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. 2023 Mar 6;14:1114739. doi: 10.3389/fphar.2023.1114739

FIGURE 6.

FIGURE 6

Aerosolized PHMG induced barrier disruption and cytotoxicity in triple-cell culture on-chip (A) Timeline and schematic of PHMG nebulization at day 21 in tri-cell culture (AXiAECs/d-THP1/HLMVEC) model on-chip. (B) Representative immunofluorescence staining for apical alveolar and basal endothelial barrier after 24 h of PHMG exposure. Cells were analyzed for tight junction, ZO1 (green) and nuclei with Hoechst (blue). Scale bar is 50 µm. (C) TER values (Ohm-cm2) were recorded before PHMG nebulization (at 0 h) and at 4 h, 24 h and 48 h after exposure (N = 3; n = 12). (D) Cytotoxicity was calculated from LDH release at 24 h and 48 h after PHMG nebulization from both ALI and ALI + CS conditions. LDH release was normalized with respective untreated controls. Data shown as mean ± SEM (N = 1; n = 3/condition/timepoint). (E) Gene expression after 48 h of exposure was assessed for cells in ALI + CS condition. Fold change values were normalized with respective untreated controls. Data presented as mean ± SEM (N = 3; n = 6).