2D MATERIALS INTERACTION WITH IMMUNE CELLS: A PILOT STUDY ON BISMUTHENE

Bismuthene is a potential nanomaterial for upcoming biomedical applications such as biosensing and drug delivery due to its exceptional electrical, topological, thermoelectric, and optical properties. Assessing the nanomaterial’s immunocompatibility is a fundamental step for its translation into the clinic. We investigated the interactions of bismuthene with peripheral blood mononuclear cells (PBMCs) and whole human blood through a wide variety of assays, including high dimensional analysis by single-cell mass cytometry. First, we performed a hemolysis assay on whole blood, and blood cell counts. Then viability, necrosis, and apoptosis assays were performed on PBMCs bismuthene treated on dose and time response. The immune cell activation is a key parameter of the immune response functionality. We measured the potential cell activation after material treatment by testing the production and release of a broad range of cytokines such as IL-4, IL-5, IL-6, IL-10, IL-12, TNFα, and INFγ by flow cytometry arrays. In addition, the functionality of T cells and monocytes, representative populations of the adaptive and innate immune response, were evaluated by looking at the expression of two essential activation markers: CD25 and CD69. To deepen the understanding of the potential immune interactions with a large variety of cell populations, we adapted a protocol for high-dimensional nano-immune profiling for the first time in the context of nanotechnology. We dissected the bismuthene biocompatibility of 37 different immune subpopulations by single-cell mass cytometry. Our results suggest that bismuthene is a highly immune-compatible material, it does not lead to blood hemolysis, cell immune activation, or cytotoxicity in any of the analyzed cell types. Moreover, we established a new technological pipeline in the context of immuno-nanotoxicology, which can be adopted to assess the immune compatibility of several other nanomaterials by simultaneously dissecting up to 37 immune cell types.