Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (38): 6110-6115.

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Effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation and expression of type II collagen and transforming growth factor-beta1 of rabbit knee articular chondrocytes

Ma Yong1, 2, Guo Yang1, Tu Juan3, Cheng Ji-hua1, Guo Ge-pu3, Huang Gui-cheng1   

  1. 1Institute of Traumatology & Orthopedics, Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu Province, China; 2Department of Traumatology & Orthopedics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, Jiangsu Province, China; 3Institute of Acoustics, Nanjing University, Nanjing 210093, Jiangsu Province, China
  • Online:2014-09-10 Published:2014-09-10
  • Supported by:

    Natural Science Foundation of Jiangsu Province, No. BK2011812; the “Six Top Talents” of Jiangsu Province, No. 2010-WS-082

Abstract:

BACKGROUND: Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II collagen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cell membrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals.
OBJECTIVE: To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II collagen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage. 
METHODS: The rabbit knee articular chondrocytes were isolated and cultured in vitro. Cells of the second generation in logarithmic growth phase were randomly divided into four groups: control group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO + LIPUS group. After cells were cultured under different conditions for 3 days, cell proliferation was measured with Cell Count Kit-8 assay. The secretion of type II collagen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis.
RESULTS AND CONCLUSION: The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P < 0.05), while the CCO + LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II collagen in cartilage cells were significantly increased in CCO + LIPUS group, compared with control group (P < 0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO + LIPUS group, and the relative gray value was significantly higher than the other groups (P < 0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II collagen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.



中国组织工程研究
杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程


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Key words: chondrocytes, Chinese medicine, ultrasound treatment, collagen, type II, tumor necrosis factor-beta1

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