Establishment of novel animal model of ectopic bone formation

• Project/Area Number: 26861208
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Orthopaedic surgery
• Research Institution: Kyushu Dental College (2015); Saitama Medical University (2014)
• Principal Investigator: Osawa Kenji 九州歯科大学, 歯学部, 助教 (70638238)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 骨形成 / 骨形成因子(BMP) / 超音波遺伝子導入 / 骨誘導 / 遺伝子導入 / ソノポレーション

Outline of Final Research Achievements

Bone morphogenetic proteins (BMPs) induce heterotypic bone formation in soft tissues, such as skeletal muscle, and induce osteoblastic differentiation of myoblasts in vitro. We attempted to establish an in vivo experimental model for heterotypic bone formation using ultrasound-mediated gene transfer (sonoporation). A mixture of BMP-encoding plasmidvector and a micro bubble echo-contrast agent was injected to the gastrocnemius muscle of mice, and ultrasound irradiation was performed at the injection site. Radiopacity was observed in the BMP transfection site and heterotypic endochondral ossification was observed in the muscle tissue, indicating that local expression of BMPcould induce the heterotypic ossification.Moreover, bone formation could enhance by cardiotoxin (CTX) injection before sonoporation. This model may be a useful tool to clarify mechanisms of BMP-induced bone formation in vivo.

Research Products

• [Journal Article] Comparison of human mesenchymal stem cells derived from bone marrow, synovial fluid, adult dental pulp, and exfoliated deciduous tooth pulp. (2016)
  • Author(s): Isobe Y, Koyama N, Nakao K, Osawa K, Ikeno M, Yamanaka S, Okubo Y, Fujimura K, Bessho K.
  • Journal Title: Int J Oral Maxillofac Surg. Volume: 45 Issue: 1 Pages: 124-131
  • DOI: 10.1016/j.ijom.2015.06.022
  • Peer Reviewed
• [Journal Article] The local CNP/GC-B system in the growth plate is responsible for physiological endochondral bone growth. (2015)
  • Author(s): Nakao, K., Osawa, K., Yasoda, A., Yamanaka, S., Fujii, T., Kondo, E., Koyama, N., Kanamoto, N., Miura, M., Kuwahara, K., Akiyama, H., Bessho, K., and Nakao, K..
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 1-11
  • DOI: 10.1038/srep10554
  • NAID: 120007100129
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Mutant activin-like kinase 2 in fibrodysplasia ossificans progressiva are activated via T203 by BMP type II receptor (2015)
  • Author(s): Fujimoto M, Ohte S, Osawa K, Miyamoto A, Tsukamoto S, Mizuta T, Kokabu S, Suda N, Katagiri T
  • Journal Title: Mol Endocrinol Volume: 29 Issue: 1 Pages: 140-152
  • DOI: 10.1210/me.2014-1301
  • Peer Reviewed / Open Access
• [Journal Article] Smad9 is a new type of transcriptional regulator in bone morphogenetic protein signaling. (2014)
  • Author(s): Tsukamoto S, Mizuta T, Fujimoto M, Ohte S, Osawa K, Miyamoto A, Yoneyama K, Murata E, Machiya A, Jimi E, Kokabu S, Katagiri T.
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 7596-7596
  • DOI: 10.1038/srep07596
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Establishment of a novel model of chondrogenesis using murine embryonic stem cells carrying fibrodysplasia ossificans progressiva-associated mutant ALK2. (2014)
  • Author(s): Fujimoto M, Ohte S, Shin M, Yoneyama K, Osawa K, Miyamoto A, Tsukamoto S, Mizuta T, Kokabu S, Machiya A, Okuda A, Suda N, Katagiri T.
  • Journal Title: Biochmical and Biophysical Research Communications Volume: 455 Issue: 3-4 Pages: 347-352
  • DOI: 10.1016/j.bbrc.2014.11.012
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The novel IκB kinase β inhibitor IMD-0560 prevents bone invasion by oral squamous cell carcinoma. (2014)
  • Author(s): 4.Tada Y, Kokabu S, Sugiyama G, Nakatomi C, Aoki K, Fukushima H, Osawa K, Sugamori Y, Ohya K, Okamoto M, Fujikawa T, Itai A, Matsuo K, Watanabe S, Jimi E.
  • Journal Title: Oncotarget Volume: 15 Pages: 12317-12330
  • Peer Reviewed
• [Presentation] Establishment of a new in vivo experimental model for heterotopic bone formation in skeletal muscle. (2014)
  • Author(s): Osawa K, Tsukamoto S, Fujimoto M, Miyamoto A, Mizta T, Katagiri T
  • Organizer: The 12th RCGM International Symposium of Academic Frontier
  • Place of Presentation: 埼玉医科大学３０周年記念講堂 (埼玉県日高市)
  • Year and Date: 2014-10-31 – 2014-11-01
• [Book] A Tissue Regeneration Approach to Bone and Cartilage Repair (2014)
  • Author(s): Katagiri T, Tsukamoto S, Osawa K, Kokabu, S.
  • Total Pages: 15
  • Publisher: Springer International Publishing
• [Book] 骨代謝 つくり，壊し，変える−そのメカニズムと最新治療 (2014)
  • Author(s): 片桐岳信、塚本翔、大澤賢次
  • Total Pages: 6
  • Publisher: 羊土社
• [Patent(Industrial Property Rights)] 抗ALK2抗体 (2016)
  • Inventor(s): 片桐岳信
  • Industrial Property Rights Holder: 大澤賢次、塚本翔
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-01-29