Ferroptosis is involved in trophoblast cells cytotoxicity induced by black phosphorus nanoparticles

Ruolan You; Yaming Mu; Jiaqi Zhou; Chunying Wang; Zhenya Fang; Yu Liu; Shiyu Liu; Qingfeng Zhai; Changqing Zhang

doi:10.1016/j.tox.2024.153810

Ferroptosis is involved in trophoblast cells cytotoxicity induced by black phosphorus nanoparticles

Toxicology. 2024 Apr 21:505:153810. doi: 10.1016/j.tox.2024.153810. Online ahead of print.

Authors

Ruolan You¹, Yaming Mu¹, Jiaqi Zhou¹, Chunying Wang², Zhenya Fang², Yu Liu², Shiyu Liu³, Qingfeng Zhai⁴, Changqing Zhang⁵

Affiliations

¹ School of Public Health, Shandong Second Medical University, Weifang 261053, China.
² Key Laboratory of Maternal & Fetal Medicine of National Health Commission of China，Shandong Provincial Maternal and Child Health Care Hospital Affiliated to Qingdao University, Jinan 250014, China.
³ International Center, Jinan Foreign Language School, Jinan 250108, China.
⁴ School of Public Health, Shandong Second Medical University, Weifang 261053, China. Electronic address: sdzzqqff@163.com.
⁵ Key Laboratory of Maternal & Fetal Medicine of National Health Commission of China，Shandong Provincial Maternal and Child Health Care Hospital Affiliated to Qingdao University, Jinan 250014, China. Electronic address: zhan0396@e.ntu.edu.sg.

PMID: 38653377
DOI: 10.1016/j.tox.2024.153810

Abstract

Black phosphorus (BP) is a new type of nanomaterial, which has been widely used in many biomedical fields due to its superior properties, but there are few studies on the toxicity of BP, especially in the reproductive system. To explore the effects of BP exposure on reproduction and reveal its molecular mechanism, we firstly investigated the potential toxicity of black phosphorus nanoparticles (BPNPs) in vivo. The results showed that BP exposure in pregnant mice can reduce the weight of fetal mice and placenta. H&E staining further indicated the changes of placental cross-section and vascular remodeling after BP treatment. Then, human exvillous trophoblast HTR8/SVneo was treated with different concentrations of BPNPs. We found that BPNPs induced significant cytotoxicity, including dose-dependent reduction of cell viability and proliferation. Trophoblast cell migration and invasion were also impaired by BPNPs exposure. Moreover, pretreatment with Cytochalasin D (Cyto-D), a classical phagocytic inhibitor, alleviated the decline of cell viability induced by BPNPs. Transcriptome sequencing showed that BPNPs exposure led to ferroptosis. Subsequently, the related indexes of ferroptosis were detected, including increase of iron ion concentration, decrease of the ferroptosis marker, GPX4 (Glutathione Peroxidase 4), increase of FTL (Ferritin Light Chain), and increase of lipid peroxidation indexes (MDA level and decrease of GSH level). In addition, ferroptosis inhibitors (Fer-1 and DFO) pretreatment can alleviate both the cytotoxic effects and functional impairment induced by BPNPs. In summary, our study confirmed the reproductive toxicity of BPNPs for the first time, and constructed BPNPs injury model in vitro using human villus trophoblast cells and revealed the role of ferroptosis in this process, which deepened our understanding of the biosafety of black phosphorus nanomaterials.

Keywords: Black phosphorus; Cytotoxicity; Ferroptosis; Trophoblast.