Development of In Vitro Co-Culture Model to Mimic the Cell to Cell Communication in Response to Urban PM_2.5

Abstract

Background Airborne particulate matter (PM), a widespread air contaminant, is a complex mixture of solids and aerosols composed of particles suspended in the air. PM is associated with inflammatory responses and may worsen inflammatory skin diseases. However, the mechanisms through which PM affects atopic dermatitis (AD) remain unclear. Objective To establish an in vitro model that more accurately mimics AD using human keratinocyte (HaCaT), dermal fibroblast (HDF), and mast cell (HMC-1) and using this model to investigate the mechanism through which PMs affect AD. Methods An AD-like in vitro model was established by seeding HaCaT, HDF, and HMC-1 cells with recombinant human interleukin (IL)-1α and polyinosinic:polycytidylic acid. We confirmed the effect of PM on the inflammatory cytokine expression of a triple-cell culture model. SRM 1649b Urban Dust, which is mainly composed of polycyclic aromatic hydrocarbons, was used as the reference PM. The effects of PM on the expression levels of proinflammatory cytokines and skin barrier markers were assessed using quantitative real-time polymerase chain reaction and western blotting. Inflammatory cytokine levels were measured using an enzyme-linked immunosorbent assay. Results Interactions between various skin cell types were evaluated using a co-culture system. PM treatment increased mRNA and protein levels of the inflammatory cytokines IL-6, IL-1α, tumor necrosis factor-α, IL-4, and IL-1β and decreased the expression of the skin barrier markers filaggrin and loricrin. Conclusion Our results suggest that an in vitro triple-cell culture model using HaCaT, HDF, and HMC-1 cells may be reliable for obtaining more physiological, functional, and reproducible data on AD and skin barriers.

Epigenetic mechanisms of particulate matter exposure: air pollution and hazards on human health

“…These findings suggest that PM 2.5 exposure is a risk factor for lung cancer progression and that the AhR-TMPRSS2-IL18 pathway is a potential target for the development of new therapies against this disease ( Wang et al, 2023 ). Moreover, in an innovative in vitro model that simulates cell interactions occurring in skin during atopic dermatitis, it was shown that PM 2.5 exposure led to an upregulation of proinflammatory cytokines, including IL-1α, IL-1β, IL-6, IL-4, IL-13, and TNF-α in a co-culture assay ( Roh et al, 2022 ).…”

Adipose tissue-derived exosomes alleviate particulate matter-induced inflammatory response and skin barrier damage in atopic dermatitis-like triple-cell model

“…The triple-cell culture was pretreated with Poly I:C (10 μg/mL) and IL-1α (10 ng/mL) to induce an AD-like phenotype. After 24 hours, the cell cultures were treated with ASC-exosomes and PM at a concentration of 25 μg/cm 2 , according to previous studies [7,8,12]. The plates were incubated at 37˚C for an additional 24 hours.…”

“…An AD-like triple-cell model with three different cell types, including HaCaT, HDF, and HMC-1 cells, was used to investigate the cell-to-cell interactions associated with AD [12]. We compared the intracellular uptake of PKH67-labeled ASC-exosomes in HaCaT cells (Fig 1A Therefore, the treatment time of the exosomes was selected as 24 and 48 hours.…”

“…The contribution of PM exposure to health problems [8,9], such as cardiovascular, nervous, immune, and respiratory diseases, is increasingly becoming an area of interest [5,9]. For example, the effects of PM (e.g., fine dust) on skin diseases have been studied more often in recent years [10][11][12]. Our laboratory's previous research has shown that PM is involved in skin barrier defects and abnormal immune responses [12][13][14][15][16], and we advocated for the development of therapeutic agents for skin barrier repair and skin inflammation reduction in response to PM exposure [12,15,17].…”

“…However, the effects of ASC-derived exosomes on AD remain poorly understood. In this study, the effectiveness and mechanisms of ASC-derived exosomes on AD exacerbation by air pollutant PM were investigated using an AD-like cell model [12,[29][30][31].…”