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Changes in the intra- and peri-cellular sclerostin distribution in lacuno-canalicular system induced by mechanical unloading

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A Correction to this article was published on 06 October 2020

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Abstract

Introduction

Mechanical stimuli regulate Sclerostin (Scl), a negative regulator of bone formation, expression in osteocytes. However, the detailed Scl distribution in osteocytes in response to mechanical unloading remains unclear.

Materials and methods

Twelve-week-old male rats were used. The sciatic and femoral nerves on the right side were excised as mechanical unloading treatment. A sham operation was performed on the left side. One week after neurotrauma, the bone density of the femora was evaluated by peripheral quantitative computed tomography, and immunofluorescence was performed in coronal sections of the femoral diaphysis. The mean fluorescence intensity and fluorescent profile of Scl from the marrow to the periosteal side were analyzed to estimate the Scl expression and determine to which side (marrow or periosteal) the Scl prefers to distribute in response to mechanical unloading. The most sensitive region indicated by the immunofluorescence results was further investigated by transmission electron microscopy (TEM) with immunogold staining to show the Scl expression changes in different subcellular structures.

Results

In femur distal metaphysis, neurotrauma-induced mechanical unloading significantly decreased the bone density, made the distribution of Scl closer to the marrow on the anterior and medial side, and increased the Scl expression only on the lateral side. TEM findings showed that only the expression of Scl in canaliculi was increased by mechanical unloading.

Conclusions

Our results showed that even short-term mechanical unloading is enough to decrease bone density, and mechanical unloading not only regulated the Scl expression but also changed the Scl distribution in both the osteocyte network and subcellular structures.

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Acknowledgements

The authors would like to thank Masumi Furutani and Megumi Tsukano, Central Research Laboratory, Okayama University Medical School, for their technical assistance in this study. This work was supported by a Grant-in-Aid for Scientific Research (to T. Iimura [18H02983], Y. Ishihara [17H04413], Z. Wang [19J11906] and H. Kamioka [16H05549, 19H03859]) from the Japan Society for the Promotion of Science, Japan.

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Author notes

  1. Ryuta Osumi and Ziyi Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Orthodontics, Okayama University Hospital, Okayama, Japan

    Ryuta Osumi, Yoshihito Ishihara & Naoya Odagaki

  2. Department of Orthodontics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Okayama, Okayama, 700-8558, Japan

    Ziyi Wang & Hiroshi Kamioka

  3. Research Fellow of Japan Society for the Promotion of Science, Tokyo, Japan

    Ziyi Wang

  4. Department of Pharmacology, Graduate School of Dental Medicine, Hokkaido University, Hokkaido, Japan

    Tadahiro Iimura

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  1. Ryuta Osumi
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Contributions

H.K., R.O., T.I., and Z.W. designed the study. R.O., N.O., T.I., and Y.I. conducted the study. Z.W. tested the existing code components, the programmed script for data processing, and drew the sketch. R.O. and Z.W. processed, analyzed, and visualized the data and wrote the manuscript. R.O., Z.W., T.I., Y.I., and H.K. interpreted the data and approved the final version of the manuscript. H.K. is responsible for the integrity of the data analysis. R.O and Z.W. contributed equally to this work.

Corresponding author

Correspondence to Hiroshi Kamioka.

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The original version of this article was revised due to the left panel of figure 3 was published incorrectly and corrected in this version.

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Online Resource 1. A video to show that the entire target leg was fully immobilized. In this video, the rat was walking with a slow dragging motion without lifting the right foot. We shot this video one day after the excision of the sciatic and femoral nerves. (MPG 10486 kb)

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Osumi, R., Wang, Z., Ishihara, Y. et al. Changes in the intra- and peri-cellular sclerostin distribution in lacuno-canalicular system induced by mechanical unloading. J Bone Miner Metab 39, 148–159 (2021). https://doi.org/10.1007/s00774-020-01135-9

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  • DOI: https://doi.org/10.1007/s00774-020-01135-9

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