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Effects of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on transforming growth factor-β/Smad signaling in rabbit articular chondrocytes

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Abstract

Purpose

Clematis chinensis Osbeck (CCO) is an essential herb that has been shown to promote the biological functions of cartilage cells. In this study, we aimed to explore whether and how low-intensity pulsed ultrasound (LIPUS) enhanced CCO delivery into chondrocytes and stimulated biological activity in vitro.

Methods

Chondrocytes were isolated from knee articular cartilage of 2-week-old rabbits and treated with LIPUS plus CCO or recombinant transforming growth factor beta 1 (TGF-β1; 0.5 ng/mL), with or without anti-TGF-β1 antibodies (10 μg/mL), for 3 days. Cell proliferation was assessed by Cell-Counting Kit-8 assays. Immunocytochemistry, western blotting, and quantitative polymerase chain reaction were applied to detect the expression of type II collagen and some molecules in the TGF-β1 signal pathway.

Results

LIPUS plus 0.1 mg/mL CCO solution promoted chondrocyte proliferation and type II collagen and TGF-β1 expression synergistically in vitro (P < 0.05). In addition, treatment with anti-TGF-β1 antibodies blocked this effect (P < 0.01), but not completely. CCO plus LIPUS also showed more enhanced effects on promoting TGF-β receptor II and Smad2 signaling and reducing Smad7 signaling than either intervention separately (P < 0.05).

Conclusions

CCO plus LIPUS promoted extracellular matrix deposition by accelerating the TGF-β/Smad-signaling pathway in chondrocytes.

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Acknowledgements

This study was financially supported by the National Natural Science Foundation of China (No. 81673995), Natural Science Foundation for Youths of Jiangsu Province, China (No. BK20151007), Natural Science Foundation of Jiangsu Province, China (No. BK2011812, No. BK20161047), and Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX17_1308).

Author information

Author notes

  1. Ya-lan Pan and Yong Ma contributed equally to this study.

Authors and Affiliations

  1. Institute of Traumatology and Orthopedics, Nanjing University of Chinese Medicine, 138 Xianlin Road, Nanjing, 210023, China

    Ya-lan Pan, Yong Ma, Yang Guo, Su-yang Zheng, Peng-cheng Tu & Gui-cheng Huang

  2. Laboratory of New Techniques of Restoration and Reconstruction of Orthopedics and Traumatology, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Ya-lan Pan, Yong Ma, Yang Guo, Su-yang Zheng, Peng-cheng Tu & Gui-cheng Huang

  3. Key Laboratory of Modern Acoustics (MOE), Department of Physics, Collaborative Innovation Center of Advanced Microstructure, Nanjing University, Nanjing, 210093, China

    Juan Tu & Ge-pu Guo

  4. Department of Mechanical Engineering, Boston University, Boston, MA, 02215, USA

    Shendu Marshall Ma

  5. Foreign Language College, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Bei-bei Yu

Authors
  1. Ya-lan Pan
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  2. Yong Ma
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  3. Yang Guo
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  4. Juan Tu
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Corresponding author

Correspondence to Yang Guo.

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Ethical statements

All experimental procedures were approved by the institutional and local committee on the care and use of animals of Nanjing University of Chinese Medicine.

Conflict of Interest

The authors declare that they have no conflicts of interest.

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Pan, Yl., Ma, Y., Guo, Y. et al. Effects of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on transforming growth factor-β/Smad signaling in rabbit articular chondrocytes. J Med Ultrasonics 46, 177–186 (2019). https://doi.org/10.1007/s10396-018-0920-z

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  • DOI: https://doi.org/10.1007/s10396-018-0920-z

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