NiONP-Induced Oxidative Stress and Mitochondrial Impairment in an In Vitro Pulmonary Vascular Cell Model Mimicking Endothelial Dysfunction

Abstract

The development and use of nanomaterials, especially of nickel oxide nanoparticles (NiONPs), is expected to provide many benefits but also has raised concerns about the potential human health risks. Inhaled NPs are known to exert deleterious cardiovascular side effects, including pulmonary hypertension. Consequently, patients with pulmonary hypertension (PH) could be at increased risk for morbidity. The objective of this study was to compare the toxic effects of NiONPs on human pulmonary artery endothelial cells (HPAEC) under physiological and pathological conditions. The study was conducted with an in vitro model mimicking the endothelial dysfunction observed in PH. HPAEC were cultured under physiological (static and normoxic) or pathological (20% cycle stretch and hypoxia) conditions and exposed to NiONPs (0.5–5 μg/cm2) for 4 or 24 h. The following endpoints were studied: (i) ROS production using CM-H2DCF-DA and MitoSOX probes, (ii) nitrite production by the Griess reaction, (iii) IL-6 secretion by ELISA, (iv) calcium signaling with a Fluo-4 AM probe, and (v) mitochondrial dysfunction with TMRM and MitoTracker probes. Our results evidenced that under pathological conditions, ROS and nitrite production, IL-6 secretions, calcium signaling, and mitochondria alterations increased compared to physiological conditions. Human exposure to NiONPs may be associated with adverse effects in vulnerable populations with cardiovascular risks.

The pulmonary effects of nickel-containing nanoparticles: cytotoxicity, genotoxicity, carcinogenicity, and their underlying mechanisms

“…37,43 Increased ROS generation was observed in BEAS-2B, but not A549, cells exposed to 60 and 100 μg mL −1 NiO NPs for 45 min 31 or 25 and 50 μg Ni mL −1 of NiO NPs for 5 min to 2 h. 19,24 Others found increased ROS, perturbation of the mitochondrial membrane potential, lipid peroxidation, increased tHODE levels, or depletion of antioxidants, such as decreased GSH, SOD, or CAT, in A549 cells exposed to NiO NPs. 30,35,44 NiO NP exposure also caused ROS and nitrite production and mitochondrial impairment in human pulmonary artery endothelial cells HPAEC, 41,45 as well as upregulation of HO-1 in A549 44,46 and 16HBE14o cells. 46 NiO NP exposure also caused cell apoptosis, necrosis, or suppression of cell proliferation in A549, 30,31 H460, 20 HEp-2, 38 BEAS-2B, 24,31,40 HPAEpiC, 40 and HPAEC cells.…”

“…41 Di Bucchianico et al reported that NiO NP-induced BEAS-2B cell apoptosis and necrosis were calcium-dependent, 24 but Duan et al reported that they were via p53 hyperacetylation and Bax activation by downregulation of SIRT1. 39 NiO NP exposure also caused increased cytosolic calcium (Ca 2+ ) concentration in human pulmonary artery endothelial cells HPAEC, 41,45 which was responsible for NiO NP-induced cell apoptosis. Liu et al demonstrated that NiO NP exposure induced apoptosis and ferroptosis in BEAS-2B and HPAEpiC cells via ATF3.…”

“…33 NiO NP exposure also caused IL-6 production in human pulmonary artery endothelial cells HPAEC. 41,45 Yang et al reported that NiO NP-induced inflammation was via downregulation of lncRNA MEG3 and activation of the p38 MAPK pathway in A549 cells. 47 NiO NPs were able to cause human lung cells to undergo epithelial-mesenchymal transition (EMT) and upregulation of fibrosis-associated proteins.…”

HYDROGEN PREVENTS LIPOPOLYSACCHARIDE-INDUCED PULMONARY MICROVASCULAR ENDOTHELIAL CELL INJURY BY INHIBITING STORE-OPERATED Ca2+ ENTRY REGULATED BY STIM1/ORAI1

“…Variations in intracellular Ca 2+ were detected using Fluo-4 AM green dye (Molecular Probes; Invitrogen Life Technologies, Carlsbad, CA) according to the manufacturer’s protocol (33). Pulmonary microvascular endothelial cells were subjected to different treatments according to the experimental procedures.…”