Homan Kentaro

Faculty of MedicineSpecially Appointed Assistant Professor
Last Updated :2025/06/07

■Researcher basic information

Degree

  • PhD, Hokkaido University

Researchmap personal page

Home Page URL

Researcher number

  • 40823331

ORCID ID

J-Global ID

Research Keyword

  • Regenerative medicine
  • osteoarthritis
  • orthopedics
  • biomechanics
  • gait
  • cartilage degradation
  • Rehabilitation medicine

Research Field

  • Life sciences, Molecular biology
  • Life sciences, Anatomy
  • Life sciences, Nutrition and health science
  • Life sciences, Physical and health education
  • Life sciences, Laboratory animal science
  • Life sciences, Biomaterials
  • Life sciences, Biomedical engineering
  • Life sciences, Medical equipment testing
  • Life sciences, Rehabilitation science
  • Life sciences, Orthopedics

■Research activity information

Awards

  • Oct. 2023, 第38回日本整形外科学会基礎学術集会優秀ポスター賞　　　　　　　　　　　　　　　
    山口純、小野寺智洋、宝満健太郎、松岡正剛、長濱宏治、上田菜摘美、澤田志穂、斎藤充、岩崎倫政
  • Jul. 2022, 第141回北海道整形災害外科学会学術集会最優秀発表賞　　　　　　　　　　　　　　　
    細川吉暁、小野寺智洋、宝満健太郎、工藤與亮、亀田浩之、杉森博行、岩崎倫政
  • Jul. 2017, 第133回北海道整形災害外科学会学術集会最優秀発表賞　　　　　　　　　　　　　　　
    松原新史、小野寺智洋、前田英次郎、門間太輔、松岡正剛、馬場力哉、本谷和俊、上徳善太、宝満健太郎、大橋俊朗、岩崎倫政
  • Jun. 2016, 第131回北海道整形災害外科学会学術集会最優秀発表賞　　　　　　　　　　　　　　　
    上徳善太、小野寺智洋、門間太輔、松岡正剛、馬場力哉、本谷和俊、松原新史、宝満健太郎、岩崎倫政

