Donor-to-donor variability of a human three-dimensional bronchial epithelial model: A case study of cigarette smoke exposure

Mori, Sakura; Ishimori, Kanae; Matsumura, Kazunori; Ishikawa, Shinkichi; Ito, Shigeaki

doi:10.1016/j.tiv.2022.105391

Cited by 10 publications

(7 citation statements)

References 39 publications

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“…Within each timepoint, all cartilage explants were generated from the same animal to reduce differences due to animal‐to‐animal variability within a timepoint (i.e., all day 0 samples were from one animal and all day 21 samples from another animal). This donor to donor variability has been reported in cell isolation studies as well as tissue grafting 37–39 . While this experimental design choice posed a constraint on the number of samples per experimental group, this permitted more direct interrogation of how material charge and LCST influence tissue integration and cellular infiltration.…”

Section: Discussionmentioning

confidence: 95%

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Thermogelling hydrogel charge and lower critical solution temperature influence cellular infiltration and tissue integration in an ex vivo cartilage explant model

Pearce

Swain

Victor

et al. 2022

J Biomedical Materials Res

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Thermogelling hydrogels based on poly(N-isopropyl acrylamide) (p [NiPAAm]) and crosslinked with a peptide-bearing macromer poly(glycolic acid)-poly(ethylene glycol)-poly(glycolic acid)-di(but-2-yne-1,4-dithiol) (PdBT) were fabricated to assess the role of hydrogel charge and lower critical solution temperature (LCST) over time in influencing cellular infiltration and tissue integration in an ex vivo cartilage explant model over 21 days. The p(NiPAAm)-based thermogelling polymer was synthesized to possess 0, 5, and 10 mol% dimethyl-γ-butyrolactone acrylate (DBA) to raise the LCST over time as the lactone rings hydrolyzed. Further, three peptides were designed to impart charge into the hydrogels via conjugation to the PdBT crosslinker.The positively, neutrally, and negatively charged peptides K4 (+), zwitterionic K2E2 (0), and E4 (À), respectively, were conjugated to the modular PdBT crosslinker and the hydrogels were evaluated for their thermogelation behavior in vitro before injection into the cartilage explant models. Samples were collected at days 0 and 21, and tissue integration and cellular infiltration were assessed via mechanical pushout testing and histology. Negatively charged hydrogels whose LCST changed over time (10 mol% DBA) were demonstrated to promote the greatest tissue integration when compared to the positive and neutral gels of the same thermogelling polymer formulation due to increased transport and diffusion across the hydrogel-tissue interface.Indeed, the negatively charged thermogelling polymer groups containing 5 and 10 mol% DBA demonstrated cellular infiltration and cartilage-like matrix deposition via histology. This study demonstrates the important role that material physicochemical properties play in dictating cell and tissue behavior and can inform future cartilage tissue engineering strategies.

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Section: Discussionmentioning

confidence: 95%

“…This donor to donor variability has been reported in cell isolation studies as well as tissue grafting. [37][38][39] While this experimental design choice posed a constraint on the number of samples per experimental group, this…”

Section: Discussionmentioning

confidence: 99%

Thermogelling hydrogel charge and lower critical solution temperature influence cellular infiltration and tissue integration in an ex vivo cartilage explant model

Pearce

Swain

Victor

et al. 2022

J Biomedical Materials Res

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“…These results suggest donor‐dependent differences in the threshold of EGFR activation via ligand binding, time‐course of ligand binding to receptor activation, or ligand‐independent phosphorylation (Filosto et al, 2012). The variations observed in primary cell cultures might be explained by the original characteristics of the donors (Bovard et al, 2020; Mori et al, 2022; Rayner et al, 2019). Theoretically, not all individuals have the same transcriptomic, proteomic, and epigenetic characteristics (Jackson et al, 2020; Stefanowicz et al, 2012); therefore, the capacity of the antioxidant system might also vary.…”

Section: Discussionmentioning

confidence: 99%

Oxidative stress‐mediated epidermal growth factor receptor activation by cigarette smoke or heated tobacco aerosol in human primary bronchial epithelial cells from multiple donors

