NOX4 blockade suppresses titanium nanoparticle-induced bone destruction via activation of the Nrf2 signaling pathway

Periprosthetic osteolysis (PPO) induced by wear particles is the most severe complication of total joint replacement; however, the mechanism behind PPO remains elusive. Previous studies have shown that osteocytes play important roles in wear-particle-induced osteolysis. In this study, we investigated the effects of connexin 43 (Cx43) on the regulation of osteocyte-to-osteoblast differentiation. We established an in vivo murine model of calvarial osteolysis induced by titanium (Ti) particles. The osteolysis characteristic and osteogenesis markers in the osteocyte-selective Cx43 (CKO)-deficient and wild-type (WT) mice were observed. The calvarial osteolysis induced by Ti particles was partially attenuated in CKO mice. The expression of β-catenin and osteogenesis markers increased significantly in CKO mice. In vitro, the osteocytic cell line MLO-Y4 was treated with Ti particles. The co-culturing of MLO-Y4 cells with MC3T3-E1 osteoblastic cells was used to observe the effects of Ti-treated osteocytes on osteoblast differentiation. When Cx43 of MLO-Y4 cells was silenced or overexpressed, β-catenin was detected. Additionally, co-immunoprecipitation detection of Cx43 and β-catenin binding in MLO-Y4 cells and MC3T3-E1 cells was performed. Finally, β-catenin expression in MC3T3-E1 cells and osteoblast differentiation were evaluated after 18α-glycyrrhetinic acid (18α-GA) was used to block the intercellular communication of Cx43 between MLO-Y4 and MC3T3-E1 cells. Ti particles increased Cx43 expression and decreased β-catenin expression in MLO-Y4 cells. The silencing of Cx43 increased the β-catenin expression, and the over-expression of Cx43 decreased the β-catenin expression. In the co-culture model, Ti treatment of MLO-Y4 cells inhibited the osteoblastic differentiation of MC3T3-E1 cells and Cx43 silencing in MLO-Y4 cells attenuated the inhibitory effects on osteoblastic differentiation. With Cx43 silencing in the MLO-Y4 cells, the MC3T3-E1 cells, co-cultured alongside MLO-Y4, displayed decreased Cx43 expression, increased β-catenin expression, activation of Runx2, and promotion of osteoblastic differentiation in vitro co-culture. Finally, Cx43 expression was found to be negatively correlated to the activity of the Wnt signaling pathway, mostly through the Cx43 binding of β-catenin from its translocation to the nucleus. The results of our study suggest that Ti particles increased Cx43 expression in osteocytes and that osteocytes may participate in the regulation of osteoblast function via the Cx43 during PPO.

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“…Wear particles induce ROS-mediated immune responses ( 24 ) and are crucial for RANKL-induced osteoclast differentiation ( 38 ). Elevated ROS levels decrease Nrf2 expression, thereby contributing to increased Nfatc1 expression during osteoclast formation ( 39 , 40 ). Accordingly, ROS signaling is indicated to be a suitable target for the pharmacological treatment of bone loss ( 30 ).…”

Section: Discussionmentioning

confidence: 99%

Britanin inhibits titanium wear particle‑induced osteolysis and osteoclastogenesis

Kim,

Lim,

Ihn

et al. 2023

Mol Med Rep

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Wear particle-induced osteolysis is a serious complication that occurs in individuals with titanium (Ti)-based implants following long-term usage due to loosening of the implants. The control of excessive osteoclast differentiation and inflammation is essential for protecting against wear particle-induced osteolysis. The present study evaluated the effect of britanin, a pseudoguaianolide sesquiterpene isolated from Inula japonica , on osteoclastogenesis in vitro and Ti particle-induced osteolysis in vivo . The effect of britanin was examined in the osteoclastogenesis of mouse bone marrow-derived macrophages (BMMs) using TRAP staining, RT-PCR, western blotting and immunocytochemistry. The protective effect of britanin was examined in a mouse calvarial osteolysis model and evaluated using micro-CT and histomorphometry. Britanin inhibited osteoclast differentiation and F-actin ring formation in the presence of macrophage colony-stimulating factor and receptor activator of nuclear factor kB ligand in BMMs. The expression of osteoclast-specific marker genes, including tartrate-resistant acid phosphatase, cathepsin K, dendritic cell-specific transmembrane protein, matrix metallopeptidase 9 and nuclear factor of activated T-cells cytoplasmic 1, in the BMMs was significantly reduced by britanin. In addition, britanin reduced the expression of B lymphocyte-induced maturation protein-1, which is a transcriptional repressor of negative osteoclastogenesis regulators, including interferon regulatory factor-8 and B-cell lymphoma 6. Conversely, britanin increased the expression levels of anti-oxidative stress genes, namely nuclear factor erythroid-2-related factor 2, NAD(P)H quinone oxidoreductase 1 and heme oxygenase 1 in the BMMs. Furthermore, the administration of britanin significantly reduced osteolysis in a Ti particle-induced calvarial osteolysis mouse model. Based on these findings, it is suggested that britanin may be a potential therapeutic agent for wear particle-induced osteolysis and osteoclast-associated disease.

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NOX4 blockade suppresses titanium nanoparticle-induced bone destruction via activation of the Nrf2 signaling pathway

Cited by 7 publications

References 61 publications

Osteocytes Exposed to Titanium Particles Inhibit Osteoblastic Cell Differentiation via Connexin 43/β-Catenin Pathway

Osteocytes Exposed to Titanium Particles Inhibit Osteoblastic Cell Differentiation via Connexin 43/β-Catenin Pathway

Osteocytes Exposed to Titanium Particles Inhibit Osteoblastic Cell Differentiation via Connexin 43

Britanin inhibits titanium wear particle‑induced osteolysis and osteoclastogenesis

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