Papers

  • Tendon Tissue Regeneration With Cell Orientation Using an Injectable Alginate-Cell Cross-linked Gel.
    Jun Yamaguchi, Kentaro Homan, Tomohiro Onodera, Masatake Matsuoka, Shoutaro Arakawa, Natsumi Ueda, Shiho Sawada, Nana Kawate, Takayuki Nonoyama, Yoshinori Katsuyama, Koji Nagahama, Mitsuru Saito, Norimasa Iwasaki
    The American journal of sports medicine, 3635465251325498, 3635465251325498, 23 Mar. 2025, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, BACKGROUND: Tendons have a limited blood supply and form inferior scar tissue during repair, which increases the risk of reruptures, causes complications, and limits regenerative capacity. Current methods to repair injured tendon tissue use solid scaffolds, which carry the risk of contamination (infections) and require open surgery for transplantation. HYPOTHESIS: Alginate-cell cross-linked gels, which can be applied by a percutaneous injection and transmit mechanical stress to cells via direct cell interaction, could induce tendon tissue regeneration. STUDY DESIGN: Controlled laboratory study. METHODS: A cross-linked gel was prepared to suspend azide-modified mesenchymal stromal cells (MSCs) in a dibenzocyclooctyne-modified branched alginic acid solution. The cross-linked gel was cultured in a bioreactor. In vivo, the Achilles tendon defects of 104 Lewis rats were injected with saline (control group), alginate gel alone (alginate group), alginate gel with MSCs (MSC group), and cross-linked gel (cross-link group). At 2 and 4 weeks postoperatively, histological and biochemical evaluations were performed. The biomechanical properties of repaired tissue were assessed at 4 weeks. RESULTS: In the bioreactor culture, the cell orientation in the cross-linked gel was parallel to the direction of tension. Histological analysis of the cross-link group showed significantly more repaired tendon tissue and improved collagen fiber orientation compared with the alginate group or MSC group. The biomechanical properties of the cross-link group included higher stiffness. CONCLUSION: The cross-linked gel was injectable at the injury site and was able to induce tissue regeneration with cell-oriented adaptability to the mechanical environment of tissue defects. CLINICAL RELEVANCE: Intercellular cross-linking technology holds the potential for clinical application as a minimally invasive therapeutic approach that can contribute to the qualitative improvement of tendon tissue regeneration.
  • Intra-Articular Administration of Ganglioside Sugars Protects Cartilage from Progressive Degeneration in an Instability OA Rabbit Model.
    Masanari Hamasaki, Tomohiro Onodera, Junichi Furukawa, Masahiro Todoh, Yuma Sakai, Taku Ebata, Mohamad Alaa Terkawi, Kentaro Homan, Norimasa Iwasaki
    Cartilage, 19476035241311542, 19476035241311542, 14 Jan. 2025, [Peer-reviewed], [International Magazine]
    English, Scientific journal, OBJECTIVE: Osteoarthritis (OA) is a degenerative joint disease that has no cure, and current therapies are intended to minimize pain. There is, therefore, a need for effective pharmacologic agents that reverse or slow the progression of joint damage. We report herein on an investigation of the effects of intra-articular injections of ganglioside sugars on the progression of OA in an experimental rabbit model. DESIGN: Knee OA was induced Japanese in White rabbits by anterior cruciate ligament transection (ACLT). Ganglioside sugars at concentrations of 0.1, 0.3, and 0.9 mg/ml were then intra-articularly injected as a possible treatment for OA. Controls received intra-articular injections of saline. Knees were assessed macroscopically, histologically, and mechanically at 13 weeks after ACLT induction. RESULTS: Macroscopically, knees of the groups that received ganglioside sugars at concentrations of 0.3 and 0.9 mg/ml exhibited milder cartilage degradation compared to the controls. Consistent with these results, histological scores for these knees were significantly higher than the corresponding values for the control knees. Lectin histochemistry staining revealed that the treatment with ganglioside sugars at concentrations of 0.3 and 0.9 mg/ml was associated with a remarkable increase in the levels of GalNAc-positive chondrocytes in cartilage. Coefficient of friction testing also demonstrated that cartilages treated with ganglioside sugars had a lower coefficient of frictions than the values for the control group. CONCLUSIONS: Intra-articular injections of ganglioside sugars prevented cartilage degeneration in an OA-instability model. These results highlight the promising therapeutic potential for using ganglioside sugars in the treatment of progressive OA.
  • Chondroprotective functions of neutrophil-derived extracellular vesicles by promoting the production of secreted frizzled-related protein 5 in cartilage.
    Keita Kitahara, Taku Ebata, Chen Liyile, Yoshio Nishida, Yuki Ogawa, Taiki Tokuhiro, Junki Shiota, Tatsuya Nagano, Taichi E Takasuka, Tsutomu Endo, Tomohiro Shimizu, Hend Alhasan, Tsuyoshi Asano, Daisuke Takahashi, Kentaro Homan, Tomohiro Onodera, Ken Kadoya, M Alaa Terkawi, Norimasa Iwasaki
    Cell communication and signaling : CCS, 22, 1, 569, 569, 27 Nov. 2024, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: Osteoarthritis (OA) is the most common degenerative joint disease characterized by cartilage degradation and various degrees of inflammation in the synovium. Growing evidence highlights that neutrophil extracellular vesicles (EVs) play a protective role in arthritic joints by promoting the resolution of inflammation and the synthesis of proteoglycans in cartilage. However, this homeostatic function is dependent on the activation state of neutrophils and the surrounding environment/tissues. Hence, we explored the chondroprotective functions of neutrophil-derived EVs under different stimulation conditions and the underlying molecular mechanism. METHODS: Human blood-derived neutrophils, murine bone marrow-derived neutrophils, C-28I2 cells and primary chondrocytes were used. Neutrophils were stimulated with different cytokines, and their EVs were isolated for chondrocyte stimulation and further subjected to RNA-sequencing analysis. Two experimental murine OA models were used, and the treatment was performed by intraarticular injections. RESULTS: Conditioned medium from neutrophils stimulated with TGF-β (N-β) had the greatest inhibitory effect on the expression of catabolic factors in stimulated chondrocytes. These protective effects were not impaired when conditioned medium of N-β from AnxA1-deficient mice was used. Consistent with these results, EVs isolated from N-β significantly reduced the expression of catabolic factors in stimulated chondrocytes. Bulk RNA-seq analysis revealed that secreted frizzled-related protein 5 (SFRP5) is upregulated in N-β-EV-stimulated chondrocytes. Furthermore, recombinant SFRP5 treatment significantly reduced the expression of catabolic factors in vitro and catabolic process in experimental murine OA models. CONCLUSIONS: The current study emphasizes the potential therapeutic application of neutrophils in OA and provides new knowledge on the molecular mechanisms underlying their function.
  • Associations between glycan signature alterations and the cellular antigenic properties of passaged chondrocytes.
    Kentaro Homan, Taiki Tokuhiro, Tomohiro Onodera, Hisatoshi Hanamatsu, Jun-Ichi Furukawa, Taku Ebata, Masatake Matsuoka, Ken Kadoya, M Alaa Terkawi, Norimasa Iwasaki
    Frontiers in immunology, 15, 1475473, 1475473, 25 Nov. 2024, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, BACKGROUND: Cartilage repair is a significant clinical challenge because of the limited intrinsic healing capacity. Current therapeutic strategies, such as cell transplantation therapy, aim to overcome this challenge by replacing damaged tissue with healthy cells. However, the long-term survival and functionality of transplanted cells remain major hurdles. OBJECTIVE: This study investigated the impact of chondrocyte passaging on glycan profiles and their antigenic properties. We hypothesized that alterations in glycan composition due to passaging may contribute to the enhanced ability to activate macrophages, thereby affecting the outcome of cell transplantation therapy. METHODS: Peritoneal macrophages and primary articular chondrocytes were isolated from C57BL/6 mice to establish direct and indirect coculture models. Macrophage activation was assessed by measuring the concentrations of IL-6 and nitric oxide in the culture supernatants or their gene expression. Glycome analysis of various glycoconjugates was performed by glycoblotting methods combined with the SALSA procedure for N-glycans and GSLs and the BEP method for O-glycans. RESULTS: Our results revealed that direct coculture of macrophages with passaged chondrocytes increased the production of proinflammatory cytokines, including IL-6 and NO, as the number of passages increased. With increasing passage number, the expression of GD3 substantially decreased, and the expression of GM3, especially GD1a, significantly increased. Coculturing passaged GM3S knockout chondrocytes with macrophages significantly suppressed IL-6 expression, implying reduced macrophage activation. CONCLUSION: The observed activation of macrophages due to alterations in the glycan profile of chondrocytes provides a possible explanation for the antigenicity and immune rejection of transplanted cells.
  • CCR7 depletion alleviates bony growth imbalance following physeal injury in mice.
    WooYoung Kim, Yuko Sakai, Masatake Matsuoka, Yoshiaki Hosokawa, Ryuichi Fukuda, Kentaro Homan, Tomohiro Onodera, Norimasa Iwasaki
