Table_1_Morphological and lipid metabolism alterations in macrophages exposed to model environmental nanoplastics traced by high-resolution synchrotron techniques.docx

The release of nanoplastics (NPs) in the environment is a significant health concern for long-term exposed humans. Although their usage has certainly revolutionized several application fields, at nanometer size, NPs can easily interact at the cellular level, resulting in potential harmful effects. Micro/Nanoplastics (M/NPs) have a demonstrated impact on mammalian endocrine components, such as the thyroid, adrenal gland, testes, and ovaries, while more investigations on prenatal and postnatal exposure are urgently required. The number of literature studies on the NPs’ presence in biological samples is increasing. However, only a few offer a close study on the model environmental NP–immune system interaction exploited by advanced microscopy techniques. The present study highlights substantial morphological and lipid metabolism alterations in human M1 macrophages exposed to labeled polypropylene and polyvinyl chloride nanoparticles (PP and PVC NPs) (20 μg/ml). The results are interpreted by advanced microscopy techniques combined with standard laboratory tests and fluorescence microscopy. We report the accurate detection of polymeric nanoparticles doped with cadmium selenide quantum dots (CdSe-QDs NPs) by following the Se (L line) X-ray fluorescence emission peak at higher sub-cellular resolution, compared to the supportive light fluorescence microscopy. In addition, scanning transmission X-ray microscopy (STXM) imaging successfully revealed morphological changes in NP-exposed macrophages, providing input for Fourier transform infrared (FTIR) spectroscopy analyses, which underlined the chemical modifications in macromolecular components, specifically in lipid response. The present evidence was confirmed by quantifying the lipid droplet (LD) contents in PP and PVC NPs-exposed macrophages (0–100 μg/ml) by Oil Red O staining. Hence, even at experimental NPs' concentrations and incubation time, they do not significantly affect cell viability; they cause an evident lipid metabolism impairment, a hallmark of phagocytosis and oxidative stress.

纳米塑料（NPs）向环境中的释放，对长期暴露的人类构成了重大的健康隐患。尽管其应用确实在多个领域引发了革命性的变化，但在纳米尺度上，NPs能够轻易地在细胞层面上发生相互作用，从而产生潜在的有害影响。微/纳米塑料（M/NPs）已被证实对哺乳动物的内分泌成分，如甲状腺、肾上腺、睾丸和卵巢，具有影响，而关于孕前和产后暴露的更多研究亟待进行。关于NPs在生物样本中存在的文献研究数量正在增加。然而，仅有少数研究通过高级显微镜技术对模型环境中的NP-免疫系统相互作用进行了深入研究。本研究突出了人类M1巨噬细胞在标记的多聚丙烯和聚氯乙烯纳米颗粒（PP和PVC NPs）（20 μg/ml）暴露下所发生的显著形态学和脂质代谢改变。结果通过结合高级显微镜技术和标准实验室测试以及荧光显微镜进行解读。我们通过在更高亚细胞分辨率的硒（L线）X射线荧光发射峰处跟踪，精确检测了掺杂硒化镉量子点（CdSe-QDs NPs）的聚合物纳米颗粒，与辅助性的光学荧光显微镜相比。此外，扫描透射X射线显微镜（STXM）成像成功揭示了NP暴露巨噬细胞的形态学变化，为傅里叶变换红外（FTIR）光谱分析提供了输入，强调了在大分子成分，尤其是脂质反应中的化学修饰。通过油红O染色定量PP和PVC NPs暴露巨噬细胞（0–100 μg/ml）中的脂滴（LD）含量，本研究证据得到了证实。因此，即使在实验性NPs的浓度和孵育时间内，它们也不会显著影响细胞活力；它们会导致明显的脂质代谢障碍，这是吞噬作用和氧化应激的特征。