Single-cell mass cytometry reveals cell type- and cluster-specific heterogeneity in silver nanoparticle responses in a 3D alveolar tetra-culture model

Utilizing a combination of a physiologically relevant 3D alveolar co-culture model and single-cell mass cytometry (CyTOF), this research seeks to elucidate the heterogeneous cellular responses to silver nanoparticles (AgNPs) across multiple human lung-associated cell types. By employing hierarchical clustering, we aim to uncover cluster-specific heterogeneity and explore how distinct cell subpopulations engage in both common and unique responses, ultimately revealing the intracellular signaling dynamics that determine survival or apoptosis following nanoparticle exposure. This study integrated a 3D alveolar model with single-cell analysis to reveal the heterogeneous cellular responses to silver nanoparticles (AgNPs). The findings show that cytotoxicity is governed more by complex intracellular signaling—such as inflammation and stress adaptation—than by nanoparticle association alone, with THP-1 cells exhibiting unique inflammation-driven apoptotic pathways. These results underscore the importance of accounting for cellular heterogeneity and intercellular communication when assessing nanoparticle-induced toxicity.