Journal: Scientific Reports

Article Title: A next-generation system for smoke inhalation integrated with a breathing lung-on-chip to model human lung responses to cigarette exposure

doi: 10.1038/s41598-025-00438-z

Figure Lengend Snippet: Apical surfactant addition demonstrates potential protective effects against CS exposure. ( A ) Overview of cells used in the triple culture “TC THP1” (AXiAECs/hLMVECs/THP1), culture conditions (dynamic; ALI + Str) and treatment (Surfactant pre-treated followed by CS exposure) performed. ( B ) Representative immunofluorescence staining of CTRL ALI + Str (− Surfactant) cells (top row; scale bar 50 µm), CS exposed ALI + Str (-Surfactant) cells (second row; scale bar 50 µm), CTRL ALI + Str (+ Surfactant) cells (third row; scale bar 100 µm), CS exposed ALI + Str (+ Surfactant) cells (fourth row; scale bar 100 µm). Cells were stained for Zonula Occludens 1 (ZO-1, in green) and nuclei (Hoechst, in blue). ( C ) IL-8 concentration (pg/ml) measured from apical supernatants collected from the CTRL and CS exposed wells pre-treated with and without surfactant (+ Surf or -Surf) via ELISA assay (N = 2; n = 6/condition). ( D ) mRNA was harvested at 48 h time-point with and without surfactant (+ Surf or -Surf) post-exposure to CS. qRT-PCR study for inflammation (Interleukin 6, IL6; Interleukin 8, IL8; Tumor Necrosis Factor α, TNFα) and oxidative stress-related (Heme Oxygenase 1, HO-1; Human MutT Homolog 1, MTH-1) genes were conducted in all -Surf and + Surf CS exposed and CTRL samples (N = 2, n = 3). Data are shown as mean ± SEM.

Article Snippet: The AX iAECs were cultured according to the manufacturer’s instructions, as previously described and characterized by Sengupta et al. . Human Lung Microvascular Endothelial Cells (hLMVECs) purchased from PromoCell were expanded in flasks using AX Endothelial Medium. hLMVECs were first seeded on the basolateral side of each membrane in the AX12 at a density of 106,000cells/cm 2 , followed by a 2-h incubation, and then AX iAECs were seeded apically at a density of 409,000 cells/cm 2 on day 0 for all co-culture experiments.

Techniques: Immunofluorescence, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: A next-generation system for smoke inhalation integrated with a breathing lung-on-chip to model human lung responses to cigarette exposure

doi: 10.1038/s41598-025-00438-z

Figure Lengend Snippet: Differential effects of CS exposure on various co-culture dynamic models. ( A ) Overview of cells used in the dual culture “DC” ( AX iAECs/hLMVECs), culture conditions (dynamic; ALI + Str) and treatment (CS exposure) performed. ( B ) Overview of cells used in the triple culture “TC pBDMs” ( AX iAECs/hLMVECs/pBDMs) and “TC THP1” ( AX iAECs/hLMVECs/THP1), culture conditions (dynamic; ALI + Str) and treatment (CS exposure) performed. ( C ) TER (Ohm cm 2 ) measured in “ALI + Str” conditions pre-exposure at 0 h and 4 h, 24 h and 48 h post-exposure to CS in DC (N = 3; n = 3/time-point), in TC pBDMs (N = 2; n = 3/time-point) and TC THP1 (N = 3; n = 3/time-point). Data are shown as violin plots, medians are indicated by black dotted lines. ( D ) The levels of IL-8 (pg/ml) in the supernatants collected from the cells were measured by ELISA in all the co-culture models. ( E ) Cytotoxicity was calculated from LDH release at 48 h time-point after CS exposure (N = 2; n = 3/time-point for all models). Data are shown as mean ± SEM.

Article Snippet: The AX iAECs were cultured according to the manufacturer’s instructions, as previously described and characterized by Sengupta et al. . Human Lung Microvascular Endothelial Cells (hLMVECs) purchased from PromoCell were expanded in flasks using AX Endothelial Medium. hLMVECs were first seeded on the basolateral side of each membrane in the AX12 at a density of 106,000cells/cm 2 , followed by a 2-h incubation, and then AX iAECs were seeded apically at a density of 409,000 cells/cm 2 on day 0 for all co-culture experiments.

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay