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Resveratrol Increases Osteoblast Differentiation In Vitro Independently of Inflammation

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Abstract

Low-grade inflammation negatively affects bone. Resveratrol is a natural compound proven to possess both anti-inflammatory and bone protective properties. However, it is uncertain if the bone effects are mediated though anti-inflammatory effects. Firstly, we investigated if resveratrol affects proliferation and differentiation of human bone marrow-derived mesenchymal stem cells. Secondly, we investigated if inflammation negatively affects proliferation and differentiation, and if resveratrol counteracts this through anti-inflammatory effects. Mesenchymal stem cells were obtained from bone marrow aspiration in 13 healthy individuals and cultured towards the osteoblast cell lineage. The cells were stimulated with resveratrol, lipopolysaccharide (LPS), LPS + resveratrol, or vehicle (control) for 21 days. Compared to control, resveratrol decreased cell number by 35 % (p < 0.05) and induced differentiation (a 3-fold increase in alkaline phosphatase (p < 0.002), while P1NP and OPG showed similar trends). LPS induced inflammation with a 44-fold increase in interleukin-6 (p < 0.05) and an extremely prominent increase in interleukin-8 production (p < 0.05) relative to control. In addition, LPS increased cell count (p < 0.05) and decreased differentiation (a reduction in P1NP production (p < 0.02)). Co-stimulation with LPS + resveratrol did not reduce interleukin-6 or interleukin-8, but nonetheless, cell count was reduced (p < 0.05) and alkaline phosphatase, P1NP, and OPG increased (p < 0.05 for all). Thus, resveratrol stimulates osteoblast differentiation independently of inflammation.

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Acknowledgments

We thank medical laboratory technicians Pia Hornbek and Lenette Pedersen for their excellent technical assistance.

Contributors

Authors #1, #2, #3, #5, and #6 designed this study. Author #1 prepared the first draft of the paper, and she is guarantor. Author #1 was responsible for statistical analysis of the data. All authors contributed to the experimental work, and revised the paper critically for intellectual content and approved the final version. All authors agree to be accountable for the work and to ensure that any questions relating to the accuracy and integrity of the paper are investigated and properly resolved.

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Authors and Affiliations

  1. Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Tage-Hansens Gade 2, Entrance 3C, 8000, Aarhus, Denmark

    Marie Juul Ornstrup, Torben Harsløf, Lotte Sørensen, Liselotte Stenkjær, Bente Lomholt Langdahl & Steen Bønløkke Pedersen

  2. Department of Clinical Medicine, Aarhus University, Nordre Ringgade 1, 8000, Aarhus, Denmark

    Marie Juul Ornstrup, Torben Harsløf, Bente Lomholt Langdahl & Steen Bønløkke Pedersen

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  1. Marie Juul Ornstrup
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  2. Torben Harsløf
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  4. Liselotte Stenkjær
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  5. Bente Lomholt Langdahl
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Correspondence to Marie Juul Ornstrup.

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Funding

This work was supported by grants from the Elvira and Rasmus Riisfort Foundation, Ejnar Danielsens Foundation, and the AP Møller Maersk Foundation. The study is a part of the research program LIRMOI Research Center (www.LIRMOI.com). LIRMOI is supported by the Danish Council for Strategic Research (Grant 10-093499).

Conflict of Interest

All authors declare that they have no conflict of interest in relation to this work. Author #2 has received speaker honorarium from Amgen, and author #5 is a member of advisory boards and has received speaker honoraria for lectures from Merck, Amgen, Eli Lilly, and UCB. Author #5 also received research grants from Novo Nordisk, Eli Lilly, and Orkla.

Human and Animal Rights and Informed Consent

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in this study.

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Ornstrup, M.J., Harsløf, T., Sørensen, L. et al. Resveratrol Increases Osteoblast Differentiation In Vitro Independently of Inflammation. Calcif Tissue Int 99, 155–163 (2016). https://doi.org/10.1007/s00223-016-0130-x

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  • DOI: https://doi.org/10.1007/s00223-016-0130-x

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