    Scientific reports, 14, 1, 24891, 24891, 22 Oct. 2024, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Growth plates are the frequent sites of skeletal injury in children, leading to skeletal growth imbalances. Chemokines, including the receptor CCR7, play a crucial role in stem cell recruitment and cartilage homeostasis, with previous studies linking CCR7 to osteoarthritis progression. However, its role in growth plate cartilage remains unclear. We analyzed the role of CCR7 in the physeal cartilage repair process in mice model. Physeal injury was created in the proximal tibia in 3-week-old C57BL/6 mice (WT) and CCR7-knockout mice (CCR7-/-). Tibial length was measured macroscopically and sections of the physeal injury were analyzed histologically and immunohistochemically. Height and bone volume of the tibial growth plate and bone mineral density (BMD) of the subchondral area were measured by micro-CT. Mesenchymal stem cells (MSCs) were harvested and gene expression after osteogenic differentiation was analyzed using qRT-PCR. At 1, 3 and 5 weeks postoperatively, injured tibiae of CCR7-/- mice were less shortened than those of WT mice. Bone volume of the physeal bridge was significantly lower in CCR7-/- mice than in WT mice. In contrast, BMD of the subchondral area was comparable between CCR-/- and WT mice, and between sham and operated tibiae. In osteogenic differentiation, CCR7-/- mice showed significantly lowered expression of osteogenic markers such as Osterix, Runx2 and Type X collagen. We demonstrated CCR7 depletion in mice inhibited physeal bridge formation and ameliorated growth imbalances after physeal injury.
  • Depletion of b-series ganglioside prevents limb length discrepancy after growth plate injury.
    Yoshiaki Hosokawa, Masatake Matsuoka, Yuko Sakai, Ryuichi Fukuda, Keizumi Matsugasaki, Kentaro Homan, Jun-Ichi Furukawa, Tomohiro Onodera, Norimasa Iwasaki
    BMC musculoskeletal disorders, 25, 1, 565, 565, 20 Jul. 2024, [Peer-reviewed], [International Magazine]
    English, Scientific journal, INTRODUCTION: Growth plate damage in long bones often results in progressive skeletal growth imbalance and deformity, leading to significant physical problems. Gangliosides, key glycosphingolipids in cartilage, are notably abundant in articular cartilage and regulate chondrocyte homeostasis. This suggests their significant roles in regulating growth plate cartilage repair. METHODS: Chondrocytes from 3 to 5 day-old C57BL/6 mice underwent glycoblotting and mass spectrometry. Based on the results of the glycoblotting analysis, we employed GD3 synthase knockout mice (GD3-/-), which lack b-series gangliosides. In 3-week-old mice, physeal injuries were induced in the left tibiae, with right tibiae sham operated. Tibiae were analyzed at 5 weeks postoperatively for length and micro-CT for growth plate height and bone volume at injury sites. Tibial shortening ratio and bone mineral density were measured by micro-CT. RESULTS: Glycoblotting analysis indicated that b-series gangliosides were the most prevalent in physeal chondrocytes among ganglioside series. At 3 weeks, GD3-/- exhibited reduced tibial shortening (14.7 ± 0.2 mm) compared to WT (15.0 ± 0.1 mm, P = 0.03). By 5 weeks, the tibial lengths in GD3-/- (16.0 ± 0.4 mm) closely aligned with sham-operated lengths (P = 0.70). Micro-CT showed delayed physeal bridge formation in GD3-/-, with bone volume measuring 168.9 ± 5.8 HU at 3 weeks (WT: 180.2 ± 3.2 HU, P = 0.09), but normalizing by 5 weeks. CONCLUSION: This study highlights that GD3 synthase knockout mice inhibit physeal bridge formation after growth plate injury, proposing a new non-invasive approach for treating skeletal growth disorders.
  • Glycosphingolipids in Osteoarthritis and Cartilage-Regeneration Therapy: Mechanisms and Therapeutic Prospects Based on a Narrative Review of the Literature
    Kentaro Homan, Tomohiro Onodera, Masatake Matsuoka, Norimasa Iwasaki
    International Journal of Molecular Sciences, 25, 9, 30 Apr. 2024, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, Glycosphingolipids (GSLs), a subtype of glycolipids containing sphingosine, are critical components of vertebrate plasma membranes, playing a pivotal role in cellular signaling and interactions. In human articular cartilage in osteoarthritis (OA), GSL expression is known notably to decrease. This review focuses on the roles of gangliosides, a specific type of GSL, in cartilage degeneration and regeneration, emphasizing their regulatory function in signal transduction. The expression of gangliosides, whether endogenous or augmented exogenously, is regulated at the enzymatic level, targeting specific glycosyltransferases. This regulation has significant implications for the composition of cell-surface gangliosides and their impact on signal transduction in chondrocytes and progenitor cells. Different levels of ganglioside expression can influence signaling pathways in various ways, potentially affecting cell properties, including malignancy. Moreover, gene manipulations against gangliosides have been shown to regulate cartilage metabolisms and chondrocyte differentiation in vivo and in vitro. This review highlights the potential of targeting gangliosides in the development of therapeutic strategies for osteoarthritis and cartilage injury and addresses promising directions for future research and treatment.
  • Articular cartilage corefucosylation regulates tissue resilience in osteoarthritis
    Kentaro Homan, Tomohiro Onodera, Hisatoshi Hanamatsu, Jun-ichi Furukawa, Daisuke Momma, Masatake Matsuoka, Norimasa Iwasaki
    eLife, 12, 24 Oct. 2023, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, This study aimed to investigate the glycan structural changes that occur before histological degeneration in osteoarthritis (OA) and to determine the mechanism by which these glycan conformational changes affect cartilage degeneration. An OA model was established in rabbits using mannosidase injection, which reduced high-mannose type N-glycans and led to cartilage degeneration. Further analysis of glycome in human OA cartilage identified specific corefucosylated N-glycan expression patterns. Inhibition of N-glycan corefucosylation in mice resulted in unrecoverable cartilage degeneration, while cartilage-specific blocking of corefucosylation led to accelerated development of aging-associated and instability-induced OA models. We conclude that α1,6 fucosyltransferase is required postnatally to prevent preosteoarthritic deterioration of articular cartilage. These findings provide a novel definition of early OA and identify glyco-phenotypes of OA cartilage, which may distinguish individuals at higher risk of progression.
  • A Review: Methodologies to Promote the Differentiation of Mesenchymal Stem Cells for the Regeneration of Intervertebral Disc Cells Following Intervertebral Disc Degeneration.
    Takashi Ohnishi, Kentaro Homan, Akira Fukushima, Daisuke Ukeba, Norimasa Iwasaki, Hideki Sudo
    Cells, 12, 17, 28 Aug. 2023, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Intervertebral disc (IVD) degeneration (IDD), a highly prevalent pathological condition worldwide, is widely associated with back pain. Treatments available compensate for the impaired function of the degenerated IVD but typically have incomplete resolutions because of their adverse complications. Therefore, fundamental regenerative treatments need exploration. Mesenchymal stem cell (MSC) therapy has been recognized as a mainstream research objective by the World Health Organization and was consequently studied by various research groups. Implanted MSCs exert anti-inflammatory, anti-apoptotic, and anti-pyroptotic effects and promote extracellular component production, as well as differentiation into IVD cells themselves. Hence, the ultimate goal of MSC therapy is to recover IVD cells and consequently regenerate the extracellular matrix of degenerated IVDs. Notably, in addition to MSC implantation, healthy nucleus pulposus (NP) cells (NPCs) have been implanted to regenerate NP, which is currently undergoing clinical trials. NPC-derived exosomes have been investigated for their ability to differentiate MSCs from NPC-like phenotypes. A stable and economical source of IVD cells may include allogeneic MSCs from the cell bank for differentiation into IVD cells. Therefore, multiple alternative therapeutic options should be considered if a refined protocol for the differentiation of MSCs into IVD cells is established. In this study, we comprehensively reviewed the molecules, scaffolds, and environmental factors that facilitate the differentiation of MSCs into IVD cells for regenerative therapies for IDD.
  • Simultaneous and sialic acid linkage-specific N- and O-linked glycan analysis by ester-to-amide derivatization.
    Hisatoshi Hanamatsu, Yoshiaki Miura, Takashi Nishikaze, Ikuko Yokota, Kentaro Homan, Tomohiro Onodera, Yoshihiro Hayakawa, Norimasa Iwasaki, Jun-Ichi Furukawa
    Glycoconjugate journal, 40, 2, 259, 267, Apr. 2023, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Characterization of O-glycans linked to serine or threonine residues in glycoproteins has mostly been achieved using chemical reaction approaches because there are no known O-glycan-specific endoglycosidases. Most O-glycans are modified with sialic acid residues at the non-reducing termini through various linkages. In this study, we developed a novel approach for sialic acid linkage-specific O-linked glycan analysis through lactone-driven ester-to-amide derivatization combined with non-reductive β-elimination in the presence of hydroxylamine. O-glycans released by non-reductive β-elimination were efficiently purified using glycoblotting via chemoselective ligation between carbohydrates and a hydrazide-functionalized polymer, followed by modification of methyl or ethyl ester groups of sialic acid residues on solid-phase. In-solution lactone-driven ester-to-amide derivatization of ethyl-esterified O-glycans was performed, and the resulting sialylated glycan isomers were discriminated by mass spectrometry. In combination with PNGase F digestion, we carried out simultaneous, quantitative, and sialic acid linkage-specific N- and O-linked glycan analyses of a model glycoprotein and human cartilage tissue. This novel glycomic approach will facilitate detailed characterization of biologically relevant sialylated N- and O-glycans on glycoproteins.
