Department of General Dentistry, the Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing, Jiangsu, China.
Jiangsu Province Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, Jiangsu, China.
Int Endod J. 2024 Jun;57(6):727-744. doi: 10.1111/iej.14055. Epub 2024 Mar 4.
This study aimed to investigate the anti-inflammatory and odontoblastic effects of cerium-containing mesoporous bioactive glass nanoparticles (Ce-MBGNs) on dental pulp cells as novel pulp-capping agents.
Ce-MBGNs were synthesized using a post-impregnation strategy based on the antioxidant properties of Ce ions and proposed the first use of Ce-MBGNs for pulp-capping application. The biocompatibility of Ce-MBGNs was analysed using the CCK-8 assay and apoptosis detection. Additionally, the reactive oxygen species (ROS) scavenging ability of Ce-MBGNs was measured using the 2,7-Dichlorofuorescin Diacetate (DCFH-DA) probe. The anti-inflammatory effect of Ce-MBGNs on THP-1 cells was further investigated using flow cytometry and quantitative real-time polymerase chain reaction (RT-qPCR). Moreover, the effect of Ce-MBGNs on the odontoblastic differentiation of the dental pulp cells (DPCs) was assessed by combined scratch assays, RT-qPCR, western blotting, immunocytochemistry, Alizarin Red S staining and tissue-nonspecific alkaline phosphatase staining. Analytically, the secretions of tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected with enzyme-linked immunosorbent assay (ELISA).
Ce-MBGNs were confirmed to effectively scavenge ROS in THP-1-derived macrophages and DPCs. Flow cytometry and RT-qPCR assays revealed that Ce-MBGNs significantly inhibited the M1 polarization of macrophages (Mφ). Furthermore, the protein levels of TNF-α and IL-1β were downregulated in THP-1-derived macrophages after stimulation with Ce-MBGNs. With a step-forward virtue of promoting the odontoblastic differentiation of DPCs, we further confirmed that Ce-MBGNs could regulate the formation of a conductive immune microenvironment with respect to tissue repair in DPCs, which was mediated by macrophages.
Ce-MBGNs protected cells from self-produced oxidative damage and exhibited excellent immunomodulatory and odontoblastic differentiation effects on DPCs. As a pulp-capping agent, this novel biomaterial can exert anti-inflammatory effects and promote restorative dentine regeneration in clinical treatment. We believe that this study will stimulate further correlative research on the development of advanced pulp-capping agents.
本研究旨在探讨含铈介孔生物活性玻璃纳米颗粒(Ce-MBGNs)作为新型盖髓剂对牙髓细胞的抗炎和成牙本质作用。
采用基于铈离子抗氧化特性的后浸渍策略合成 Ce-MBGNs,并首次提出将 Ce-MBGNs 用于盖髓应用。通过 CCK-8 测定和凋亡检测分析 Ce-MBGNs 的生物相容性。此外,还使用 2,7-二氯荧光素二乙酸酯(DCFH-DA)探针测量 Ce-MBGNs 的清除活性氧(ROS)的能力。通过流式细胞术和实时定量聚合酶链反应(RT-qPCR)进一步研究 Ce-MBGNs 对 THP-1 细胞的抗炎作用。此外,通过划痕实验、RT-qPCR、Western blot、免疫细胞化学、茜素红 S 染色和组织非特异性碱性磷酸酶染色评估 Ce-MBGNs 对牙髓细胞(DPCs)成牙本质分化的影响。通过酶联免疫吸附测定(ELISA)检测肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的分泌。
Ce-MBGNs 被证实可有效清除 THP-1 衍生巨噬细胞和 DPCs 中的 ROS。流式细胞术和 RT-qPCR 分析显示,Ce-MBGNs 可显著抑制巨噬细胞(Mφ)的 M1 极化。此外,Ce-MBGNs 刺激后 THP-1 衍生巨噬细胞中 TNF-α和 IL-1β的蛋白水平下调。更进一步,我们还证实 Ce-MBGNs 可促进 DPCs 的成牙本质分化,从而调节与 DPCs 组织修复相关的具有传导性的免疫微环境,这是由巨噬细胞介导的。
Ce-MBGNs 可保护细胞免受自身产生的氧化损伤,并对 DPCs 表现出优异的免疫调节和成牙本质分化作用。作为一种盖髓剂,这种新型生物材料在临床治疗中具有抗炎作用,并可促进修复性牙本质再生。我们相信,这项研究将激发关于先进盖髓剂开发的进一步相关研究。