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Mechanical stress-induced apoptosis of nucleus pulposus cells: an in vitro and in vivo rat model

  • Original Article
  • Published:
Journal of Orthopaedic Science

Abstract

Background

Un-physiological loads play an important role in the degenerative process of inter-vertebral discs (IVD). In this study, we used an in vitro and in vivo rat model to investigate the mechanism of nucleus pulposus (NP) cells apoptosis induced by mechanical stress.

Methods

Static compressive load to IVDs of rat tails was used as the in vivo model. For the in vitro model, NP cells were tested under the physiological and un-physiological loading. For histological examination, apoptotic index study, and apoptotic gene expression, we also selected cytokines [bone morphogenetic protein (BMP)-2/7, insulin-like growth factor (IGF)-1, platelet-derived growth factor (PDGF)] to be analyzed.

Results

Under mechanical loading, cellular density was significantly decreased, but there was an increase of TUNEL positive cells and apoptosis index. In a dose-dependent manner; the necrosis became apparent in the un-physiologic strain. The selected cytokines (BMP-2/7, IGF-1, PDGF) can significantly reduce the percentage of apoptotic and necrotic cells.

Conclusions

We conclude that the intrinsic (mitochondrial) apoptotic pathway plays an important role in the compressive load-induced apoptosis of NP cells. Combination therapy reducing the mechanical load and selected cytokines (BMP-2/7, IGF-1 and PDGF) may have considerable promise in the treatment of spine disc degeneration.

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Acknowledgments

The authors wish to acknowledge The National Science Council, Taiwan, ROC and Research Fund of Hsin-Chu General Hospital, Department of Health, Executive Yuan, Taiwan, ROC for their funding assistance for this work. We also thank Ms. Margaret, ManGer Sun for her help and valuable suggestions.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Yi-Jie Kuo

  2. Graduate Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan, ROC

    Lien-Chen Wu

  3. Department of Orthopedic Surgery, School of Medicine, College of Medicine, National Taiwan University, 1, Jen-Ai Road, Section 1, Taipei, 10051, Taiwan, ROC

    Jui-Sheng Sun

  4. Department of Orthopedic Surgery, National Taiwan University Hospital Hsin-Chu Branch, No.25, Lane 442, Sec. 1, Jingguo Rd., Hsinchu City, 30059, Taiwan, ROC

    Jui-Sheng Sun

  5. Department of Surgery, National Taiwan University Hospital Hsin-Chu Branch, Hsinchu City, Taiwan, ROC

    Ming-Hong Chen

  6. Department of Biology, Western University, London, ON, N6A 5C1, Canada

    Man-Ger Sun

  7. Department of Orthopedic Surgery, Taipei Medical University-Shuang-Ho Hospital, No.291, Jhongjheng Rd., Jhonghe City, New Taipei, 23561, Taiwan, ROC

    Yang-Hwei Tsuang

Authors
  1. Yi-Jie Kuo
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  2. Lien-Chen Wu
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  3. Jui-Sheng Sun
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  4. Ming-Hong Chen
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  5. Man-Ger Sun
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Corresponding authors

Correspondence to Jui-Sheng Sun or Yang-Hwei Tsuang.

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776_2013_510_MOESM1_ESM.tif

Fig. S Histological and Immuno-histochemical TUNEL stain evaluation of stab injury and static compression-loading on the inter-vertebral disc. Upper row: In the normal inter-vertebral disc of the control sample, the gelatinous nucleus pulposus contains a cellular interior with proteoglycan around the perimeter, sometimes with a capsule around them. At the time point of 1 day after stab injury, there were degenerative changes at the disc tissue. At the time point of 3 days after stab injury, the stab-injured discs showed significant sparse cellular density, clusters of nucleus pulposus cells and abundant extracellular matrix expression. Normal morphology was observed in the majority of discs loaded with 6.4N-1day which was similar to the control group. In the 6.4N-3 day group, there were structural changes in the nucleus, consisting of increased extracellular matrix and a decreased area of the cellular region. Similar changes were observed in the 20.8N-1day’s disc. The changes were more pronounced as significant decreased cellular density, increased extracellular matrix expression and clusters of nucleus pulposus cells were noted in the 20.8N-3days’ discs. Lower Row: In this study, no matter the compressive load or what were the stabbing-injured discs, they showed a significant increase of TUNEL positive cells, indicating evident apoptosis in comparison to the control group in a dose dependent pattern (TIFF 923 kb)

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Kuo, YJ., Wu, LC., Sun, JS. et al. Mechanical stress-induced apoptosis of nucleus pulposus cells: an in vitro and in vivo rat model. J Orthop Sci 19, 313–322 (2014). https://doi.org/10.1007/s00776-013-0510-2

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  • DOI: https://doi.org/10.1007/s00776-013-0510-2

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