  • Establishment of the removal method of undifferentiated induced pluripotent stem cells coexisting with chondrocytes using R-17F antibody.
    Takuji Miyazaki, Hisatoshi Hanamatsu, Tomohiro Onodera, Jun-Ichi Furukawa, Liang Xu, Kentaro Homan, Rikiya Baba, Toshisuke Kawasaki, Norimasa Iwasaki
    Regenerative medicine, 17, 11, 793, 803, 26 Sep. 2022, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Aim: Tumorigenicity of residual undifferentiated induced pluripotent stem cells (iPSCs) is a major concern. The purpose of this study was to investigate the optimal conditions for removal of iPSCs using R-17F antibody, which recognizes specific glycosphingolipids glycans on undifferentiated iPSCs and exhibits selective cytotoxicity to iPSCs. Materials & methods: After adding of R-17F and secondary antibody to co-cultured iPSCs and chondrocytes, residual iPSCs were quantitatively evaluated by iPS specific glycome analysis. Results: Undifferentiated iPSCs were sufficiently removed using R-17F in combination with an equal amount of a secondary antibody. Furthermore, teratomas were not observed upon transplantation of co-cultured cells pretreated under the same conditions into testes of immunodeficient mice. Conclusion: This removal method incorporating R-17F may be useful for regenerative medicine using iPSCs.
  • Comprehensive validation of a wearable foot sensor system for estimating spatiotemporal gait parameters by simultaneous three-dimensional optical motion analysis.
    Kentaro Homan, Keizo Yamamoto, Ken Kadoya, Naoki Ishida, Norimasa Iwasaki
    BMC sports science, medicine & rehabilitation, 14, 1, 71, 71, 17 Apr. 2022, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, BACKGROUND: Use of a wearable gait analysis system (WGAS) is becoming common when conducting gait analysis studies due to its versatility. At the same time, its versatility raises a concern about its accuracy, because its calculations rely on assumptions embedded in its algorithms. The purpose of the present study was to validate twenty spatiotemporal gait parameters calculated by the WGAS by comparison with simultaneous measurements taken with an optical motion capture system (OMCS). METHODS: Ten young healthy volunteers wore two inertial sensors of the commercially available WGAS, Physilog®, on their feet and 23 markers for the OMCS on the lower part of the body. The participants performed at least three sets of 10-m walk tests at their self-paced speed in the laboratory equipped with 12 high-speed digital cameras with embedded force plates. To measure repeatability, all participants returned for a second day of testing within two weeks. RESULTS: Twenty gait parameters calculated by the WGAS had a significant correlation with the ones determined by the OMCS. Bland and Altman analysis showed that the between-device agreement for twenty gait parameters was within clinically acceptable limits. The validity of the gait parameters generated by the WGAS was found to be excellent except for two parameters, swing width and maximal heel clearance. The repeatability of the WGAS was excellent when measured between sessions. CONCLUSION: The present study showed that spatiotemporal gait parameters estimated by the WGAS were reasonably accurate and repeatable in healthy young adults, providing a scientific basis for applying this system to clinical studies.
  • Optical coherence tomography evaluation of the spatiotemporal effects of 3D bone marrow stromal cell culture using a bioreactor.
    Jun Yamaguchi, Tomohiro Onodera, Kentaro Homan, Xu Liang, Masatake Matsuoka, Takuji Miyazaki, Hosokawa Yoshiaki, Mitsuru Saito, Norimasa Iwasaki
    Journal of biomedical materials research. Part B, Applied biomaterials, 110, 8, 1853, 1861, 09 Mar. 2022, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Performing cell culture in a three-dimensional (3D) environment has various advantages. In cartilage tissue engineering, 3D in vitro cultures utilizing biomaterials and bioreactors can mimic the biological environment. However, the biggest drawback of these 3D culture systems is a limited ability to evaluate 3D cell distribution. Optical coherence tomography (OCT) has recently been used to evaluate 3D cellular morphology and structure in a timely manner. Here, we showed that OCT could be used to visually assess the distribution and the morphology of bone marrow stromal cells under chondrogenic 3D cultivation using alginate gels and rotary culture. In particular, OCT was able to visualize living cells embedded in alginate gels in a non-destructive and 3D manner, as well as quantitatively evaluate cell distribution and spheroid volume. We also found that cells were centralized in rotary culture but peripherally distributed in static culture, while rotary culture enhanced the hypertrophy of marrow stromal cells (MSCs) embedded in alginate gels. Together, our findings demonstrate that OCT can be used to evaluate the spatiotemporal effects of 3D cultivation using alginate gels and rotary culture. Therefore, this method may allow the observation of pre-cultured tissue over time and the optimization of culture conditions for regenerative tissue engineering.
  • Establishment of a New Qualitative Evaluation Method for Articular Cartilage by Dynamic T2w MRI Using a Novel Contrast Medium as a Water Tracer.
    Yoshiaki Hosokawa, Tomohiro Onodera, Kentaro Homan, Jun Yamaguchi, Kohsuke Kudo, Hiroyuki Kameda, Hiroyuki Sugimori, Norimasa Iwasaki
    Cartilage, 13, 3, 19476035221111503, 19476035221111503, 2022, [International Magazine]
    English, Scientific journal, OBJECTIVE: In the early stages of cartilage damage, diagnostic methods focusing on the mechanism of maintaining the hydrostatic pressure of cartilage are thought to be useful. 17O-labeled water, which is a stable isotope of oxygen, has the advantage of no radiation exposure or allergic reactions and can be detected by magnetic resonance imaging (MRI). This study aimed to evaluate MRI images using 17O-labeled water in a rabbit model. DESIGN: Contrast MRI with 17O-labeled water and macroscopic and histological evaluations were performed 4 and 8 weeks after anterior cruciate ligament transection surgery in rabbits. A total of 18 T2-weighted images were acquired, and 17O-labeled water was manually administered on the third scan. The 17O concentration in each phase was calculated from the signal intensity at the articular cartilage. Macroscopic and histological grades were evaluated and compared with the 17O concentration. RESULTS: An increase in 17O concentration in the macroscopic and histologically injured areas was observed by MRI. Macroscopic evaluation showed that the 17O concentration significantly increased in the damaged site group. Histological evaluations also showed that 17O concentrations significantly increased at 36 minutes 30 seconds after initiating MRI scanning in the Osteoarthritis Research Society International (OARSI) grade 3 (0.493 in grade 0, 0.659 in grade 1, 0.4651 in grade 2, and 0.9964 in grade 3, P < 0.05). CONCLUSION: 17O-labeled water could visualize earlier articular cartilage damage, which is difficult to detect by conventional methods.
  • Quantification of Cartilage Surface Degeneration by Curvature Analysis Using 3D Scanning in a Rabbit Model.
    Dawei Liang, Tomohiro Onodera, Masanari Hamasaki, Ryosuke Hishimura, Kentaro Homan, Liang Xu, Yuan Tian, Satoshi Kanai, Norimasa Iwasaki
    Cartilage, 13, 2_suppl, 19476035211059597, 19476035211059597, 20 Nov. 2021, [Peer-reviewed], [International Magazine]
    English, Scientific journal, OBJECTIVE: Accurate analysis to quantify cartilage morphology is critical for evaluating degenerative conditions in osteoarthritis (OA). Three-dimensional (3D) optical scanning provides 3D data for the entire cartilage surface; however, there is no consensus on how to quantify it. Our purpose was to validate a 3D method for evaluating spatiotemporal alterations in degenerative cartilages in a rabbit OA model by analyzing their curvatures at various stages of progression. DESIGN: Twelve rabbits underwent anterior cruciate ligament transection (ACLT) unilaterally and were divided into 4 groups: 4 weeks control, 4 weeks OA, 8 weeks control, and 8 weeks OA. 3D scanning, India ink staining, and histological assessments were performed in all groups. In 3D curvature visualization, the surfaces of the condyles were divided into 8 areas. The standard deviations (SD) of mean curvatures from all vertices of condylar surfaces and subareas were calculated. RESULTS: Regarding the site of OA change, curvature analysis was consistent with India ink scoring. The SD of mean curvature correlated strongly with the India ink Osteoarthritis Research Society International (OARSI) score. In curvature histograms, the curvature distribution in OA was more scattered than in control. Of the 8 areas, significant OA progression in the posterolateral part of the lateral condyle (L-PL) was observed at 4 weeks. The histology result was consistent with the 3D evaluation in terms of representative section. CONCLUSIONS: This study demonstrated that 3D scanning with curvature analysis can quantify the severity of cartilage degeneration objectively. Furthermore, the L-PL was found to be the initial area where OA degeneration occurred in the rabbit ACLT model.
  • Ultrapurified Alginate Gel Containing Bone Marrow Aspirate Concentrate Enhances Cartilage and Bone Regeneration on Osteochondral Defects in a Rabbit Model.
