Crystalline structure of pulverized dental calculus induces cell death in oral epithelial cells
Crystalline structure of pulverized dental calculus induces cell death in oral epithelial cells
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粉碎牙石的晶体结构诱导口腔上皮细胞细胞死亡
DOI：
10.1111/jre.12520
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发表时间：
2018
影响因子：
3.5
通讯作者：
Hara Y
中科院分区：
文献类型：
--
作者：
Ziauddin S M;Yoshimura A;Montenegro Raudales J L;Ozaki Y;Higuchi K;Ukai T;Kaneko T;Miyazaki T;Latz E;Hara Y
牙结石是附着在牙齿表面的矿化存款。我们已经表明，牙结石的细胞摄取触发了核苷酸结合寡聚化结构域样受体家族含吡林结构域3（NLRP 3）炎性体激活，导致白细胞介素1β前体在小鼠和人类吞噬细胞中加工成成熟形式。NLRP 3炎性体的激活也诱导了这些细胞中程序性细胞死亡的裂解形式，即细胞凋亡。然而，牙结石对牙周组织中其他细胞类型的影响尚未研究。本研究的目的是确定牙结石是否能诱导口腔上皮细胞的细胞死亡。材料和方法将HSC-2人口腔鳞癌细胞、HOMK 107人原代口腔上皮细胞和永生化小鼠巨噬细胞暴露于牙结石或其成分之一羟基磷灰石晶体中。对于抑制测定，在存在或不存在细胞松弛素D（内吞作用抑制剂）、z-YVAD-fastrin（半胱天冬酶-1抑制剂）或格列本脲（NLRP 3炎性体抑制剂）的情况下将细胞暴露于牙石。通过测量乳酸脱氢酶（LDH）释放和碘化丙啶染色测定细胞毒性。通过酶联免疫吸附试验定量肿瘤坏死因子-α的产生。口腔上皮屏障功能的渗透性assay.ResultsDental结石诱导HSC-2细胞的细胞死亡，判断LDH释放和碘化丙啶染色。牙结石也诱导HOMK 107细胞释放LDH。热处理后，牙结石失去了在小鼠巨噬细胞中诱导肿瘤坏死因子-α的能力，但可以诱导HSC-2细胞释放LDH，表明无机成分在细胞死亡中起主要作用。羟基磷灰石晶体还诱导HSC-2和HOMK 107细胞中的细胞死亡，如通过LDH释放所判断的，表明晶体颗粒诱导细胞死亡的能力。牙结石诱导的细胞死亡被细胞松弛素D、z-YVAD-favorite和格列本脲显著抑制，表明NLRP 3炎性小体参与。在渗透性测定，牙结石衰减的屏障功能的HSC-2细胞monolays.ConclusionDental结石诱导pyroptotic细胞死亡在人类口腔上皮细胞和晶体结构在这一过程中起着重要作用。牙结石引起的口腔上皮细胞死亡可能是牙周炎的重要病因。
Background and ObjectiveDental calculus is a mineralized deposit attached to the tooth surface. We have shown that cellular uptake of dental calculus triggers nucleotide‐binding oligomerization domain‐like receptor family pyrin domain‐containing 3 (NLRP3) inflammasome activation, leading to the processing of the interleukin‐1β precursor into its mature form in mouse and human phagocytes. The activation of the NLRP3 inflammasome also induced a lytic form of programmed cell death, pyroptosis, in these cells. However, the effects of dental calculus on other cell types in periodontal tissue have not been investigated. The aim of this study was to determine whether dental calculus can induce cell death in oral epithelial cells.Material and MethodsHSC‐2 human oral squamous carcinoma cells, HOMK107 human primary oral epithelial cells and immortalized mouse macrophages were exposed to dental calculus or 1 of its components, hydroxyapatite crystals. For inhibition assays, the cells were exposed to dental calculus in the presence or absence of cytochalasin D (endocytosis inhibitor), z‐YVAD‐fmk (caspase‐1 inhibitor) or glyburide (NLRP3 inflammasome inhibitor). Cytotoxicity was determined by measuring lactate dehydrogenase (LDH) release and staining with propidium iodide. Tumor necrosis factor‐α production was quantified by enzyme‐linked immunosorbent assay. Oral epithelial barrier function was examined by permeability assay.ResultsDental calculus induced cell death in HSC‐2 cells, as judged by LDH release and propidium iodide staining. Dental calculus also induced LDH release from HOMK107 cells. Following heat treatment, dental calculus lost its capacity to induce tumor necrosis factor‐α in mouse macrophages, but could induce LDH release in HSC‐2 cells, indicating a major role of inorganic components in cell death. Hydroxyapatite crystals also induced cell death in both HSC‐2 and HOMK107 cells, as judged by LDH release, indicating the capacity of crystal particles to induce cell death. Cell death induced by dental calculus was significantly inhibited by cytochalasin D, z‐YVAD‐fmk and glyburide, indicating NLRP3 inflammasome involvement. In permeability assays, dental calculus attenuated the barrier function of HSC‐2 cell monolayers.ConclusionDental calculus induces pyroptotic cell death in human oral epithelial cells and the crystalline structure plays a major role in this process. Oral epithelial cell death induced by dental calculus might be important for the etiology of periodontitis.
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1999
期刊：
Australian endodontic journal : the journal of the Australian Society of Endodontology Inc
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