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Long-Term High-Dose Resveratrol Supplementation Reduces Bone Mass and Fracture Strength in Rats

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Abstract

Resveratrol (RSV) is a natural polyphenolic compound. A recent study suggests a positive effect on BMD in men; however, the underlying changes in microstructure and strength remain unknown. We aimed to investigate the effects of RSV on the skeleton in hindlimb-immobilized and non-immobilized rats. Seventy-two female Wistar rats were divided into six groups. Two baseline (BSL) groups underwent short-term diet intervention for 4 weeks before sacrifice [phytoestrogen-deficient diet (PD) (BSL + PD) or RSV diet (600 mg/kg body weight/day) (BSL + RSV)]. Four groups were injected in the right hindlimb with botulinum toxin (BTX) (immobilized) or saline (non-immobilized), and fed either PD diet or RSV diet 4 weeks pre-injection and 6 weeks post-injection before sacrifice (BTX + PD, BTX + RSV, PD, and RSV, respectively). DXA, µCT, dynamic histomorphometry, and mechanical tests were performed. Short-term RSV treatment did not affect bone parameters, whereas long-term RSV exposure had a consistent negative impact on non-immobilized rats (RSV vs. PD); whole femoral aBMD (p = 0.01) and distal femoral metaphyseal Tb.N (p = 0.01), Tb.Sp (p = 0.02), and BV/TV (p = 0.07). At the femoral mid-diaphysis, RSV increased periosteal resorption (p = 0.01) and increased endosteal formation (p = 0.02), while mineralization was unaffected. In addition, RSV reduced femoral mid-diaphyseal three-point bending strength (p = 0.03) and stiffness (p = 0.04). BTX-induced immobilization resulted in significant bone loss and reduced bone strength; however, RSV supplementation was unable to prevent this. In conclusion, long-term high-dose RSV reduced bone mass and fracture strength and did not prevent immobilization-induced bone loss in rats.

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Acknowledgements

The authors are grateful for the excellent technical assistance of Jytte Utoft and collaboration with Kim Henriksen (Nordic Bioscience A/S, Herlev, Denmark). We thank Visiopharm for the contribution to the newCAST stereology software system. The μCT scanner was kindly donated by the VELUX Foundation.

Funding

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).

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Authors and Affiliations

  1. Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus C, Denmark

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

  2. Department of Clinical Medicine, Aarhus University, Aarhus, Denmark

    Marie Juul Ornstrup, Bente Lomholt Langdahl & Steen Bønløkke Pedersen

  3. Department of Biomedicine, Aarhus University, Aarhus, Denmark

    Annemarie Brüel & Jesper Skovhus Thomsen

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  1. Marie Juul Ornstrup
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  4. Torben Harsløf
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  5. Bente Lomholt Langdahl
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  6. Steen Bønløkke Pedersen
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Contributions

All authors designed this study. MJO prepared the first draft of the paper, and she is guarantor. MJO was responsible for statistical analysis of the data. MJO, AB, and JST contributed to the experimental work, and all authors 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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Correspondence to Marie Juul Ornstrup.

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Conflict of interest

Marie Juul Ornstrup, Annemarie Brüel, Jesper Skovhus Thomsen, Torben Harsløf, Bente Lomholt Langdahl, Steen Bønløkke Pedersen declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

All applicable international, national, and institutional guidelines for the care and use of animals were followed, and all experiments and procedures complied with the guiding principles in the Guide for the Care and Use of Laboratory Animals [45] and was approved by the Danish Animal Experiments Inspectorate (2012-15-2934-00769).

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Ornstrup, M.J., Brüel, A., Thomsen, J.S. et al. Long-Term High-Dose Resveratrol Supplementation Reduces Bone Mass and Fracture Strength in Rats. Calcif Tissue Int 102, 337–347 (2018). https://doi.org/10.1007/s00223-017-0344-6

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