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In vivo and in vitro evaluation of effects of Mg-6Zn alloy on apoptosis of common bile duct epithelial cell

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Abstract

Biodegradable magnesium alloy implants have attracted much attention because of their excellent biocompatibility and good mechanical properties. However, effects of Mg alloy on cell apoptosis remain unclear. The aim of this study was to investigate the effects of the Mg-6Zn alloy on the apoptosis and necrosis of common bile duct (CBD) epithelial cells. In the in vitro experiments, primary mouse extrahepatic bile epithelial cells (MEBECs) were exposed to Mg-6Zn alloy extracts with different concentrations (0, 40, 80, and 100 %). Flow cytometry analysis indicated that low concentration Mg-6Zn extract can induce apoptosis of MEBECs, and high concentration Mg-6Zn extracts may relate to necrosis and/or ‘apoptotic necrosis’. Real-time PCR results showed that when MEBECs were treated with 40 % extracts for 3 days, the relative apoptotic genes including Bax, Bax/Bcl-2 ratio, NF-κB and caspase-3 were higher than those in the control group. In the in vivo experiments, Mg-6Zn alloy stents were implanted into rabbits’ CBD for 1, 2, 3 weeks, respectively. Based on the hematoxylin and eosin (H&E) staining of peri-implant CBD tissue, no apoptotic bodies and necrotic cells were observed. Results of immunohistochemical staining also showed Mg-6Zn stents did not increase expression levels of apoptosis related gene such as Bax, Bcl-2, Bax/Bcl-2 ratio, TNF-α, NF-κB and caspase-3 in CBD, which indicating Mg-6Zn did not induce significant apoptosis in the in vivo experiments. The different results of in vitro and in vivo experiment may result from the low corrosion rate of Mg-6Zn alloy stents in vivo and local Mg2+ ion concentration in CBD.

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Acknowledgments

This work was supported by the National Basic Research Program of China (973 Program, No. 2012CB619102), the National Natural Science Foundation of China (No. 30901422 and No. 51271117) and Shanghai Jiao Tong University Interdisciplinary (Biomedical Engineering) Research Fund (No. YG2010MS45). Shanghai Jiao Tong University School of Medicine Science and Technology Fund (No. 09XJ21005).

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Authors and Affiliations

  1. Department of General Surgery, Nanjing Medical University affiliated Wuxi No. 2 People’s Hospital, Nanjing, Jiangsu, 214002, People’s Republic of China

    Yigang Chen

  2. Department of General Surgery, Sixth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 600 Yishan Road, Shanghai, 200233, People’s Republic of China

    Jun Yan, Xiaohu Wang, Song Yu, Zigang Wang & Qi Zheng

  3. State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, No. 800 DongChuan Road, Shanghai, 200240, People’s Republic of China

    Xiaonong Zhang & Changli Zhao

  4. Suzhou Origin Medical Technology Co. Ltd, 2 Haicheng Road, Changshu Economic and Technology Development Zone, Suzhou, Jiangsu, 215513, People’s Republic of China

    Shaoxiang Zhang

  5. Department of Advanced Materials and Nanotechnology, College of Engineering, Peking University, Beijing, 100871, People’s Republic of China

    Yufeng Zheng

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  1. Yigang Chen
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  2. Jun Yan
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  3. Xiaohu Wang
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  4. Song Yu
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  6. Xiaonong Zhang
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  7. Shaoxiang Zhang
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  8. Yufeng Zheng
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  9. Changli Zhao
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  10. Qi Zheng
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Correspondence to Changli Zhao or Qi Zheng.

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Jun Yan contributed equally to this work.

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Chen, Y., Yan, J., Wang, X. et al. In vivo and in vitro evaluation of effects of Mg-6Zn alloy on apoptosis of common bile duct epithelial cell. Biometals 27, 1217–1230 (2014). https://doi.org/10.1007/s10534-014-9784-x

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  • DOI: https://doi.org/10.1007/s10534-014-9784-x

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