Interleukin-6 Inhibitor Suppresses Hyperalgesia Without Improvement in Osteoporosis in a Mouse Pain Model of Osteoporosis

  • Hiroki WakabayashiEmail author
  • Sho Kato
  • Nobuto Nagao
  • Gaku Miyamura
  • Yohei Naito
  • Akihiro Sudo
Original Research


The aim of this study was to evaluate skeletal pain associated with osteoporosis and examine the inhibitory effect of interleukin-6 (IL-6) on pain in ovariectomized (OVX) mice. The mechanism of osteoporotic pain in OVX mice was evaluated by examining pain-related behavior and immunohistochemistry. The effects of IL-6 receptor inhibitor (IL-6i) on these parameters were also assessed. Eight-week-old female ddY mice were ovariectomized and assigned to three groups: OVX mice treated with vehicle (OVX); OVX mice treated with alendronate (OVX–ALN); and OVX mice treated with anti-IL-6 receptor (anti-IL-6R) antibody (OVX-IL6i). Sham-operated mice were treated with vehicle. Immediately after surgery, vehicle, ALN, or anti-IL-6R antibody was injected subcutaneously. After a 4-week treatment, mechanical sensitivity was examined using von Frey filaments. The bilateral distal femoral metaphyses and proximal tibial metaphyses were analyzed three-dimensionally with micro-computed tomography. Calcitonin gene-related peptide (CGRP) expression in L3–L5 dorsal root ganglion (DRG) neurons was examined using immunohistochemistry. Ovariectomy induced bone loss and mechanical hyperalgesia in the hindlimbs with upregulation of CGRP expression in the DRG neurons innervating the hindlimbs. ALN treatment prevented bone loss, but anti-IL-6R antibody treatment had no effect on bone morphometry compared with that of the OVX group. However, mechanical hyperalgesia and CGRP expression were significantly decreased in the OVX-IL6i and OVX–ALN groups compared with those in the OVX group. Although anti-IL-6R antibody treatment had no effect on ovariectomy-induced bone loss, the treatment prevented ovariectomy-induced mechanical hyperalgesia in the hindlimbs and suppressed CGRP expression in DRG neurons. The results suggest that IL-6 is one of the causes of postmenopausal osteoporotic pain, and anti-IL-6R antibody might preserve bone health and decrease osteoporotic pain.


Osteoporosis Pain Interleukin-6 



The authors would like to thank Takahiro Iino and Katsura Chiba for excellent technical assistance. No benefits or funds were received in support of the study.

Author Contributions

HW was the principal investigator in this study and was responsible for the original hypothesis, study design, final data analysis, and write-up. SK carried out the set of in vivo studies, performed the in vivo experiments, and reviewed the immunostained sections. GM designed the histochemical protocols and performed immunostaining. NN and YN carried out the in vivo experiments and performed the statistical analysis. AS participated in the design of the study and helped draft the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

Hiroki Wakabayashi, Sho Kato, Nobuto Nagao, Gaku Miyamura, Yohei Naito, and Akihiro Sudo declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

The animal experimentation protocols were approved by the Mie University Animal Care Committee (the approval number: 27-21) and were undertaken in accordance with the ethical guidelines of the National Institute of Health. We followed a randomized, prospective, and controlled animal model design. All efforts were made to minimize animal suffering and the number of animals used.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Orthopaedic SurgeryMie University Graduate School of MedicineTsuJapan

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