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

  • Ya-lan Pan
  • Yong Ma
  • Yang GuoEmail author
  • Juan Tu
  • Ge-pu Guo
  • Shendu Marshall Ma
  • Su-yang Zheng
  • Peng-cheng Tu
  • Bei-bei Yu
  • Gui-cheng Huang
Original Article



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.


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.


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).


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


Low-intensity pulsed ultrasound Tissue-engineered cartilage Chondrocytes Clematis chinensis Osbeck Transforming growth factor-β/Smad signaling 



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).

Compliance with ethical standards

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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Copyright information

© The Japan Society of Ultrasonics in Medicine 2019

Authors and Affiliations

  • Ya-lan Pan
    • 1
    • 2
  • Yong Ma
    • 1
    • 2
  • Yang Guo
    • 1
    • 2
    Email author
  • Juan Tu
    • 3
  • Ge-pu Guo
    • 3
  • Shendu Marshall Ma
    • 4
  • Su-yang Zheng
    • 1
    • 2
  • Peng-cheng Tu
    • 1
    • 2
  • Bei-bei Yu
    • 5
  • Gui-cheng Huang
    • 1
    • 2
  1. 1.Institute of Traumatology and OrthopedicsNanjing University of Chinese MedicineNanjingChina
  2. 2.Laboratory of New Techniques of Restoration and Reconstruction of Orthopedics and TraumatologyNanjing University of Chinese MedicineNanjingChina
  3. 3.Key Laboratory of Modern Acoustics (MOE), Department of Physics, Collaborative Innovation Center of Advanced MicrostructureNanjing UniversityNanjingChina
  4. 4.Department of Mechanical EngineeringBoston UniversityBostonUSA
  5. 5.Foreign Language CollegeNanjing University of Chinese MedicineNanjingChina

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