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Identification of a progenitor cell population destined to form fracture fibrocartilage callus in Dickkopf-related protein 3–green fluorescent protein reporter mice

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Fracture healing is a complex biological process involving the proliferation of mesenchymal progenitor cells, and chondrogenic, osteogenic, and angiogenic differentiation. The mechanisms underlying the proliferation and differentiation of mesenchymal progenitor cells remain unclear. Here, we demonstrate Dickkopf-related protein 3 (Dkk3) expression in periosteal cells using Dkk3–green fluorescent protein reporter mice. We found that proliferation of mesenchymal progenitor cells began in the periosteum, involving Dkk3-positive cell proliferation near the fracture site. In addition, Dkk3 was expressed in fibrocartilage cells together with smooth muscle α-actin and Col3.6 in the early phase of fracture healing as a cell marker of fibrocartilage cells. Dkk3 was not expressed in mature chondrogenic cells or osteogenic cells. Transient expression of Dkk3 disappeared in the late phase of fracture healing, except in the superficial periosteal area of fracture callus. The Dkk3 expression pattern differed in newly formed type IV collagen positive blood vessels and the related avascular tissue. This is the first report that shows Dkk3 expression in the periosteum at a resting state and in fibrocartilage cells during the fracture healing process, which was associated with smooth muscle α-actin and Col3.6 expression in mesenchymal progenitor cells. These fluorescent mesenchymal lineage cells may be useful for future studies to better understand fracture healing.

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Acknowledgments

This study was supported by JSPS KAKENHI (25870039), the Department of Defense (DAMD W81XWH07-2-0085), and the National Institutes of Health (AR052374).

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Authors and Affiliations

  1. Center for Regenerative Medicine and Skeletal Biology, Department of Reconstructive Sciences, School of Dental Medicine, University of Connecticut Health Center, Farmington, CT, USA

    Yu Mori, Douglas Adams, Yusuke Hagiwara, Ryu Yoshida & David W. Rowe

  2. Department of Orthopaedic Surgery, Graduate School of Tohoku University, 1-1 Seiryomachi, Aobaku, Sendai, Miyagi, 980-8574, Japan

    Yu Mori, Masayuki Kamimura & Eiji Itoi

Authors
  1. Yu Mori
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  2. Douglas Adams
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  3. Yusuke Hagiwara
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  4. Ryu Yoshida
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  5. Masayuki Kamimura
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  6. Eiji Itoi
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  7. David W. Rowe
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Correspondence to Yu Mori.

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The authors declare that they have no conflict of interest.

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774_2015_711_MOESM1_ESM.tif

Fig. S1 Fibrocartilage proliferation and Dkk3 green expression at day 7 after fracture. Left picture shows an image of safranin-O and hematoxylin staining, mature cartilage cells are positive for safranin-O red (arrow) (×10 magnification). Right picture shows Dkk3-green GFP image, Dkk3-green is not positive on the mature fibrocartilage cells, is expressed on the superficial area of mature fibrocartilage cells (arrow head) (×10 magnification). (TIFF 3417 kb)

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Mori, Y., Adams, D., Hagiwara, Y. et al. Identification of a progenitor cell population destined to form fracture fibrocartilage callus in Dickkopf-related protein 3–green fluorescent protein reporter mice. J Bone Miner Metab 34, 606–614 (2016). https://doi.org/10.1007/s00774-015-0711-1

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  • DOI: https://doi.org/10.1007/s00774-015-0711-1

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