Zhang Jie, Zhu Lingxin, Yan Ping, Peng Bin
State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
J Endod. 2015 Aug;41(8):1265-71. doi: 10.1016/j.joen.2015.03.021. Epub 2015 May 12.
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 compared with that of ProRoot mineral trioxide aggregate (MTA).
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.
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.
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.
本研究旨在探讨基于硅酸钙的纳米颗粒生物陶瓷BioAggregate对体外核因子κB受体激活剂配体（RANKL）诱导的破骨细胞分化和骨吸收的调节作用，并阐明其潜在的分子机制。将BioAggregate的性能与ProRoot三氧化矿物凝聚体（MTA）进行比较。
用不同浓度的BioAggregate和MTA提取物处理小鼠巨噬细胞系RAW 264.7细胞。用细胞计数试剂盒-8法评估材料提取物的细胞毒性。用抗酒石酸酸性磷酸酶染色、F-肌动蛋白染色和陷窝吸收凹坑试验评估RANKL诱导的破骨细胞分化和功能。用定量实时聚合酶链反应检测与破骨细胞功能相关的mRNA表达。用蛋白质免疫印迹法和免疫荧光法研究相关分子信号通路。
BioAggregate提取物呈剂量依赖性抑制RANKL诱导的破骨细胞形成和吸收能力，且无明显细胞毒性。暴露于BioAggregate提取物的RAW 264.7细胞还表现出RANKL刺激的破骨细胞相关基因和转录因子的mRNA表达降低。此外，在存在RANKL的情况下，用BioAggregate提取物处理的细胞表现出TRAF6表达减弱、丝裂原活化蛋白激酶信号传导受抑制以及NFATc1和c-Fos的核转位减少。MTA也诱导了类似的效果。
BioAggregate和MTA在体外对破骨细胞分化和功能表现出类似的抑制作用，我们的研究结果为生物陶瓷介导的抗破骨细胞活性机制提供了有价值的见解。
