Mechanism underlying the effect of Liuwei Dihuang Pill on osteolysis and osteogenesis induced by titanium particles

doi:10.12307/2023.880

Abstract

Abstract: BACKGROUND: At present, a large number of studies have found that Liuwei Dihuang Pill can be used to treat osteoporosis, but there are few related studies on the differentiation and mineralization of osteoblasts induced by wear particles using Liuwei Dihuang Pill.
OBJECTIVE: To investigate the positive effect of different concentrations of Liuwei Dihuang Pill-containing serum on titanium particle-induced mouse MC3T3-E1 osteoblast in vitro osteolysis model.
METHODS: Drug-containing serum was extracted after oral administration of Liuwei Dihuang Pill. The best concentration of Liuwei Dihuang Pill-containing serum and titanium particles on the viability of MC3T3-E1 cells was screened. MC3T3-E1 cells were divided into three groups. The blank group was given osteoblastic differentiation culture. The model group was given titanium particles (5 μg/mL) ossification culture. The drug-containing serum group was given titanium particles (5 μg/mL) + Liuwei Dihuang Pill-containing serum (10%, 15% and 20% doses). Osteoblast viability was detected by CCK-8 assay. Cell alkaline phosphatase activity was detected by alkaline phosphatase staining. Cell mineralization was detected by silver nitrate (Von Kossa) and alizarin red staining. Expression levels of bone differentiation-related genes Runx-2, Osterix, Ocn, Axin, Alp, and Opn were detected by qRT-PCR. Wnt/β-catenin signaling pathways β-catenin, p-GSK-3β, GSK-3β, Runx2 and Osterix protein expression levels were detected by western blot assay.
RESULTS AND CONCLUSION: (1) Liuwei Dihuang Pill-containing serum culture reversed the decrease in alkaline phosphatase activity of MC3T3E-1 cells induced by titanium particles, increased the alizarin red staining and calcification of MC3T3E-1 cells, increased the expression of osteogenesis-related genes in MC3T3E-1 cells, and increased the expression of proteins related to the Wnt/β-catenin signaling pathway. (2) These findings indicate that Liuwei Dihuang Pill-containing serum can reverse the inhibitory effect of titanium particles on the differentiation and mineralization of osteoblasts, upregulate the expression of osteogenesis-related genes, and its mechanism is related to the regulation of Wnt/β-catenin signaling pathway, suggesting that Liuwei Dihuang Pill is expected to become an effective drug for preventing aseptic loosening of artificial joints.

Key words: Liuwei Dihuang Pill-containing serum, osteoblast, osteolysis, Wnt/β-catenin, titanium particle

CLC Number:

Zhu Zhiqi, Yuan Sijie, Zhang Zilin, Ji Shijie, Meng Mingsong, Yan Anming, Han Jing. Mechanism underlying the effect of Liuwei Dihuang Pill on osteolysis and osteogenesis induced by titanium particles[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(3): 392-397.

Figures/Tables 6

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