Inhibitory effect of quercetin on titanium particle induced endoplasmic reticulum stress related apoptosis and in vivo osteolysis

Abstract

Wear particle induced periprosthetic osteolysis is the main cause of aseptic loosening of orthopedic implants. The aim of the present study is to determine the protective effect of quercetin (QUE) against titanium (Ti) particle induced endoplasmic reticulum stress (ERS) related apoptosis and osteolysis. In the present study, RAW264.7 cells were pretreated with different concentrations (40, 80, and 160 μmol/l) of QUE for 30 min and then treated with Ti particle (5 mg/ml) for 24 h. Cell viability and apoptosis were determined using MTT assay and Annexin V-FITC Apoptosis Detection Kit, respectively. Protein and mRNA expressions of ERS-related genes were examined by Western blot and real-time PCR, respectively. The release of inflammatory cytokines was detected by ELISA. Then, a mouse calvarial osteolysis model was established. Histological sections of calvaria were stained with Hematoxylin-Eosin (H&E) or tartrate-resistant acid phosphatase (TRAP). The results showed that Ti particle reduced cell viability and induced apoptosis in RAW264.7 macrophages. The cytotoxic effects of Ti particle were dramatically inhibited by QUE pretreatment. Interestingly, we found that QUE also significantly reduced Ti particle induced up-regulation of the expression levels of protein kinase RNA-like ER kinase (PERK), inositol-requiring enzyme-1 (IRE1), glucose-regulated protein (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12, and caspase-3 and enhanced the down-regulation of Bcl-2. In addition, QUE decreased Ti particle-induced inflammatory cytokines release from RAW264.7 cells. Moreover, treatment with QUE markedly decreased osteoclast number. In a mouse calvarial osteolysis model, QUE inhibited Ti particle induced osteolysis in vivo by inhibiting osteoclast formation and expressions of ERS-related genes. In conclusion, QUE can protect RAW264.7 cells from Ti particle induced ERS-related apoptosis and suppress calvarial osteolysis in vivo.

Quercetin as an Agent for Protecting the Bone: A Review of the Current Evidence

“…Pretreatment with quercetin restored the downregulated Bcl-2 and Bcl-XL expression as well as the upregulated caspase-3, Bax and cytochrome c expression caused by LPS [75]. Zhang et al (2017) reported that quercetin had anti-apoptotic effects in vitro and in vivo, shown by inhibition of titanium-particle-induced endoplasmic-reticulum-stress-related cell apoptosis. The levels of protein kinase RNA-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1 (IRE1), glucose-regulated protein (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12 and caspase-3 were decreased, whereas Bcl-2 level was increased in the titanium-particle-treated RAW264.7 cells and titanium-particle-implanted calvarium.…”

“…Osteolysis is defined by pathological destruction or disappearance of bone tissues. Zhang et al (2017) determined the protective effects of quercetin against titanium-particle-induced calvarial osteolysis utilising male BALB/c mice. Quercetin at 50 or 100 mg/kg per day inhibited titanium-particle-induced osteolysis by increasing bone area and decreasing osteoclast formation in vivo [45].…”

“…Zhang et al (2017) determined the protective effects of quercetin against titanium-particle-induced calvarial osteolysis utilising male BALB/c mice. Quercetin at 50 or 100 mg/kg per day inhibited titanium-particle-induced osteolysis by increasing bone area and decreasing osteoclast formation in vivo [45]. In another study, female C57BL/6 mice with titanium-particle-induced osteolysis and treated with quercetin (2 or 5 mg/kg/day) for 14 days had higher BV/TV but lower total porosity, erosion area and osteoclast number [46].…”

“…Numerous studies have found that quercetin inhibits the formation of osteoclast-like cells (evidenced by TRAP-positive multinucleated cells), bone resorption pit and F-actin ring formation in RAW264.7 cells, human peripheral-blood mononuclear cells (PBMCs) or bone-marrow macrophages treated with macrophage colony-stimulating factor (M-CSF) and/or RANKL [ 31 , 32 , 45 , 46 , 68 , 74 , 94 , 95 , 96 ]. In another study using highly purified rabbit osteoclasts, treatment with 50 μM quercetin caused a reduction in the total area of resorption pit [ 97 ].…”

Diazoxide Protects against Myocardial Ischemia/Reperfusion Injury by Moderating ERS via Regulation of the miR-10a/IRE1 Pathway

“…GRP78 is a marker protein of ERS and mediates the processing and maturation of nascent proteins. GRP78 is a key regulatory protein that promotes cell protein maturation, cell function, and life under normal growth conditions [ 35 ]. IRE1 is a type I transmembrane protein localized on the ER membrane with both kinase and endonuclease activities.…”