Nanoparticulate bioceramic putty suppresses osteoclastogenesis and inflammatory bone loss in mice via inhibition of TRAF6-mediated signalling pathways: A laboratory investigation

IF 5.4 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

International endodontic journal Pub Date : 2024-02-25 DOI:10.1111/iej.14051

Zijun Wang, Jie Zhang, Xiaoyue Sun, Jingjing Yu, Bingqian Liu, Bin Peng, Li Wang, Jingwen Yang, Lingxin Zhu

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引用次数: 0

Abstract

Aim

This study aimed to determine the effects of iRoot BP Plus on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis in vitro and inflammation-mediated bone resorption in vivo and investigated the underlying molecular mechanisms.

Methodology

CCK-8 was performed to test cell viability in RANKL-induced RAW 264.7 cells and BMDMs in response to iRoot BP Plus. The effect of iRoot BP Plus on osteoclastogenesis was determined using TRAP staining and phalloidin staining, respectively. Pit formation assay was conducted to measure osteoclast resorptive capacity. Western blot and qPCR were performed to examine osteoclast-related proteins and gene expression, respectively. Western blot was also used to investigate the signalling pathways involved. For in vivo experiments, an LPS-induced mouse calvarial bone resorption model was established to analyse the effect of iRoot BP Plus on bone resorption (n = 6 per group). At 7 days, mouse calvaria were collected and prepared for histological analysis.

Results

We identified that iRoot BP Plus extracts significantly attenuated RANKL-induced osteoclastogenesis, reduced sealing zone formation, restrained osteolytic capacity and decreased osteoclast-specific gene expression (p < .01). Mechanistically, iRoot BP Plus extracts reduced TRAF6 via proteasomal degradation, then suppressed the phosphorylation of mitogen-activated protein kinases (MAPKs), blocked the nuclear translocation of c-Fos and diminished nuclear factor-κB (NF-κB) p65 and NFATc1 accumulation. Consistent with the in vitro results, iRoot BP Plus extracts attenuated osteoclast activity thus protecting against inflammatory bone resorption in vivo (p < .05), which was accompanied by a suppression of TRAF6, c-Fos, NFATc1 and cathepsin K expression.

Conclusion

These findings provide valuable insights into the signalling mechanisms underlying nanoparticulate bioceramic putty-mediated bone homeostasis.

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纳米微粒生物陶瓷腻子通过抑制 TRAF6 介导的信号通路抑制小鼠破骨细胞生成和炎性骨质流失：实验室研究

目的：本研究旨在确定iRoot BP Plus对体外核因子κB配体受体激活剂（RANKL）诱导的破骨细胞生成和体内炎症介导的骨吸收的影响，并研究其潜在的分子机制：用 CCK-8 检测 RANKL 诱导的 RAW 264.7 细胞和 BMDMs 对 iRoot BP Plus 的反应。iRoot BP Plus 对破骨细胞生成的影响分别通过 TRAP 染色和类磷脂染色来确定。坑形成试验用于测量破骨细胞的吸收能力。Western 印迹和 qPCR 分别用于检测破骨细胞相关蛋白和基因表达。此外，还使用 Western 印迹法研究了相关的信号通路。在体内实验中，建立了 LPS 诱导的小鼠小腿骨吸收模型，以分析 iRoot BP Plus 对骨吸收的影响（每组 6 人）。7 天后，收集小鼠钙片并准备进行组织学分析：这些发现为了解纳米微粒生物陶瓷腻子介导骨平衡的信号机制提供了宝贵的见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International endodontic journal 医学-牙科与口腔外科

CiteScore

10.20

自引率

28.00%

发文量

195

审稿时长

4-8 weeks

期刊介绍： The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.