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Functional block of IL-17 cytokine promotes bone healing by augmenting FOXO1 and ATF4 activity in cortical bone defect model

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Abstract

Summary

We determine the effect of interleukin (IL)-17 neutralizing antibody on new bone regeneration. Anti-IL-17 antibody promoted new bone regeneration in cortical bone defect model by augmenting FOXO1 and ATF4 activity thereby decreasing oxidative stress. Our study demonstrates the bone healing and regeneration potential of neutralizing IL-17antibody in osteoporotic fractures.

Introduction

The immune system plays important role in the fracture healing process. However, fracture healing is prolonged in disorders associated with systemic inflammation. Fracture healing is decelerated in osteoporosis, condition linked with systemic inflammation. Bone regeneration therapies like recombinant human BMP2 are associated with serious side effects. Studies have been carried out where agents like denosumab and infliximab enhance bone regeneration in osteoporotic conditions. Our previous studies show the osteoprotective and immunoprotective effects of neutralizing IL-17 antibody. Here, we determine the effect of IL-17 neutralizing antibody on new bone regeneration and compare its efficacy with known osteoporotic therapies.

Methods

For the study, female BALB/c mice were ovariectomized or sham operated and left for a month followed by a 0.6-mm drill-hole injury in femur mid-diaphysis. The treatment was commenced next day onwards with anti-IL-17, anti-RANKL (Receptor activator of nuclear factor kappa-B ligand), parathyroid hormone (PTH), or alendronate for a period of 3, 10, or 21 days. Animals were then autopsied, and femur bones were dissected out for micro-CT scanning, confocal microscopy, and gene and protein expression studies.

Results

Micro-CT analysis showed that anti-IL-17 antibody promoted bone healing at days 10 and 21, and the healing effect observed was significantly better than Ovx, anti-RANKL antibody, and ALN, and equal to PTH. Anti-IL-17 also enhanced new bone regeneration as assessed by calcein-labeling studies. Additionally, anti-IL-17 therapy enhanced expression of osteogenic markers and decreased oxidative stress at the injury site.

Conclusion

Overall, our study demonstrates bone healing and regeneration potential of neutralizing IL-17 antibody in osteoporotic fractures.

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Acknowledgements

We would like to acknowledge Centre for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI), Council of Scientific and Industrial Research (CSIR), Government of India for generous funding. MD would like to acknowledge the Department of Biotechnology (DBT), Government of India; KBS would like to acknowledge the Indian Council of Medical Research (ICMR), Government of India; and RP would like to acknowledge the University Grants Commission (UGC) for grant of fellowships. Mr. Geet K Nagar is acknowledged for helping in histology work. Dr. Kavita Singh is acknowledged for helping in confocal microscopy facility of SAIF Division.

Supporting grants

Centre for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI), Council of Scientific and Industrial Research (CSIR), Government of India. CDRI communication no: 9464.

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Authors and Affiliations

  1. Division of Endocrinology and Centre for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI), CSIR-Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Lucknow, India

    M. Dixit, K. B. Singh, R. Prakash & D. Singh

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  1. M. Dixit
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  2. K. B. Singh
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  3. R. Prakash
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  4. D. Singh
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Correspondence to D. Singh.

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Author’s roles

DS and MD conceived and designed the experiments. MD carried out all the experiments. KR and RP helped with the animal-related experiments. MD and DS analyzed the data. DS wrote the manuscript.

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Dixit, M., Singh, K.B., Prakash, R. et al. Functional block of IL-17 cytokine promotes bone healing by augmenting FOXO1 and ATF4 activity in cortical bone defect model. Osteoporos Int 28, 2207–2220 (2017). https://doi.org/10.1007/s00198-017-4012-5

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