Muratani

Ichikawa

Erami

et al. 2023

J of Applied Toxicology

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The epidermal growth factor receptor (EGFR) signaling pathway has essential roles in maintaining homeostasis of various tissues by regulating cell proliferation and differentiation. Deregulation of the EGFR signaling pathway is associated with various chronic diseases including chronic obstructive pulmonary disease. Cigarette smoke (CS) is known to activate EGFR, which is linked to chronic obstructive pulmonary disease. The biological sequence from CS exposure to EGFR activation is initiated by oxidative stress caused by intracellular reactive oxygen species (ROS) and the depletion of glutathione, which led to EGFR ligand secretion and EGFR activation.We hypothesized that reducing exposure to CS constituents contributes to preventing CS-inducible EGFR activation. Therefore, we examined the aerosol from heated tobacco products (HTPs) because the aerosol contains fewer chemical constituents at lower levels than CS. We exposed primary human bronchial epithelial cells from four donors to the extracts of CS from a 1R6F reference cigarette or HTP aerosol from three in-market products, including our DT3.0a. The biological sequence from ROS to EGFR activation was assessed. CS induced all the tested endpoints although inter-donor differences were observed, whereas HTPs elicited most of the biological events at higher concentrations; however, EGFR phosphorylation was not observed even at fivefold higher concentration than CS. Overall, our results indicate that HTPs are less effective than CS to elicit ROS-induced EGFR activation. The reduced-risk potential of HTPs on EGFR-related diseases should be investigated further. In addition, testing with multiple donors is warranted when considering the individual differences in responses of primary cells to stimuli.

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“…It has been reported that RhBE secretes IL-8 at ng/ml levels in basolateral media. 48,49 Thus, most of the IL-8 detected in the cocultures is likely to be derived from the RhBE. The recently proposed AOP for respiratory sensitisation indicates that cellular damage to the epithelium and the secretion of proinflammatory mediators from the epithelium could be key events during the induction of respiratory sensitisation.…”

Section: Discussionmentioning

confidence: 99%

Development of an In Vitro Sensitisation Test Using a Coculture System of Human Bronchial Epithelium and Immune Cells

Tanabe,

Yoshida,

Ishikawa

et al. 2023

Altern Lab Anim

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Chemical respiratory sensitisation is a serious health problem. However, to date, there are no validated test methods available for identifying respiratory sensitisers. The aim of this study was to develop an in vitro sensitisation test by modifying the human cell line activation test (h-CLAT) to detect respiratory sensitisers and distinguish them from skin sensitisers. THP-1 cells were exposed to the test chemicals (two skin sensitisers and six respiratory sensitisers), either as monocultures or as cocultures with air–liquid interface-cultured reconstructed human bronchial epithelium. The responses were analysed by measuring the expression levels of surface markers on THP-1 cells (CD86, CD54 and OX40L) and the concentrations of cytokines in the culture media (interleukin (IL)-8, IL-33 and thymic stromal lymphopoietin (TSLP)). The cocultures exhibited increased CD54 expression on THP-1 cells; moreover, in the cocultures but not in the monocultures, exposure to two uronium salts (i.e. respiratory sensitisers) increased CD54 expression on THP-1 cells to levels above the criteria for a positive h-CLAT result. Additionally, exposure to the respiratory sensitiser abietic acid, significantly increased IL-8 concentration in the culture medium, but only in the cocultures. Although further optimisation of the method is needed to distinguish respiratory from skin sensitisers by using these potential markers (OX40L, IL-33 and TSLP), the coculture of THP-1 cells with bronchial epithelial cells offers a potentially useful approach for the detection of respiratory sensitisers.

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Donor-to-donor variability of a human three-dimensional bronchial epithelial model: A case study of cigarette smoke exposure

Cited by 10 publications

References 39 publications

Thermogelling hydrogel charge and lower critical solution temperature influence cellular infiltration and tissue integration in an ex vivo cartilage explant model

Thermogelling hydrogel charge and lower critical solution temperature influence cellular infiltration and tissue integration in an ex vivo cartilage explant model

Oxidative stress‐mediated epidermal growth factor receptor activation by cigarette smoke or heated tobacco aerosol in human primary bronchial epithelial cells from multiple donors

Development of an In Vitro Sensitisation Test Using a Coculture System of Human Bronchial Epithelium and Immune Cells

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