    Liang Xu, Atsushi Urita, Tomohiro Onodera, Ryosuke Hishimura, Takayuki Nonoyama, Masanari Hamasaki, Dawei Liang, Kentaro Homan, Jian Ping Gong, Norimasa Iwasaki
    The American journal of sports medicine, 49, 8, 2199, 2210, Jul. 2021, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: Ultrapurified alginate (UPAL) gel implantation has been demonstrated as effective in cartilage repair for osteochondral defects; however, cell transplantation within UPAL gels would be required to treat larger defects. HYPOTHESIS: The combination of UPAL gel and bone marrow aspirate concentrate (BMAC) would enhance cartilage repair and subchondral bone repair for large osteochondral defects. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 104 osteochondral defects (1 defect per knee) of 52 rabbits were randomly divided into 4 groups (26 defects per group): defects without any treatment (Defect group), defects treated using UPAL gel alone (UPAL group), defects treated using UPAL gel containing allogenic bone marrow mesenchymal stromal cells (UPAL-MSC group), and defects treated using UPAL gel containing BMAC (UPAL-BMAC group). At 4 and 16 weeks postoperatively, macroscopic and histologic evaluations and measurements of repaired subchondral bone volumes of reparative tissues were performed. Collagen orientation and mechanical properties of the reparative tissue were assessed at 16 weeks. RESULTS: The defects in the UPAL-BMAC group were repaired with hyaline-like cartilage with well-organized collagen structures. The histologic scores at 4 weeks were significantly higher in the UPAL-BMAC group (16.9 ± 2.0) than in the Defect group (4.7 ± 1.9; P < .05), the UPAL group (10.0 ± 3.3; P < .05), and the UPAL-MSC group (12.2 ± 2.9; P < .05). At 16 weeks, the score in the UPAL-BMAC group (24.4 ± 1.7) was significantly higher than those in the Defect group (9.0 ± 3.7; P < .05), the UPAL group (14.2 ± 3.9; P < .05), and the UPAL-MSC group (16.3 ± 3.6; P < .05). At 4 and 16 weeks, the macroscopic evaluations were significantly superior in the UPAL-BMAC group compared with the other groups, and the values of repaired subchondral bone volumes in the UPAL-BMAC group were significantly higher than those in the Defect and UPAL groups. The mechanical properties of the reparative tissues were significantly better in the UPAL-BMAC group than in the other groups. CONCLUSION: The implantation of UPAL gel containing BMAC-enhanced hyaline-like cartilage repair and subchondral bone repair of osteochondral defects in a rabbit knee model. CLINICAL RELEVANCE: These data support the potential clinical application of 1-step treatment for large osteochondral defects using biomaterial implantation with cell transplantation.
  • Flightless I is a catabolic factor of chondrocytes that promotes hypertrophy and cartilage degeneration in osteoarthritis.
    Taku Ebata, Mohamad Alaa Terkawi, Masanari Hamasaki, Gen Matsumae, Tomohiro Onodera, Mahmoud Khamis Aly, Shunichi Yokota, Hend Alhasan, Tomohiro Shimizu, Daisuke Takahashi, Kentaro Homan, Ken Kadoya, Norimasa Iwasaki
    iScience, 24, 6, 102643, 102643, 25 Jun. 2021, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Synovial macrophages that are activated by cartilage fragments initiate synovitis, a condition that promotes hypertrophic changes in chondrocytes leading to cartilage degeneration in OA. In this study, we analyzed the molecular response of chondrocytes under condition of this type of stimulation to identify a molecular therapeutic target. Stimulated macrophages promoted hypertrophic changes in chondrocytes resulting in production of matrix-degrading enzymes of cartilage. Among the top-upregulated genes, FliI was found to be released from activated chondrocytes and exerted autocrine/paracrine effects on chondrocytes leading to an increase in expression of catabolic and hypertrophic factors. Silencing FliI in stimulated cells significantly reduced expression of catabolic and hypertrophic factors in cocultured chondrocytes. Our further results demonstrated that the FliI-TLR4-ERK1/2 axis is involved in the hypertrophic signaling of chondrocytes and catabolism of cartilage. Our findings provide a new insight into the pathogenesis of OA and identify a potentially new molecular target for diagnostics and therapeutics.
  • Alterations of Glycosphingolipid Glycans and Chondrogenic Markers during Differentiation of Human Induced Pluripotent Stem Cells into Chondrocytes
    Liang Xu, Hisatoshi Hanamatsu, Kentaro Homan, Tomohiro Onodera, Takuji Miyazaki, Jun-ichi Furukawa, Kazutoshi Hontani, Yuan Tian, Rikiya Baba, Norimasa Iwasaki
    Biomolecules, 10, 12, Dec. 2020, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Due to the limited intrinsic healing potential of cartilage, injury to this tissue may lead to osteoarthritis. Human induced pluripotent stem cells (iPSCs), which can be differentiated into chondrocytes, are a promising source of cells for cartilage regenerative therapy. Currently, however, the methods for evaluating chondrogenic differentiation of iPSCs are very limited; the main techniques are based on the detection of chondrogenic genes and histological analysis of the extracellular matrix. The cell surface is coated with glycocalyx, a layer of glycoconjugates including glycosphingolipids (GSLs) and glycoproteins. The glycans in glycoconjugates play important roles in biological events, and their expression and structure vary widely depending on cell types and conditions. In this study, we performed a quantitative GSL-glycan analysis of human iPSCs, iPSC-derived mesenchymal stem cell like cells (iPS-MSC like cells), iPS-MSC-derived chondrocytes (iPS-MSC-CDs), bone marrow-derived mesenchymal stem cells (BMSCs), and BMSC-derived chondrocytes (BMSC-CDs) using glycoblotting technology. We found that GSL-glycan profiles differed among cell types, and that the GSL-glycome underwent a characteristic alteration during the process of chondrogenic differentiation. Furthermore, we analyzed the GSL-glycome of normal human cartilage and found that it was quite similar to that of iPS-MSC-CDs. This is the first study to evaluate GSL-glycan structures on human iPS-derived cartilaginous particles under micromass culture conditions and those of normal human cartilage. Our results indicate that GSL-glycome analysis is useful for evaluating target cell differentiation and can thus support safe regenerative medicine.
  • Which Contributes to Meniscal Repair, the Synovium or the Meniscus? An In Vivo Rabbit Model Study With the Freeze-Thaw Method.
    WooYoung Kim, Tomohiro Onodera, Eiji Kondo, Mohamad Alaa Terkawi, Kentaro Homan, Ryosuke Hishimura, Norimasa Iwasaki
    The American journal of sports medicine, 48, 6, 1406, 1415, May 2020, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: During meniscal tissue repair, the origin of the reparative cells of damaged meniscal tissue remains unclear. HYPOTHESIS: Comparison of the influence between meniscal and synovial tissues on meniscal repair by the in vivo freeze-thaw method would clarify the origin of meniscal reparative cells. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 48 mature Japanese white rabbits were divided into 4 groups according to the tissue (meniscal or synovial) that received freeze-thaw treatment. The meniscus of each group had a 2 mm-diameter cylindrical defect filled with alginate gel. Macroscopic and histologic evaluations of the reparative tissues were performed at 1, 3, and 6 weeks postoperatively. Additional postoperative measurements included cell density, which was the number of meniscal cells in the cut area per cut area (mm2) of meniscus; cell density ratio, which was the cell density of the sample from each group per the average cell density of the intact meniscus; and cell death rate, which was the number of cells stained by propidium iodide per the number of cells stained by Hoechst 33342 of the meniscal tissue adjacent to the defect. RESULTS: The macroscopic and histologic evaluations of the non-synovium freeze-thaw groups were significantly superior to those of the synovium freeze-thaw groups at 3 and 6 weeks postoperatively. Additionally, the meniscal cell density ratio and cell death rate in the freeze-thaw groups were significantly lower than those in the non-meniscal freeze-thaw groups at 3 and 6 weeks postoperatively. CONCLUSION: The freeze-thawed meniscus recovered few cells in its tissue even after 6 weeks. However, the defect was filled with fibrochondrocytes and proteoglycan when the synovium was intact. On the basis of these results, it is concluded that synovial cells are the primary contributors to meniscal injury repair. CLINICAL RELEVANCE: In meniscal tissue engineering, there is no consensus on the best cell source for meniscal repair. Based on this study, increasing the synovial activity and contribution should be the main objective of meniscal tissue engineering. This study can establish the foundation for future meniscal tissue engineering.
  • CCL21/CCR7 axis regulating juvenile cartilage repair can enhance cartilage healing in adults
    Kentaro Homan
    Scientific Reports, 9, 1, 5165, 5165, Dec. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Juvenile tissue healing is capable of extensive scarless healing that is distinct from the scar-forming process of the adult healing response. Although many growth factors can be found in the juvenile healing process, the molecular mechanisms of juvenile tissue healing are poorly understood. Here we show that juvenile mice deficient in the chemokine receptor CCR7 exhibit diminished large-scale healing potential, whereas CCR7-depleted adult mice undergo normal scar-forming healing similar to wild type mice. In addition, the CCR7 ligand CCL21 was transiently expressed around damaged cartilage in juvenile mice, whereas it is rarely expressed in adults. Notably, exogenous CCL21 administration to adults decreased scar-forming healing and enhanced hyaline-cartilage repair in rabbit osteochondral defects. Our data indicate that the CCL21/CCR7 axis may play a role in the molecular control mechanism of juvenile cartilage repair, raising the possibility that agents modulating the production of CCL21 in vivo can improve the quality of cartilage repair in adults. Such a strategy may prevent post-traumatic arthritis by mimicking the self-repair in juvenile individuals.
  • Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
    Takuji Miyazaki, Hisatoshi Hanamatsu, Liang Xu, Tomohiro Onodera, Jun-ichi Furukawa, Kentaro Homan, Rikiya Baba, Toshisuke Kawasaki, Norimasa Iwasaki
    International Journal of Molecular Sciences, 21, 1, Dec. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Cartilage damage may eventually lead to osteoarthritis because it is difficult to repair. Human-induced pluripotent stem cell (iPSC)-derived chondrocytes may potentially be used to treat cartilage damage, but the tumorigenicity of iPSCs is a major concern for their application in regenerative medicine. Many glycoconjugates serve as stem cell markers, and glycosphingolipids (GSLs) including H type 1 antigen (Fucα1-2Galβ1-3GlcNAc) have been expressed on the surface of iPSCs. The purpose of the present study was to investigate whether GSL-glycome analysis is useful for quality control of residual iPSCs in chondrocytes. We performed GSL-glycome analysis of undifferentiated iPSCs in chondrocytes by combining glycoblotting and aminolysis-sialic acid linkage-specific alkylamidation (SALSA) method, enabling the detection of small quantities of iPSC-specific GSL-glycans from 5 × 104 cells. Furthermore, we estimated the residual amount of iPSCs using R-17F antibody, which possesses cytotoxic activity toward iPSCs that is dependent on the Lacto-N-fucopentaose I (LNFP I) of GSL. Moreover, we could detect a small number of LNFP I during mesenchymal stem cells (MSCs) differentiation from iPSCs. This is the first demonstration that GSL-glycome analysis is useful for detecting undifferentiated iPSCs, and can thereby support safe regenerative medicine.
  • Alteration of the Total Cellular Glycome during Late Differentiation of Chondrocytes
    Kentaro Homan, Hisatoshi Hanamatsu, Jun-ichi Furukawa, Kazue Okada, Ikuko Yokota, Tomohiro Onodera, Norimasa Iwasaki
    International Journal of Molecular Sciences, 20, 14, 19 Jul. 2019, [Peer-reviewed], [Lead author], [International Magazine]
    English, Scientific journal, In normal articular cartilage, chondrocytes do not readily proliferate or terminally differentiate, and exhibit a low level of metabolism. Hypertrophy-like changes of chondrocytes have been proposed to play a role in the pathogenesis of osteoarthritis by inducing protease-mediated cartilage degradation and calcification; however, the molecular mechanisms underlying these changes are unclear. Glycans are located on the outermost cell surface. Dynamic cellular differentiation can be monitored and quantitatively characterized by profiling the glycan structures of total cellular glycoproteins. This study aimed to clarify the alterations in glycans upon late differentiation of chondrocytes, during which hypertrophy-like changes occur. Primary mouse chondrocytes were differentiated using an insulin-induced chondro-osteogenic differentiation model. Comprehensive glycomics, including N-glycans, O-glycans, free oligosaccharides, glycosaminoglycan, and glycosphingolipid, were analyzed for the chondrocytes after 0-, 10- and 20-days cultivation. The comparison and clustering of the alteration of glycans upon hypertrophy-like changes of primary chondrocytes were performed. Comprehensive glycomic analyses provided complementary alterations in the levels of various glycans derived from glycoconjugates during hypertrophic differentiation. In addition, expression of genes related to glycan biosynthesis and metabolic processes was significantly correlated with glycan alterations. Our results indicate that total cellular glycan alterations are closely associated with chondrocyte hypertrophy and help to describe the glycophenotype by chondrocytes and their hypertrophic differentiation. our results will assist the identification of diagnostic and differentiation biomarkers in the future.
  • Depletion of glycosphingolipids induces excessive response of chondrocytes under mechanical stress
    Kentaro Homan
    Journal of Biomechanics, 94, 22, 30, Jul. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, Glycosphingolipids (GSLs) are ubiquitous membrane components that play an indispensable role in maintaining chondrocyte homeostasis. To gain better insight into roles of GSLs, we studied the effects of GSL-deletion on the physiological responses of chondrocytes to mechanical stress. Mice lacking Ugcg gene (Ugcg-/-) were genetically generated to obtain GSL-deficient mice, and their chondrocytes from the joints were used for functional analyses in vitro culture experiments. The cells were seeded in a three-dimensional collagen gel and subjected to 5%, 10% or 16% cyclic tensile strain for either 3 or 24 h. The gene expressions of chondrocyte anabolic and catabolic factors, and the induction of Ca2+ signaling were analyzed. Our results revealed that chondrocytes derived from GSL-deficient mice exhibited an elevation in the expression of catabolic factors (ADAMTS-5, MMP-13) following the exposure to strain with amplitudes of 10%. Likewise, applying cyclic tensile strain with these amplitudes resulted in an increased Ca2+ oscillation ratio in chondrocytes from GSL-deficient as compared to the ratio from control mice. These results demonstrated that deletion of GSL stimulated the catabolic responses of chondrocytes to mechanical stress via the augmentation of the sensitivity to mechanical stress that may lead to the cartilage deterioration. These findings suggest that the regulation of the physiological responses of chondrocytes by GSLs could be a potential target in a therapeutic intervention in osteoarthritis.
  • Preoperative radiographic and clinical factors associated with postoperative floating of the lesser toes after resection arthroplasty for rheumatoid forefoot deformity
    Onodera, Tomohiro, Nakano, Hiroaki, Homan, Kentaro, Kondo, Eiji, Iwasaki, Norimasa
    BMC musculoskeletal disorders, 20, 1, 87, 87, Feb. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: This study aimed to clarify the characteristics associated with postoperative floating of the lesser toes, especially focusing on the medial and lateral lessor toes, after arthrodesis of the first metatarsophalangeal joint and resection arthroplasty of the lessor toes in rheumatoid forefoot deformity. METHODS: Fourty-seven feet of 43 people who underwent resection arthroplasty of the metatarsal head of the lesser toes for rheumatoid arthritis of the metatarsophalangeal joints were included. We retrospectively evaluated the preoperative radiographic findings and clinical characteristics of the patients, and the occurrence of postoperative floating of the lesser toes. The mean duration of follow-up was 36.5 (range 12 to 114) months. RESULTS: Preoperative dislocation grades of the second and third toes that demonstrated postoperative floating were significantly higher than those of toes that did not experience postoperative floating. The hallux valgus deformity before surgery was significantly more severe in toes with postoperative floating of the second and third lessor toes than those with no floating (p < 0.05). In addition, the Japanese Society for Surgery of the Foot (JSSF) hallux scale scores before surgery in toes with postoperative floating of the fourth and fifth lessor toes were significantly worse than those in non-dislocating toes (p < 0.05). CONCLUSIONS: The preoperative condition of the first metatarsophalangeal joint, including hallux valgus deformity, pain, range of motion, activity of daily living, and function is significantly different between postoperative floating of the lesser toes and non-floating of them after resection arthroplasty for rheumatoid forefoot deformity.
  • A Novel Cartilage Fragments Stimulation Model Revealed that Macrophage Inflammatory Response Causes an Upregulation of Catabolic Factors of Chondrocytes In Vitro
    Hamasaki, Masanari, Terkawi, Mohamad Alaa, Onodera, Tomohiro, Homan, Kentaro, Iwasaki, Norimasa
    Cartilage, 12, 3, 1947603519828426, 1947603519828426, {SAGE} Publications, Feb. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, OBJECTIVE: Osteoarthritis is a progressive joint disease characterized by cartilage degradation and synovial inflammation. Presence of cartilage fragments in the joint due to degradation of cartilage is thought to be associated with local inflammatory response and progressive osteoarthritic process. Understanding the mechanism by which cartilage fragments elicit this destructive process should aid in designing novel therapeutic approaches. Therefore, objective of current study is to establish an in vitro model to examine the cross-talk between chondrocytes and cartilage fragments-stimulated macrophages. DESIGN: Cartilage fragments were prepared from femoral head cartilages of mice and analyzed using a scanning electron microscope and particle size analyzer. Bone marrow-derived macrophages were co-cultured with cartilage fragments and chondrocytes using transwell co-culture system. Macrophage inflammatory mediators in supernatant of cultures were determined by ELISA and gene expression of macrophages and chondrocyte were quantified by qRT-PCR. RESULTS: Shapes of cartilage fragments were irregular with sizes ranged between 0.54 and 55 μm. Macrophages cultured with cartilage fragments released significantly higher concentrations of TNF-α, IL-6, and NO than those of mock and control. Consistently, gene expressions of TNF-α, IL-6, and MMP-9 were significantly increased in stimulated macrophages. The elevation in production of pro-inflammatory molecules in stimulated macrophages cultures were coincident with an increase in gene expression of chondrocyte MMP-13, iNOS, and IL-6. CONCLUSION: We developed an in vitro co-culture model to study the impact of stimulation of macrophage by cartilage fragments on the expression of chondrocyte carbolic factors. Our results revealed that cartilage fragments triggered macrophages inflammatory response that enhanced the production of chondrocyte catabolic factors.
