA) samples were doped with one of the following comonomers: TMPTA, A-DPH, or Trimethylolpropane Trimethacrylate (TMPTMA). The doping ratio ranged from 10 wt% to 50 wt% in 10 wt% increments. The flexural strength (FS) and modulus (FM) of PMMA with and without comonomer doping, as well as the shear bond strength (SBS) between the comonomer-doped PMMA and CAD/CAM PMMA disk, were evaluated. The highest FS (93.2 ± 4.2 MPa) was obtained when doped with 20 wt% of TMPTA. For TMPTMA, the FS decreased with the increase in the doping ratio. For SBS, TMPTA showed almost constant values (ranging from 7.0 to 8.2 MPa) regardless of the doping amount, and A-DPH peaked at 10 wt% doping (8.7 ± 2.2 MPa). TMPTMA showed two peaks at 10 wt% (7.2 ± 2.6 MPa) and 40 wt% (6.5 ± 2.3 MPa). Regarding the failure mode, TMPTMA showed mostly adhesive failure between the CAD/CAM PMMA disk and acrylic resin while TMPTA and A-DPH showed an increased rate of cohesive or mixed failures. Acrylate’s addition as a comonomer to PMMA provided improved mechanical properties and bond strength to the CAD/CAM PMMA disk.

DOI： 10.3390/ma15217564

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Distinct Morphologies of Bone Apatite Clusters in Endochondral and Intramembranous Ossification. 査読国際誌

Emilio Satoshi Hara, Noriyuki Nagaoka, Masahiro Okada, Takayoshi Nakano, Takuya Matsumoto

Advanced biology 6 ( 11 ) e2200076 2022年7月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Bone apatite crystals grow in clusters, but the microstructure of these clusters is unknown. This study compares the structural and compositional differences between bone apatite clusters formed in intramembranous (IO) and endochondral ossification (EO). Calvaria (IO) and femurs (EO) are isolated from mice at embryonic days (E) 14.5 to 15.5 and post-natal days (P) 6 to 7, respectively. Results show that the initially formed bone apatite clusters in EO (≅1.2 µm2 ) are >10 times larger than those in IO (≅0.1 µm2 ), without significant changes in ion composition. In IO (E14.5 calvarium), early minerals are formed inside matrix vesicles (MVs). In contrast, in EO (P6 femur epiphysis), no MVs are observed, and chondrocyte-derived plasma membrane nanofragments (PMNFs) are the nucleation site for mineralization. Apatite cluster size difference is linked with the different nucleation sites. Moreover, an alkaline pH and slow P supply into a Ca-rich microenvironment are suggested to facilitate apatite cluster growth, as demonstrated in a biomimetic mineralization system. Together, the results reveal for the first time the distinct and exquisite microstructures of bone apatite clusters in IO and EO, and provide insightful inspirations for the design of more efficient materials for bone tissue engineering and repair.

DOI： 10.1002/adbi.202200076

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

Ryota Watanabe, Aira Matsugaki, Takuya Ishimoto, Ryosuke Ozasa, Takuya Matsumoto, Takayoshi Nakano

International Journal of Molecular Sciences 23 ( 13 ) 7423 - 7423 2022年7月

掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

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.

DOI： 10.3390/ijms23137423

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