在原代大鼠肺泡巨噬细胞和正常大鼠间皮细胞 NRM2 中体外筛选矿棉毒理学的方法步骤

IF 4.8 2区 医学 Q1 TOXICOLOGY
Christina Ziemann, Florian Schulz, Christoph Koch, Mette Solvang, Annette Bitsch
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引用次数: 0

摘要

人造玻璃纤维(MMVF)包括各种用于隔热和隔音的材料,其中包括岩棉。根据尺寸、耐久性和剂量的不同,人造玻璃纤维可能会对健康产生不良影响。因此,我们非常需要对新型 MMVF 进行早期预测性体外(基因)毒性筛选,以确保接触这些材料的工人和消费者的安全。在此,我们以原代大鼠肺泡巨噬细胞(AM)和正常大鼠间皮细胞(NRM2)为起点,研究了体外筛选的关键决定因素和陷阱。石羊毛纤维(RIF56008)作为示范性 MMVF(纤维状与磨碎状,以估计纤维形状的影响),长铁石棉(石棉)作为不溶性纤维参考。根据不同的方法(从低负荷到假定超负荷:0.5、5 和 50 µg/cm2),对材料进行了全面的特征描述,并确定了体内相关的体外浓度。培养 4-48 小时后,分析了某些读数,并通过光和荧光耦合暗视野显微镜研究了材料吸收情况。DNA 链断裂诱导与形态无关,在两种细胞中几乎都不存在。然而,NRM2 表现出了与材料、形态和浓度相关的膜损伤、CINC-1 释放、细胞数量减少以及双核细胞诱导(石棉> RIF56008 >RIF56008地面)。与 NRM2 相反,石棉在 AM 中几乎没有活性,CINC-1 的释放仅由 RIF56008 诱导。总之,要确定一种与 MMVF 相适应的、具有预测性的体外(基因)毒性筛选工具,应根据所选细胞模型的体内相关性、敏感性和特异性,仔细选择参考点、终点和浓度。接下来,应评估进一步的终点,最好能通过体内数据验证其预测性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Methodological steps forward in toxicological in vitro screening of mineral wools in primary rat alveolar macrophages and normal rat mesothelial NRM2 cells

Methodological steps forward in toxicological in vitro screening of mineral wools in primary rat alveolar macrophages and normal rat mesothelial NRM2 cells

Man-made vitreous fibers (MMVF) comprise diverse materials for thermal and acoustic insulation, including stone wool. Depending on dimension, durability, and dose, MMVF might induce adverse health effects. Therefore, early predictive in vitro (geno)toxicity screening of new MMVF is highly desired to ensure safety for exposed workers and consumers. Here, we investigated, as a starting point, critical in vitro screening determinants and pitfalls using primary rat alveolar macrophages (AM) and normal rat mesothelial cells (NRM2). A stone wool fiber (RIF56008) served as an exemplary MMVF (fibrous vs. ground to estimate impact of fiber shape) and long amosite (asbestos) as insoluble fiber reference. Materials were comprehensively characterized, and in vivo-relevant in vitro concentrations defined, based on different approaches (low to supposed overload: 0.5, 5 and 50 µg/cm2). After 4–48 h of incubation, certain readouts were analyzed and material uptake was investigated by light and fluorescence-coupled darkfield microscopy. DNA-strand break induction was not morphology-dependent and nearly absent in both cell types. However, NRM2 demonstrated material-, morphology- and concentration-dependent membrane damage, CINC-1 release, reduction in cell count, and induction of binucleated cells (asbestos > RIF56008 > RIF56008 ground). In contrast to NRM2, asbestos was nearly inactive in AM, with CINC-1 release solely induced by RIF56008. In conclusion, to define an MMVF-adapted, predictive in vitro (geno)toxicity screening tool, references, endpoints, and concentrations should be carefully chosen, based on in vivo relevance, and sensitivity and specificity of the chosen cell model. Next, further endpoints should be evaluated, ideally with validation by in vivo data regarding their predictivity.

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来源期刊
Archives of Toxicology
Archives of Toxicology 医学-毒理学
CiteScore
11.60
自引率
4.90%
发文量
218
审稿时长
1.5 months
期刊介绍: Archives of Toxicology provides up-to-date information on the latest advances in toxicology. The journal places particular emphasis on studies relating to defined effects of chemicals and mechanisms of toxicity, including toxic activities at the molecular level, in humans and experimental animals. Coverage includes new insights into analysis and toxicokinetics and into forensic toxicology. Review articles of general interest to toxicologists are an additional important feature of the journal.
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