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Vasoactive Intestinal Peptide Promotes Fracture Healing in Sympathectomized Mice

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Abstract

Vasoactive intestinal peptide (VIP) as a neuromodulator and neurotransmitter played a significant role in modulating bone homeostasis. Our previous study reported an essential role of VIP in in vitro BMSCs osteogenesis and in vivo bone defect repair. VIP was also revealed to have a promoting effect on embryonic skeletal element development. However, the role of VIP in fracture healing is not known yet. We hypothesized that the disorder of sympathetic nervous system impairs bone structure and fracture healing, whereas VIP may rescue the sympathetic inhibition effects and promote fracture healing. We employed a 6-hydroxydopamine (6-OHDA) induced sympathectomy mice model (sympathectomized mice), in which successful sympathetic inhibition was confirmed by a decreased level of norephedrine (NE) in the spleen. In the sympathectomized mice, the femoral micro-architecture, bone density and mechanical properties were all impaired compared to the vehicle control mice. The femoral fracture was created in the vehicle or sympathectomized mice. Vehicle mice were locally injected with PBS as a negative control, and the sympathectomized mice were treated with injection of PBS or VIP. VIP expression at the fracture site was significantly decreased in sympathectomized mice. The fracture healing was repressed upon 6-OHDA treatment and rescued by VIP treatment. Micro-CT examination showed that the femoral bone micro-architecture at the fracture sites and mechanical properties were all impaired. Simultaneously, the expression level of osteogenic markers OCN and OPN were reduced in sympathectomized mice compared with vehicle group. While the VIP treatment rescued the repression effects of 6-OHDA on bone remodeling and significantly promoted bone quality and mechanical properties as well as increased osteogenesis marker expression in the sympathectomized mice. VIP administration promoted bone fracture healing by inhibiting bone resorption, making it a putative new alternative treatment strategy for fracture healing.

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Acknowledgements

This work was partially supported by grants from the National Natural Science Foundation of China (Grant No. 81772322); Hong Kong Government Research Grant Council, General Research Fund (Grant Nos. 14120118, 14160917, C7030-18G and T13-402/17-N); Hong Kong Innovation Technology Commission Funds (Grant No. PRP/050/19FX); Hong Kong Medical Research Funds (Grant Nos. 16170951 and 17180831). This study also received support from the SMART program, Lui Che Woo Institute of Innovative Medicine, The Chinese University of Hong Kong.

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Authors and Affiliations

  1. Department of Orthopaedics & Traumatology, Faculty of Medicine, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong SAR, NT, People’s Republic of China

    Liu Shi, Yang Liu, Zhengmeng Yang, Tianyi Wu, Hiu Tung Lo, Jia Xu, Jiajun Zhang, Weiping Lin, Jinfang Zhang & Lu Feng

  2. Department of Orthopaedics, School of Medicine, Zhongda Hospital, Southeast University, No. 87 Ding Jia Qiao, Nanjing, 210009, Jiangsu, People’s Republic of China

    Liu Shi

  3. Trauma Center, School of Medicine, Zhongda Hospital, Southeast University, No. 87 Ding Jia Qiao, Nanjing, 210009, Jiangsu, People’s Republic of China

    Liu Shi

  4. Department of Orthopaedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai, People’s Republic of China

    Tianyi Wu

  5. Stem Cells and Regeneration Laboratory, Faculty of Medicine, Prince of Wales Hospital, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Shatin, People’s Republic of China

    Jia Xu

  6. Key Laboratory of Orthopaedics and Traumatology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, The First Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China

    Jinfang Zhang

  7. Laboratory of Orthopaedics & Traumatology, Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China

    Jinfang Zhang

  8. MOE Key Laboratory for Regenerative Medicine, School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, People’s Republic of China

    Gang Li

  9. Department of Orthopaedics & Traumatology, Faculty of Medicine, Prince of Wales Hospital, The Chinese University of Hong Kong, Room 501, Li Ka Shing Medical Sciences Building, Shatin, Hong Kong SAR, NT, People’s Republic of China

    Gang Li

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  1. Liu Shi
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  2. Yang Liu
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  3. Zhengmeng Yang
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  10. Lu Feng
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  11. Gang Li
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Contributions

GL and LF led and supervised all the experiments. GL, JFZ, and LF designed experiments. LS, YL, ZMY, TYW, JX, JJZ and WPL conducted experiments. LS and LF analyzed data. LS, HTL, LF, and GL prepared, wrote and revised the manuscript. All authors reviewed and approved the manuscript.

Corresponding authors

Correspondence to Lu Feng or Gang Li.

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Human and Animal Rights and Informed Consent

No human specimen were applied in this study. The animal experiments were performed in accordance with the Code of Ethics of the World Medical Association. The ethics approval (14/118/MIS) of this study was obtained from the Animal Research Ethics Committee (AEEC) of The Chinese University of Hong Kong.

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Shi, L., Liu, Y., Yang, Z. et al. Vasoactive Intestinal Peptide Promotes Fracture Healing in Sympathectomized Mice. Calcif Tissue Int 109, 55–65 (2021). https://doi.org/10.1007/s00223-021-00820-9

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