Department of Orthopaedics, Fuzhou Second Hospital affiliated to Xiamen University, Fujian Province 350007, China.
Biomed Res Int. 2021 Mar 4;2021:5578088. doi: 10.1155/2021/5578088. eCollection 2021.
This study is aimed at studying the effect of zoledronate (ZOL) on the differentiation of osteoclast precursor RAW264.7 cells induced by titanium (Ti) particles and explores the possibility of preventing and treating periprosthetic osteoporosis using ZOL.
RAW264.7 cells were cultured in vitro. Ti particles were prepared. The cell proliferation curve of RAW264.7 cells was plotted using the MTT assay to find the best concentration of ZOL for intervention. The cells were divided into three groups: control, Ti particles, and Ti particles+ZOL. The cell morphology was observed using tartaric acid-resistant acid phosphatase (TRAP) staining, and the activity of TRAP in cell supernatant was determined using the biochemical method. The number of bone resorption lacunae was detected using toluidine blue staining. The mRNA expression of , , , and was detected using real-time polymerase chain reaction. The protein expression of RANK, NFATcl, and MMP-9 was detected using Western blot analysis.
Ti particles stimulated the differentiation of RAW264.7 cells into osteoclasts. They also increased the activity of TRAP, number of bone resorption lacunae, and mRNA and protein expression of RANK, NFATcl, and MMP-9. However, ZOL could suppress the effect of TI particles on the osteoclast differentiation of RAW264.7 cells.
ZOL could effectively inhibit the differentiation of RAW264.7 cells into osteoclasts induced by Ti particles, decrease the activity of TRAP, reduce the number of bone resorption lacunae, and decrease the mRNA and protein expression of RANK, NFATcl, and MMP-9. Hence, it may be a promising candidate for preventing and treating periprosthetic osteoporosis after the artificial joint operation.
本研究旨在探讨唑来膦酸（zoledronate，ZOL）对钛（Ti）颗粒诱导的破骨细胞前体细胞 RAW264.7 分化的影响，探索 ZOL 预防和治疗人工关节置换术后假体周围骨质疏松症的可能性。
体外培养 RAW264.7 细胞，制备 Ti 颗粒，采用 MTT 法绘制 RAW264.7 细胞增殖曲线，寻找 ZOL 最佳干预浓度。将细胞分为 3 组：对照组、Ti 颗粒组、Ti 颗粒+ZOL 组。采用酒石酸抗酸性磷酸酶（tartaric acid-resistant acid phosphatase，TRAP）染色观察细胞形态，采用生化法检测细胞上清液中 TRAP 的活性，采用甲苯胺蓝染色法检测骨吸收陷窝数，采用实时聚合酶链反应（polymerase chain reaction，PCR）检测 、 、 、 基因的表达，采用 Western blot 分析检测核因子 κB 受体活化因子配体（receptor activator of nuclear factor kappa-B ligand，RANK）、活化 T 细胞核因子 1（nuclear factor of activated T cells cytoplasmic 1，NFATcl）和基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）的蛋白表达。
Ti 颗粒可刺激 RAW264.7 细胞向破骨细胞分化，增加 TRAP 活性、骨吸收陷窝数以及 RANK、NFATcl 和 MMP-9 基因和蛋白的表达，而 ZOL 则可抑制 Ti 颗粒对 RAW264.7 细胞向破骨细胞分化的作用。
ZOL 可有效抑制 Ti 颗粒诱导的 RAW264.7 细胞向破骨细胞分化，降低 TRAP 活性，减少骨吸收陷窝数，降低 RANK、NFATcl 和 MMP-9 基因和蛋白的表达，有望成为人工关节置换术后预防和治疗假体周围骨质疏松症的候选药物。
