A Novel Nanosafety Approach Using Cell Painting, Metabolomics, and Lipidomics Captures the Cellular and Molecular Phenotypes Induced by the Unintentionally Formed Metal-Based (Nano)Particles

Abstract

Additive manufacturing (AM) or industrial 3D printing uses cutting-edge technologies and materials to produce a variety of complex products. However, the effects of the unintentionally emitted AM (nano)particles (AMPs) on human cells following inhalation, require further investigations. The physicochemical characterization of the AMPs, extracted from the filter of a Laser Powder Bed Fusion (L-PBF) 3D printer of iron-based materials, disclosed their complexity, in terms of size, shape, and chemistry. Cell Painting, a high-content screening (HCS) assay, was used to detect the subtle morphological changes elicited by the AMPs at the single cell resolution. The profiling of the cell morphological phenotypes, disclosed prominent concentration-dependent effects on the cytoskeleton, mitochondria, and the membranous structures of the cell. Furthermore, lipidomics confirmed that the AMPs induced the extensive membrane remodeling in the lung epithelial and macrophage co-culture cell model. To further elucidate the biological mechanisms of action, the targeted metabolomics unveiled several inflammation-related metabolites regulating the cell response to the AMP exposure. Overall, the AMP exposure led to the internalization, oxidative stress, cytoskeleton disruption, mitochondrial activation, membrane remodeling, and metabolic reprogramming of the lung epithelial cells and macrophages. We propose the approach of integrating Cell Painting with metabolomics and lipidomics, as an advanced nanosafety methodology, increasing the ability to capture the cellular and molecular phenotypes and the relevant biological mechanisms to the (nano)particle exposure.

NLRP3 inflammasome as a sensor of micro- and nanoplastics immunotoxicity

“…It is also important to emphasize the need, in experimental studies, to “coat” MNPs with the relevant biomolecular corona (MNPs never interact with cells as pristine particles), to use realistic MNP concentrations, and to perform longitudinal studies (to assess whether inflammasome activation becomes persistent or not). In addition, novel approaches in the nanosafety field, such as high-content screening combined with multi-omics ( 108 ), may be helpful tools in dissecting the cellular and molecular phenotypes upon inflammasome activation by MNPs.…”

Nanoparticles‐induced potential toxicity on human health: Applications, toxicity mechanisms, and evaluation models

“…Quantitative structure‐property connections are used to characterize a variety of theoretical modeling approaches used to predict the physiochemical properties of nano molecules 397 . Multiomics 398 approaches including genomics, transcriptomics, proteomics, and metabolomics are possible to anticipate new biological pathways that are related to nanotoxicity and to identify cytotoxicity at low exposure levels to NPs 339,399 …”

“…397 Multiomics 398 approaches including genomics, transcriptomics, proteomics, and metabolomics are possible to anticipate new biological pathways that are related to nanotoxicity and to identify cytotoxicity at low exposure levels to NPs. 339,399 There are still some limitations and challenges with the current models used to assess the toxicity of nanomaterials. For example, organ-on-a-chip technology and 3D cell culture models are more complex and expensive than traditional toxicity testing methods, which may limit their widespread use.…”

Health hazards of particles in additive manufacturing: a cross-disciplinary study on reactivity, toxicity and occupational exposure to two nickel-based alloys

“…In another recent study, we focused on spatter/condensate particles and showed also low cytotoxic, genotoxic, and inflammatory potential of these particles 18 . On the other hand, subtle morphological changes, such as concentration-dependent effects on the cytoskeleton were observed in a recent study on iron-based AM particles 19 . More studies are needed regarding inhalable particles (< 4 µm) that more easily can reach deep into the lung.…”