Comprehensive analysis of resilience of human airway epithelial barrier against short-term PM2.5 inorganic dust exposure using invitro microfluidic chip and exvivo human airway models.

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The updated World Health Organization (WHO) air quality guideline recommends an annual mean concentration of fine particulate matter (PM2.5) not exceeding 5 or 15 μg/m3 in the short-term (24 h) for no more than 3-4 days annually. However, more than 90% of the global population is currently exposed to daily concentrations surpassing these limits, especially during extreme weather conditions and due to transboundary dust transport influenced by climate change. Herein, the effect of respirable <PM2.5 inorganic silica particle exposures on epithelial barrier integrity was simultaneously evaluated within the biomimetic microfluidic platform-based airway epithelial barrier (AEB)-on-a-chip and human bronchoscopic exvivo airway tissue models, comparatively. Silica particles at an average size of 1 μm, referred to as <PM2.5, dose-dependently tested by MTT and LDH analyses. The elicited dose of 800 μg/mL was applied to human airway epithelial cells (Calu-3) seeded to the membrane at air-liquid interface in the AEB-on-a-chip platform, which is operated under static and dynamic conditions and to exvivo human bronchoscopy bronchial tissue slices for 72 h. For both models, healthy and exposed groups were comparatively investigated. Computational fluid dynamics simulations were performed to assess shear stress profiles under different flow conditions. Qualitative and quantitative analyses were carried out to evaluate the resilience of the epithelial barrier via cell survivability, morphology, barrier integrity, permeability, and inflammation. In the AEB-on-a-chip platform, short-term exposure to 800 μg/mL PM2.5 disrupted AEB integrity via increasing barrier permeability, decreasing cell adhesion-barrier markers such as ZO-1, Vinculin, ACE2, and CD31, impaired cell viability and increased the expression levels of proinflammatory markers; IFNs, IL-6, IL-1s, TNF-α, CD68, CD80, and Inos, mostly under dynamic conditions. Besides, decreased tissue viability, impaired tissue integrity via decreasing of Vinculin, ACE2, β-catenin, and E-cadherin, and also proinflammatory response with elevated CD68, IL-1α, IL-6, IFN-Ɣ, Inos, and CD80 markers, were observed after PM2.5 exposure in exvivo tissue. The duration and concentration of PM2.5 that can be exposed during extreme weather conditions and natural events aligns with our exposure model (0-800 μg/mL 72 h). At this level of exposure, the resilience of the epithelial barrier is demonstrated by both AEB-on-a-chip platform emulating dynamic forces in the body and exvivo bronchial biopsy slices. Lung-on-a-chip models will serve as reliable exposure models in this context.