  • Osteochondral Autograft Transplantation Technique Augmented by an Ultrapurified Alginate Gel Enhances Osteochondral Repair in a Rabbit Model
    Hishimura, Ryosuke, Onodera, Tomohiro, Hontani, Kazutoshi, Baba, Rikiya, Homan, Kentaro, Matsubara, Shinji, Joutoku, Zenta, Kim, WooYoung, Nonoyama, Takayuki, Kurokawa, Takayuki, Gong, Jian Ping, Iwasaki, Norimasa
    The American Journal of Sports Medicine, 47, 2, 363546518817527, 363546518817527, Jan. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: One of the most important limitations of osteochondral autograft transplantation (OAT) is the adverse effect on donor sites in the knee. To decrease the number and/or size of osteochondral defects, we devised a method with biomaterial implantation after OAT. HYPOTHESIS: OAT augmented by ultrapurified alginate (UPAL) gel enhances cartilage repair capacity. STUDY DESIGN: Controlled laboratory study. METHODS: Seventy-five osteochondral defects in rabbits were divided into 3 groups: osteochondral defects with OAT alone, defects with OAT augmented by UPAL gel (combined group), and defects without intervention as controls. Macroscopic and histological evaluations of the reparative tissues were performed at 4 and 12 weeks postoperatively. Histological evaluation of graft cartilage degradation was also performed. To evaluate the effects of UPAL gel on graft healing, repaired bone volumes and osseointegration of the graft were evaluated. Collagen orientation and the mechanical properties of the reparative tissue and graft cartilage were also evaluated qualitatively. RESULTS: The macroscopic and histological evaluations of the combined group were significantly superior to the other groups at 12 weeks postoperatively. Regarding degenerative change of the graft, the histological scores of the combined group were significantly higher than those of the OAT-alone group. The values of repaired subchondral bone volumes and osseointegration of the graft were almost identical in both groups. Collagen orientation and the mechanical properties of the reparative tissue and graft cartilage were significantly better in the combined group than in the other groups. CONCLUSION: Administration of UPAL gel in OAT enhanced cartilage repair and protected graft cartilage without inhibiting subchondral bone repair and graft survival. CLINICAL RELEVANCE: OAT augmented by UPAL gel decreases the number and/or size of osteochondral grafts, minimizing the risk of donor site morbidity. This combination technique has the potential to improve clinical outcomes and expand the surgical indications for OAT.
  • Chondrogenic differentiation of mouse induced pluripotent stem cells (iPSCs) using the three-dimensional culture with ultra-purified alginate gel (UPAL gel)
    Hontani, Kazutoshi, Onodera, Tomohiro, Terashima, Michiyo, Momma, Daisuke, Matsuoka, Masatake, Baba, Rikiya, Joutoku, Zenta, Matsubara, Shinji, Homan, Kentaro, Hishimura, Ryosuke, Xu, Liang, Iwasaki, Norimasa
    Journal of Biomedical Materials Research. Part A, 107, 5, 1086, 1093, Jan. 2019, [Peer-reviewed], [International Magazine]
    English, Scientific journal, As articular cartilages have rarely healed by themselves because of their characteristics of avascularity and low cell density, surgical intervention is ideal for patients with cartilaginous injuries. Because of structural characteristics of the cartilage tissue, a three-dimensional culture of stem cells in biomaterials is a favorable system on cartilage tissue engineering. Induced pluripotent stem cells (iPSCs) are a new cell source in cartilage tissue engineering for its characteristics of self-renewal capability and pluripotency. However, the optimal cultivation condition for chondrogenesis of iPSCs is still unknown. Here we show that a novel chondrogenic differentiation method of iPSCs using the combination of three-dimensional cultivation in ultra-purified alginate gel (UPAL gel) and multi-step differentiation via mesenchymal stem cell-like cells (iPS-MSCs) could efficiently and specifically differentiate iPSCs into chondrocytes. The iPS-MSCs in UPAL gel culture sequentially enhanced the expression of chondrogenic marker without the upregulation of that of osteogenic and adipogenic marker and histologically showed homogeneous chondrogenic extracellular matrix formation. Our results suggest that the pluripotency of iPSCs can be controlled when iPSCs are differentiated into iPS-MSCs before embedding in UPAL gel. These results lead to the establishment of an efficient three-dimensional system to engineer artificial cartilage tissue from iPSCs for cartilage regeneration. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1086-1093, 2019.
  • Effects of Ultra-Purified Alginate Gel Implantation on Meniscal Defects in Rabbits
    Kim, WooYoung, Onodera, Tomohiro, Kondo, Eiji, Kawaguchi, Yasuyuki, Terkawi, Mohamad Alaa, Baba, Rikiya, Hontani, Kazutoshi, Joutoku, Zenta, Matsubara, Shinji, Homan, Kentaro, Hishimura, Ryosuke, Iwasaki, Norimasa
    The American Journal of Sports Medicine, 47, 3, 363546518816690, 363546518816690, Dec. 2018, [Peer-reviewed], [International Magazine]
    English, Scientific journal, BACKGROUND: Many tissue-engineered methods for meniscal repair have been studied, but their utility remains unclear. HYPOTHESIS: Implantation of low-endotoxin, ultra-purified alginate (UPAL) gel without cells could induce fibrocartilage regeneration on meniscal defects in rabbits. STUDY DESIGN: Controlled laboratory study. METHODS: Forty-two mature Japanese White rabbits were divided into 2 groups of 21 animals each. In each animal, a cylindrical defect measuring 2 mm in diameter was created with a biopsy punch on the anterior horn of the medial meniscus. In the control group, no treatment was applied on the left medial meniscal defect. In the UPAL gel group, the right medial meniscal defect was injected with the UPAL gel and gelated by a CaCl2 solution. Samples were evaluated at 3, 6, and 12 weeks postoperatively. For biomechanical evaluation, 6 additional samples from intact animals were used for comparison. RESULTS: The macroscopic score was significantly greater in the UPAL gel group than in the control group at 3 weeks (mean ± SE: 5.6 ± 0.82 vs 3.4 ± 0.83, P = .010), 6 weeks (5.9 ± 0.72 vs 2.5 ± 0.75, P = .026), and 12 weeks (5.2 ± 1.21 vs 1.0 ± 0.63, P = .020). The histological score was significantly greater in the UPAL group than in the control group at 3 weeks (2.1 ± 0.31 vs 1.2 ± 0.25, P = .029) and 12 weeks (2.2 ± 0.55 vs 0.3 ± 0.21, P = .016). The mean stiffness of the reparative tissue in the UPAL gel group was significantly greater than that in the control group at 6 weeks (24.325 ± 3.920 N/mm vs 8.723 ± 1.190 N/mm, P = .006) and at 12 weeks (27.804 ± 6.169 N/mm vs not applicable [because of rupture]). CONCLUSION: The UPAL gel enhanced the spontaneous repair of fibrocartilage tissues in a cylindrical meniscal defect in rabbits. CLINICAL RELEVANCE: These results imply that the acellular UPAL gel may improve the repair of traumatic meniscal injuries.
  • Coordinated existence of multiple gangliosides is required for cartilage metabolism
    Momma, Daisuke, Onodera, Tomohiro, Homan, Kentaro, Matsubara, Shinji, Sasazawa, Fumio, Furukawa, Junichi, Matsuoka, Masatake, Yamashita, Tadashi, Iwasaki, Norimasa
    Osteoarthritis and Cartilage, Nov. 2018, [Peer-reviewed], [International Magazine]
    Scientific journal
  • Bone Marrow Stimulation Technique Augmented by an Ultrapurified Alginate Gel Enhances Cartilage Repair in a Canine Model
    Baba, Rikiya, Onodera, Tomohiro, Matsuoka, Masatake, Hontani, Kazutoshi, Joutoku, Zenta, Matsubara, Shinji, Homan, Kentaro, Iwasaki, Norimasa
    The American Journal of Sports Medicine, 46, 8, 363546518770436, 363546518770436, May 2018, [Peer-reviewed], [International Magazine]
    English, Scientific journal
  • Depletion of Gangliosides Enhances Articular Cartilage Repair in Mice
    Matsuoka, Masatake, Onodera, Tomohiro, Homan, Kentaro, Sasazawa, Fumio, Furukawa, Jun-Ichi, Momma, Daisuke, Baba, Rikiya, Hontani, Kazutoshi, Joutoku, Zenta, Matsubara, Shinji, Yamashita, Tadashi, Iwasaki, Norimasa
    Scientific Reports, 7, 43729, 43729, Mar. 2017, [Peer-reviewed], [International Magazine]
    English, Scientific journal
  • The effect of changing toe direction on knee kinematics during drop vertical jump: a possible risk factor for anterior cruciate ligament injury
    Ishida, Tomoya, Yamanaka, Masanori, Takeda, Naoki, Homan, Kentaro, Koshino, Yuta, Kobayashi, Takumi, Matsumoto, Hisashi, Aoki, Yoshimitsu
    Knee surgery, sports traumatology, arthroscopy: official journal of the ESSKA, 23, 4, 1004, 1009, Apr. 2015, [Peer-reviewed], [International Magazine]
    English, Scientific journal
  • The measure of navicular height-Compared with measurement obtained from radiograph-
    Homan K, Yamanaka M, Okuyama A, Kudo K, Kida T, Kawashima N, Suzuki Y, Horiuchi H, Nakayama H
    Journal of Hokkaido Physical Therapy, 20, 13, 16, (公社)北海道理学療法士会, Apr. 2003, [Peer-reviewed], [Lead author], [Domestic magazines]
    Japanese
  • The Effect of Medial Arch Support Tape. Examination of the Healthy Volunteers
    Suzuki Y, Yamanaka M, Kida A, Matsumoto H, Horiuchi H, Nakayama H, Okuyama A, Kudo K, Homan K
    Journal of Hokkaido Physical Therapy, 19, 30, 33, (公社)北海道理学療法士会, Apr. 2002, [Peer-reviewed], [Domestic magazines]
    Japanese

