In vitro experiment of tetrandrine on the model of osteolysis induced by wear particles around the prosthesis

doi:10.3969/j.issn.2095-4344.3803

Abstract

Abstract: BACKGROUND: There are few domestic studies on the formation and differentiation of osteoclasts induced by wear particles by tetrandrine.
OBJECTIVE: To explore the positive effect of tetrandrine on the osteolysis model of mouse RAW264.7 macrophages induced by titanium particles under different concentrations.
METHODS: Mouse RAW264.7 macrophages were divided into five groups for culture: blank group, experimental group (0.1 g/L titanium particle suspension), and low, medium, and high concentration drug groups (component+0.1, 0.5, and 1.0 μmol/L tetrandrine). CCK-8 assay was used to detect the proliferative activity of RAW264.7. The anti-tartaric acid phosphatase staining method and the phalloidin staining method were used to detect the function and number of mature osteoclasts. The enzyme-linked immunosorbent assay was used to detect the cell supernatant expression of tumor necrosis factor-α and matrix metalloproteinase-9. RT-PCR was applied to detect mRNA expression of activated T cell nuclear factor c1, tumor necrosis factor-α, and matrix metalloproteinase 9. Western blot assay was utilized to detect the levels of phosphorylated proteins of nuclear factor κB p65 and IκBα.
RESULTS AND CONCLUSION: (1) Compared with the blank group, the various concentrations of tetrandrine had no effect on the proliferation of RAW264.7 cells (P > 0.05). (2) In the control group, more bigger, wine red, and F-actin ring stained with fluorescent green multinucleated osteoclasts were observed, and after treatment with different concentrations of tetrandrine, the number of osteoclast cells decreased significantly (P < 0.05); and the size of F-actin ring decreased with the increase of drug concentration. (3) Compared with the blank group, IκBα and nuclear factor κB p65 and their phosphorylation levels, tumor necrosis factor-α, matrix metalloproteinase-9 cell culture supernatant protein expression, activated T cell nuclear factor c1 mRNA, tumor necrosis factor-α mRNA, matrix metalloproteinase-9 mRNA expression levels were significantly increased in the experimental group (P < 0.05). Tetrandrine reduced the expression of the above indexes in a concentration-dependent manner. (4) In summary, tetrandrine can have a positive effect on the in vitro osteolysis model of mouse RAW264.7 macrophages induced by titanium particles by inhibiting the nuclear factor κB signaling pathway.

Key words: tetrandrine, titanium particles, nuclear factor κB, aseptic loosening, osteolysis, artificial prosthesis

CLC Number:

Liu Zige, Liu Xinrui, Li Yan, Song Guorui, Zhang Chen, Chen Desheng. In vitro experiment of tetrandrine on the model of osteolysis induced by wear particles around the prosthesis[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(15): 2358-2363.

Figures/Tables 6

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