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Collagen production of osteoblasts revealed by ultra-high voltage electron microscopy

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

In the bone, collagen fibrils form a lamellar structure called the “twisted plywood-like model.” Because of this unique structure, bone can withstand various mechanical stresses. However, the formation of this structure has not been elucidated because of the difficulty of observing the collagen fibril production of the osteoblasts via currently available methods. This is because the formation occurs in the very limited space between the osteoblast layer and bone matrix. In this study, we used ultra-high-voltage electron microscopy (UHVEM) to observe collagen fibril production three-dimensionally. UHVEM has 3-MV acceleration voltage and enables us to use thicker sections. We observed collagen fibrils that were beneath the cell membrane of osteoblasts elongated to the outside of the cell. We also observed that osteoblasts produced collagen fibrils with polarity. By using AVIZO software, we observed collagen fibrils produced by osteoblasts along the contour of the osteoblasts toward the bone matrix area. Immediately after being released from the cell, the fibrils run randomly and sparsely. But as they recede from the osteoblast, the fibrils began to run parallel to the definite direction and became thick, and we observed a periodical stripe at that area. Furthermore, we also observed membrane structures wrapped around filamentous structures inside the osteoblasts. The filamentous structures had densities similar to the collagen fibrils and a columnar form and diameter. Our results suggested that collagen fibrils run parallel and thickly, which may be related to the lateral movement of the osteoblasts. UHVEM is a powerful tool for observing collagen fibril production.

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Acknowledgments

The authors would like to thank Toshiaki Hasegawa, Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, for technical assistance in this study; Naoko Kajimura, visiting research scholar of Japan Electron Optics Laboratory; Noriyuki Nagaoka, Laboratory for Electrons, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences; and Masaru Kaku, Division of Bioprosthodontics, Niigata University Graduate School of Medical and Dental Sciences, for his fruitful discussion. This work was supported by the Japan Society for the Promotion of Science in the form of Grants-in-Aid for Scientific Research (no. 25293419). The work at the research center for Ultra-High-Voltage Electron Microscopy, Osaka University (Handai Multifunctional Nano-Foundary) was supported by the Nano-technology Network Project of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

Conflict of interest

All authors have no conflicts of interest.

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Authors and Affiliations

  1. Department of Orthodontics, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikata, Kita-ku, Okayama, Okayama, 700-8525, Japan

    Rumiko Hosaki-Takamiya, Mana Hashimoto, Yuichi Imai, Tomoyo Tanaka, Noriaki Kawanabe & Hiroshi Kamioka

  2. Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, 7-1, Mihogaoka, Ibaraki, Osaka, 567-0047, Japan

    Tomoki Nishida, Naoko Yamada & Hirotaro Mori

  3. Department of Orthodontics and Dentofacial Orthopedics, Graduate School of Dentistry, Osaka University, 1-8 Yamada-Oka, Suita, Osaka, 565-0871, Japan

    Takashi Yamashiro

Authors
  1. Rumiko Hosaki-Takamiya
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  2. Mana Hashimoto
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  3. Yuichi Imai
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  4. Tomoki Nishida
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  5. Naoko Yamada
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  6. Hirotaro Mori
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  7. Tomoyo Tanaka
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  8. Noriaki Kawanabe
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  9. Takashi Yamashiro
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  10. Hiroshi Kamioka
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Corresponding author

Correspondence to Hiroshi Kamioka.

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Hosaki-Takamiya, R., Hashimoto, M., Imai, Y. et al. Collagen production of osteoblasts revealed by ultra-high voltage electron microscopy. J Bone Miner Metab 34, 491–499 (2016). https://doi.org/10.1007/s00774-015-0692-0

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  • DOI: https://doi.org/10.1007/s00774-015-0692-0

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