Other Activities and Achievements

Books and other publications

Affiliated academic society

  • Apr. 2021 - Present
    日本運動器理学療法学会　　　　　　　　　　　　　　　
  • THE JAPANESE SOCIETY OF CARBOHYDRATE RESEARCH　　　　　　　　　　　　　　　
  • 臨床歩行分析研究会　　　　　　　　　　　　　　　
  • JAPANESE PHYSICAL THERAPY ASSOCIATION　　　　　　　　　　　　　　　
  • HOKKAIDO SOCIETY OF ORTHOPAEDIC AND TRAUMATOLOGY　　　　　　　　　　　　　　　
  • The Japanese Society of Cartilage Metabolism　　　　　　　　　　　　　　　
  • Orthopaedic Research Society　　　　　　　　　　　　　　　
  • Orthopedics Biomaterial Bureau　　　　　　　　　　　　　　　
  • THE JAPANESE SOCIETY FOR BIOMATERIALS　　　　　　　　　　　　　　　

Research Themes

  • Developing a gait-based system to detect early knee osteoarthritis.
    Grants-in-Aid for Scientific Research
    01 Apr. 2023 - 31 Mar. 2026
    宝満 健太郎
    Japan Society for the Promotion of Science, Grant-in-Aid for Early-Career Scientists, Hokkaido University, 23K16633
  • Chondrocyte glycome analysis reveals the pathogenesis of osteoarthritis
    Grants-in-Aid for Scientific Research Grant-in-Aid for Early-Career Scientists
    01 Apr. 2019 - 31 Mar. 2022
    Homan Kentaro
    Degeneration of cartilage, the main component of osteoarthritis (OA) lesions, is believed to begin with the destruction of glycan structures in proteoglycans. Although many studies on the pathogenesis of OA have focused on the hypertrophic differentiation of chondrocytes, none have addressed changes in glycoconjugate sugar chains. We have shown that N-glycans are altered before the appearance of histological changes in the cartilage matrix and that glycosphingolipids are involved in chondrocyte hypertrophy and OA progression. Therefore, in order to detect homologous glycan variation during cartilage tissue degeneration and chondrocyte differentiation, we succeeded in identifying glycans that play a important role in the onset of OA by performing qualitative and quantitative comparative analysis of glycan structural changes in all classes of glycans without limiting to one glycan class.
    Japan Society for the Promotion of Science, Grant-in-Aid for Early-Career Scientists, Hokkaido University, 19K18516
  • 培養細胞上の糖鎖抗原変化と自家細胞移植における免疫応答発生機序の解明
    Grants-in-Aid for Scientific Research Challenging Research (Exploratory)
    28 Jun. 2019 - 31 Mar. 2021
    岩崎 倫政, テルカウィ アラー, 古川 潤一, 宝満 健太郎
    2019年度は、１、継代培養前後のマウス軟骨細胞およびマウス骨髄間葉系幹細胞（BMSC）上の糖鎖構造変化が生じることを確認すること、２、継代培養による免疫応答変化を捉えるためのモデルの確立の２点を研究目標とし、研究を実施した。
    １、継代培養に伴う糖鎖構造変化の解析
    軟骨細胞は哺乳5日目のC57BL/6Nマウスの関節軟骨からコラゲナーゼDで処理して単離した。BMSCは、2週齢の同系マウスの大腿骨と脛骨を採取し、骨髄を長骨から洗い流し、骨片をコラゲナーゼIIで消化、放出された細胞を除去して消化された骨片を培養し、そこから移動して成長する線維芽細胞様細胞をBMSCとして継代培養を行った。各代の細胞上の糖鎖構造をグライコブロッティング法により網羅的に解析し、その構造推定を行った。
    ２、継代培養による免疫応答変化を捉えるためのモデル
    自家および同種細胞をそれぞれ平面培養にて3継代行い、各培養細胞を免疫細胞（マクロファージ）と共培養することにより軟骨細胞およびBMSCの賦活化を評価した。まずはじめにC57BL/6Nマウスの腹腔内マクロファージを単離し、細胞数を2.0×10^5 cellsに調整した。次に各継代数（passage3, 4, 6, 8）のBMSCを2.0×10^5 cellsに同数で調整してマクロファージとtranswellで共培養を行った。培地を回収し、免疫原性の評価としてELISA、TNF-α、IL-6などの産生を計測した。
    Japan Society for the Promotion of Science, Challenging Research (Exploratory), Hokkaido University, 19K22672
  • Functional analysis of cartilage glycans for the elucidation of osteoarthritis pathomechanisms
    Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    01 Apr. 2016 - 31 Mar. 2018
    Iwasaki Norimasa, HOMAN Kentaro
    Our hypothesis was that early osteoarthritic cahnges induced by alterations in cartilage glycans would be reversible. To test this hypothesis, we created in vivo and in vitro early osteoarthritic models based on alpha-mannosidase administration. The administration of alpha-mannosidase induced early osteoarthritis in in vivo and in vitro models. Additionally, this histological changes were recovered by the discontinuation of alpha-mannosidase adminiatrations. Future studies will be performed to clarify this recovery mechanism in early osteoarthritis pathogenesis.
    Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 16K15650

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