Effect of BioAggregate on Receptor Activator of Nuclear Factor-Kappa B Ligand-induced Osteoclastogenesis from Murine Macrophage Cell Line In Vitro
Effect of BioAggregate on Receptor Activator of Nuclear Factor-Kappa B Ligand-induced Osteoclastogenesis from Murine Macrophage Cell Line In Vitro
复制标题
BioAggregate 对核因子-κ B 配体受体激活剂诱导的小鼠巨噬细胞系体外破骨细胞生成的影响
DOI：
10.1016/j.joen.2015.03.021
复制
发表时间：
2015-08-01
影响因子：
4.2
通讯作者：
Peng, Bin
中科院分区：
文献类型：
--
作者：
Zhang, Jie;Zhu, Lingxin;Peng, Bin
简介：本研究旨在探讨BioAggregate（一种基于硅酸钙的纳米颗粒生物陶瓷）在体外对核因子κ B受体激活因子配体（RANKL）诱导的破骨细胞分化和骨吸收的调节作用，并阐明其分子机制。BioAggregate的性能与Pro Root矿物质三氧化物聚集体（MTA）的性能相当。方法：小鼠巨噬细胞系RAW 264.7的细胞用不同浓度的BioAggregate和MTA提取物处理。采用细胞计数试剂盒-8测定法评价材料浸提液的细胞毒性。RANKL诱导的破骨细胞分化和功能通过抗酒石酸酸性磷酸酶染色、F-肌动蛋白染色和陷窝吸收陷窝测定进行评估。实时定量聚合酶链反应检测破骨细胞功能相关mRNA表达。Western blot和免疫荧光法检测相关分子信号通路。结果：BioAggregate浸提液剂量依赖性地抑制RANKL诱导的破骨细胞形成和吸收能力，无明显细胞毒性。暴露于BioAggregate浸提液的RAW 264.7细胞也显示破骨细胞相关基因和转录因子的RANKL刺激mRNA表达降低。此外，BioAggregate提取物处理的细胞表现出TRAF 6表达减弱，丝裂原活化蛋白激酶信号转导抑制，以及在RANKL存在下NFATc 1和c-Fos核转位减少。MTA诱导的效应相当。结论：BioAggregate和MTA在体外对破骨细胞的分化和功能表现出相当的抑制作用，我们的研究结果为生物陶瓷介导的抗破骨细胞生成活性的机制提供了有价值的见解。
Introduction: This study aimed to investigate the effect of BioAggregate, a calcium silicate-based nanoparticulate bioceramic, on the regulation of receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclast differentiation and bone resorption in vitro, as well as to delineate the underlying molecular mechanism. The performance of BioAggregate was corn-pared with that of Pro Root mineral trioxide aggregate (MTA). Methods: Cells of a murine macrophage cell line RAW 264.7 were treated with various concentrations of BioAggregate and MTA extracts. Cytotoxicity of material extracts was evaluated with Cell Counting Kit-8 assay. RANKL-induced osteoclast differentiation and function were assessed with tartrate-resistant acid phosphatase staining, F-actin staining, and lacunar resorption pits assay. The mRNA expression associated with osteoclast function was detected with quantitative real-time polymerase chain reaction. Related molecular signaling pathways were investigated with Western blot and immunofluorescence. Results: BioAggregate extracts dose-dependently inhibited RANKL-induced osteoclast formation and resorption capacity without evident cytotoxicity. RAW 264.7 cells exposed to BioAggregate extracts also presented a decrease in RANKL-stimulated mRNA expression of osteoclast-related genes and transcription factors. Moreover, cells treated with BioAggregate extracts exhibited attenuated TRAF6 expression, suppressed mitogen-activated protein kinase signaling, and decreased nuclear translocation of NFATc1 and c-Fos in the presence of RANKL. Comparable effects were induced by MTA. Conclusions: BioAggregate and MTA exhibit comparable inhibitory effect on osteoclast differentiation and function in vitro, and our findings provide valuable insights into the mechanism of bioceramic-mediated anti-osteoclastogenic activity.