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Systemic Inflammation Affects Human Osteocyte-Specific Protein and Cytokine Expression

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Abstract

Bone remodeling can be disturbed in active rheumatoid arthritis (RA), possibly as a result of elevated levels of circulating inflammatory cytokines. Osteocyte-specific proteins and cytokines play a vital role in bone remodeling by orchestrating bone formation and/or bone resorption. Therefore, we aimed to investigate the effect of RA-serum or inflammatory cytokines on expression of human osteocyte-specific proteins and cytokines. Human trabecular bone chips were cultured with RA-serum or inflammatory cytokines for 7-days. Live-dead staining was performed to assess cell viability. Gene expression of osteocyte-specific proteins and cytokines was analyzed by qPCR. Immuno-staining was performed for osteocyte-specific markers. Approximately 60 % of the osteocytes on the bone chips were alive at day-7. Cells in or on the bone chips did express the gene for osteocyte markers SOST, FGF23, DMP1, and MEPE, and the cytokines IL-1β, IL-6, and TNFα at day 0 and 7. Active RA-serum treatment enhanced IL-1β, TNFα, SOST, and DKK1 gene expression. IL-1β treatment enhanced IL-1β, TNFα, IL-6, IL-8, FGF23, and SOST gene expression. TNFα treatment enhanced IL-1β, TNFα, IL-6, IL-8, and FGF23 gene expression. IL-8 treatment enhanced TNFα, IL-8, and FGF23 gene expression. A combination of IL-1β, IL-6, and TNFα treatment synergistically upregulated IL-1β, IL-6, and IL-8 gene expression, as well as enhanced TNFα, OPG, SOST, and FGF23, and inhibited DKK1 gene expression. In conclusion, gene expression of human osteocyte-specific proteins and cytokines was affected by RA-serum, and exogenous recombinant cytokines treatment suggesting that osteocytes could provide a new target to prevent systemic inflammation-induced bone loss in RA.

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Acknowledgments

We thank Mr. Huib W. van Essen and Mrs Jolanda M.A. Hogervorst for technical support. This research was funded by the European Commission through MOVE-AGE, an Erasmus Mundus Joint Doctorate programme (2011-0015).

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    Authors and Affiliations

    1. Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands

      Janak L. Pathak, Astrid D. Bakker & Jenneke Klein-Nulend

    2. Skeletal Biology and Engineering Research Center, KU Leuven, Leuven, Belgium

      Janak L. Pathak, Frank P. Luyten & Patrick Verschueren

    3. Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology (SPST), Tianjin University, Tianjin, China

      Janak L. Pathak

    4. Department of Rheumatology, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands

      Willem F. Lems

    5. Department of Clinical Chemistry, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands

      Nathalie Bravenboer

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    1. Janak L. Pathak
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    Correspondence to Janak L. Pathak.

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    Janak L. Pathak, Astrid D. Bakker, Frank P. Luyten, Patrick Verschueren, Willem F. Lems, Jenneke Klein-Nulend, and Nathalie Bravenboer declare that they have no conflict of interest.

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    The protocol was approved by the Ethical Review Board of the VU University Medical Center, Amsterdam, The Netherlands, and all subjects gave informed consent.

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    J. Klein-Nulend and N. Bravenboer have equally contributed to this study and also shared last authorship.

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    Pathak, J.L., Bakker, A.D., Luyten, F.P. et al. Systemic Inflammation Affects Human Osteocyte-Specific Protein and Cytokine Expression. Calcif Tissue Int 98, 596–608 (2016). https://doi.org/10.1007/s00223-016-0116-8

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    • DOI: https://doi.org/10.1007/s00223-016-0116-8

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