Boron Nitride Nanosheets Induce Lipid Accumulation and Autophagy in Human Alveolar Lung Epithelial Cells Cultivated at Air‐Liquid Interface

Hexagonal boron nitride (hBN) is an emerging 2D material attracting significant attention due to its superior electrical, chemical, and therapeutic properties. However, inhalation toxicity mechanisms of hBN in human lung cells are poorly understood. Here, cellular interaction and effects of hBN nanosheets is investigated in alveolar epithelial cells cultured on porous inserts and exposed under air−liquid interface conditions for 24 h. hBN is taken up by the cells as determined in a label‐free manner via RAMAN‐confocal microscopy, ICP‐MS, TEM, and SEM‐EDX. No significant (p > 0.05) effects are observed on cell membrane integrity (LDH release), epithelial barrier integrity (TEER), interleukin‐8 cytokine production or reactive oxygen production at tested dose ranges (1, 5, and 10 µg cm−2). However, it is observed that an enhanced accumulation of lipid granules in cells indicating the effect of hBN on lipid metabolism. In addition, it is observed that a significant (p < 0.05) and dose‐dependent (5 and 10 µg cm−2) induction of autophagy in cells after exposure to hBN, potentially associated with the downstream processing and breakdown of excess lipid granules to maintain lipid homeostasis. Indeed, lysosomal co‐localization of lipid granules supporting this argument is observed. Overall, the results suggest that the continuous presence of excess intracellular lipids may provoke adverse outcomes in the lungs. The present study demonstrates the uptake of hexagonal boron nitride (hBN) nanosheets by alveolar lung cells, which subsequently triggers subtoxic effects via intracellular accumulation of lipid granules followed by autophagy induction that potentially protects the cells from acute lipotoxicity.