利用高通量筛选方法对焚化前和焚化后的有机改性纳米粘土对巨噬细胞的体外炎症和毒性进行评估。

IF 7.2 1区 医学 Q1 TOXICOLOGY
Todd A Stueckle, Jake Jensen, Jayme P Coyle, Raymond Derk, Alixandra Wagner, Cerasela Zoica Dinu, Tiffany G Kornberg, Sherri A Friend, Alan Dozier, Sushant Agarwal, Rakesh K Gupta, Liying W Rojanasakul
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引用次数: 0

摘要

背景:有机改性纳米粘土(ONC)是一种带有有机涂层的二维蒙脱石,越来越多地用于改善纳米复合材料的性能。然而,人们对纳米粘土生命周期中的肺部健康风险知之甚少,即使有越来越多的证据表明在职业环境中会接触到空气中的微粒。最近,小鼠口咽部吸入焚化前和焚化后的 ONC 会引起低度、持续性肺部炎症,并伴有促纤维化信号反应,其作用模式尚不清楚。我们假设,纳米瓷的有机涂层和焚烧状态决定了巨噬细胞的炎症细胞因子秘书特征和细胞毒性反应。为了验证这一假设,我们将分化的人类巨噬细胞(THP-1)急性暴露于(0-20 µg/cm2)未涂层的原始纳米粘土(CloisNa)、ONC(Clois30B)、它们的焚烧副产品(I-CloisNa 和 I-Clois30B)以及结晶二氧化硅(CS),然后进行细胞毒性和炎症终点检测。巨噬细胞同时暴露于脂多糖(LPS)或不含 LPS 的培养基中,以评估 NF-κB 通路在巨噬细胞对纳米土处理的反应中的作用。将这些数据与雄性C57Bl/6J小鼠吸入30微克和300微克相同颗粒后的炎症反应进行比较:结果:与CloisNa和未暴露的对照组相比,在不含LPS的培养基中,CloisNa暴露会导致线粒体去极化,而Clois30B暴露会导致巨噬细胞存活率降低、细胞毒性增强以及损伤相关分子模式(IL-1α和ATP)的显著释放。低剂量 CloisNa 引发的 LPS 会导致酪蛋白酶 B/Caspage-1/IL-1β 释放增加,而高剂量则会导致细胞凋亡。暴露于 Clois30B 会导致剂量依赖性的 THP-1 细胞脓毒症,表现为酪蛋白酶 B 和 IL-1β 的释放以及 Gasdermin D 的裂解。焚烧消除了 Clois30B 的细胞毒性和炎症效应,而 I-CloisNa 仍保留了一些轻微的炎症潜能。比较分析表明,体外巨噬细胞活力、炎症小体终点和促炎细胞因子谱与小鼠支气管肺泡灌洗液的炎症指标(包括炎症细胞招募)显著相关:结论:有机涂层的存在和焚烧状态会影响暴露于人类巨噬细胞后的炎症和细胞毒性反应。带有季铵牛脂涂层的 Clois30B 可诱导强烈的细胞膜损伤和热昏迷效应,这种效应在焚烧后被消除。相反,焚烧后的纳米粘土暴露主要导致 THP-1 细胞释放更多的炎症细胞因子。总之,焚烧前的纳米黏土与巨噬细胞膜成分相互作用(分子启动事件),增加了促炎介质,并增加了肺纤维化不良后果途径中的炎症细胞招募(两个关键事件)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
In vitro inflammation and toxicity assessment of pre- and post-incinerated organomodified nanoclays to macrophages using high-throughput screening approaches.

Background: Organomodified nanoclays (ONC), two-dimensional montmorillonite with organic coatings, are increasingly used to improve nanocomposite properties. However, little is known about pulmonary health risks along the nanoclay life cycle even with increased evidence of airborne particulate exposures in occupational environments. Recently, oropharyngeal aspiration exposure to pre- and post-incinerated ONC in mice caused low grade, persistent lung inflammation with a pro-fibrotic signaling response with unknown mode(s) of action. We hypothesized that the organic coating presence and incineration status of nanoclays determine the inflammatory cytokine secretary profile and cytotoxic response of macrophages. To test this hypothesis differentiated human macrophages (THP-1) were acutely exposed (0-20 µg/cm2) to pristine, uncoated nanoclay (CloisNa), an ONC (Clois30B), their incinerated byproducts (I-CloisNa and I-Clois30B), and crystalline silica (CS) followed by cytotoxicity and inflammatory endpoints. Macrophages were co-exposed to lipopolysaccharide (LPS) or LPS-free medium to assess the role of priming the NF-κB pathway in macrophage response to nanoclay treatment. Data were compared to inflammatory responses in male C57Bl/6J mice following 30 and 300 µg/mouse aspiration exposure to the same particles.

Results: In LPS-free media, CloisNa exposure caused mitochondrial depolarization while Clois30B exposure caused reduced macrophage viability, greater cytotoxicity, and significant damage-associated molecular patterns (IL-1α and ATP) release compared to CloisNa and unexposed controls. LPS priming with low CloisNa doses caused elevated cathepsin B/Caspage-1/IL-1β release while higher doses resulted in apoptosis. Clois30B exposure caused dose-dependent THP-1 cell pyroptosis evidenced by Cathepsin B and IL-1β release and Gasdermin D cleavage. Incineration ablated the cytotoxic and inflammatory effects of Clois30B while I-CloisNa still retained some mild inflammatory potential. Comparative analyses suggested that in vitro macrophage cell viability, inflammasome endpoints, and pro-inflammatory cytokine profiles significantly correlated to mouse bronchioalveolar lavage inflammation metrics including inflammatory cell recruitment.

Conclusions: Presence of organic coating and incineration status influenced inflammatory and cytotoxic responses following exposure to human macrophages. Clois30B, with a quaternary ammonium tallow coating, induced a robust cell membrane damage and pyroptosis effect which was eliminated after incineration. Conversely, incinerated nanoclay exposure primarily caused elevated inflammatory cytokine release from THP-1 cells. Collectively, pre-incinerated nanoclay displayed interaction with macrophage membrane components (molecular initiating event), increased pro-inflammatory mediators, and increased inflammatory cell recruitment (two key events) in the lung fibrosis adverse outcome pathway.

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来源期刊
CiteScore
15.90
自引率
4.00%
发文量
69
审稿时长
6 months
期刊介绍: Particle and Fibre Toxicology is an online journal that is open access and peer-reviewed. It covers a range of disciplines such as material science, biomaterials, and nanomedicine, focusing on the toxicological effects of particles and fibres. The journal serves as a platform for scientific debate and communication among toxicologists and scientists from different fields who work with particle and fibre materials. The main objective of the journal is to deepen our understanding of the physico-chemical properties of particles, their potential for human exposure, and the resulting biological effects. It also addresses regulatory issues related to particle exposure in workplaces and the general environment. Moreover, the journal recognizes that there are various situations where particles can pose a toxicological threat, such as the use of old materials in new applications or the introduction of new materials altogether. By encompassing all these disciplines, Particle and Fibre Toxicology provides a comprehensive source for research in this field.
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