Functional characterisation and application of an ex vivo perfusion-superfusion system in murine airways

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Abstract

Introduction The aim of this study was to develop two dynamic ex vivo airway explant systems, a perfusion-superfusion system and a ventilation-superfusion system, for the study of toxic airborne substances, such as the prevalent smoke constituent acrolein. Methods Mouse isolated tracheal segments were perfused with physiological media or ventilated with humidified air at 37 °C to mimic dynamic flow conditions, and superfused with media over the exterior surface. At selected time points, the histological and functional integrity of segments was evaluated. The perfusion-superfusion system was subsequently used to examine mucin secretory responses elicited by acrolein in airways in which mucous metaplasia had been induced with lipopolysaccharide (LPS; 1 μg ml− 1) prior to 24 h of media perfusion, followed by stimulation with acrolein or ATP for 15 min. Epithelial mucin levels were determined by quantitative analysis of periodic acid-Schiff's reagent (PAS)-stained sections. Results Epithelial morphology was successfully preserved in the perfusion-superfusion and ventilation-superfusion systems for at least 24 h and up to 18 h, respectively. At these time points, the contractile and relaxation responses of perfused and ventilated tracheal segments to carbachol, the neuropeptide substance P, and the prostanoid PGE2 were also preserved. Using the perfusion-superfusion system, acute exposure to acrolein caused a dose-dependent reduction in the levels of PAS-positive mucin stores induced by LPS, consistent with mucin secretion. Discussion Both the perfusion-superfusion and ventilation-superfusion systems successfully preserved the viability of mouse isolated tracheal segments on a histological and functional level, and the perfusion-superfusion system was used to characterise the mucin secretory responses elicited by acrolein. Thus, this system may be a useful model through which to conduct further toxicological studies in mammalian airways.

Original languageEnglish
Pages (from-to)66-77
Number of pages12
JournalJournal of Pharmacological and Toxicological Methods
Volume84
DOIs
Publication statusPublished - 1 Mar 2017

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Acrolein
Mucins
Perfusion
Ventilation
Periodic Acid
Poisons
Carbachol
Substance P
Neuropeptides
Dinoprostone
Smoke
Prostaglandins
Lipopolysaccharides
Metaplasia
Adenosine Triphosphate
Toxicology
Air
Chemical analysis

Cite this

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title = "Functional characterisation and application of an ex vivo perfusion-superfusion system in murine airways",
abstract = "Introduction The aim of this study was to develop two dynamic ex vivo airway explant systems, a perfusion-superfusion system and a ventilation-superfusion system, for the study of toxic airborne substances, such as the prevalent smoke constituent acrolein. Methods Mouse isolated tracheal segments were perfused with physiological media or ventilated with humidified air at 37 °C to mimic dynamic flow conditions, and superfused with media over the exterior surface. At selected time points, the histological and functional integrity of segments was evaluated. The perfusion-superfusion system was subsequently used to examine mucin secretory responses elicited by acrolein in airways in which mucous metaplasia had been induced with lipopolysaccharide (LPS; 1 μg ml− 1) prior to 24 h of media perfusion, followed by stimulation with acrolein or ATP for 15 min. Epithelial mucin levels were determined by quantitative analysis of periodic acid-Schiff's reagent (PAS)-stained sections. Results Epithelial morphology was successfully preserved in the perfusion-superfusion and ventilation-superfusion systems for at least 24 h and up to 18 h, respectively. At these time points, the contractile and relaxation responses of perfused and ventilated tracheal segments to carbachol, the neuropeptide substance P, and the prostanoid PGE2 were also preserved. Using the perfusion-superfusion system, acute exposure to acrolein caused a dose-dependent reduction in the levels of PAS-positive mucin stores induced by LPS, consistent with mucin secretion. Discussion Both the perfusion-superfusion and ventilation-superfusion systems successfully preserved the viability of mouse isolated tracheal segments on a histological and functional level, and the perfusion-superfusion system was used to characterise the mucin secretory responses elicited by acrolein. Thus, this system may be a useful model through which to conduct further toxicological studies in mammalian airways.",
keywords = "Acrolein, ATP, Ex vivo system, Methods, Models, Mouse trachea, Mucin secretion, Perfusion, Superfusion, Ventilation",
author = "Cheah, {Esther Y.} and Mann, {Tracy S.} and Burcham, {Philip C.} and Henry, {Peter J.}",
year = "2017",
month = "3",
day = "1",
doi = "10.1016/j.vascn.2016.11.003",
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volume = "84",
pages = "66--77",
journal = "Journal of Pharmacological and Toxicological Methods",
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TY - JOUR

T1 - Functional characterisation and application of an ex vivo perfusion-superfusion system in murine airways

AU - Cheah, Esther Y.

AU - Mann, Tracy S.

AU - Burcham, Philip C.

AU - Henry, Peter J.

PY - 2017/3/1

Y1 - 2017/3/1

N2 - Introduction The aim of this study was to develop two dynamic ex vivo airway explant systems, a perfusion-superfusion system and a ventilation-superfusion system, for the study of toxic airborne substances, such as the prevalent smoke constituent acrolein. Methods Mouse isolated tracheal segments were perfused with physiological media or ventilated with humidified air at 37 °C to mimic dynamic flow conditions, and superfused with media over the exterior surface. At selected time points, the histological and functional integrity of segments was evaluated. The perfusion-superfusion system was subsequently used to examine mucin secretory responses elicited by acrolein in airways in which mucous metaplasia had been induced with lipopolysaccharide (LPS; 1 μg ml− 1) prior to 24 h of media perfusion, followed by stimulation with acrolein or ATP for 15 min. Epithelial mucin levels were determined by quantitative analysis of periodic acid-Schiff's reagent (PAS)-stained sections. Results Epithelial morphology was successfully preserved in the perfusion-superfusion and ventilation-superfusion systems for at least 24 h and up to 18 h, respectively. At these time points, the contractile and relaxation responses of perfused and ventilated tracheal segments to carbachol, the neuropeptide substance P, and the prostanoid PGE2 were also preserved. Using the perfusion-superfusion system, acute exposure to acrolein caused a dose-dependent reduction in the levels of PAS-positive mucin stores induced by LPS, consistent with mucin secretion. Discussion Both the perfusion-superfusion and ventilation-superfusion systems successfully preserved the viability of mouse isolated tracheal segments on a histological and functional level, and the perfusion-superfusion system was used to characterise the mucin secretory responses elicited by acrolein. Thus, this system may be a useful model through which to conduct further toxicological studies in mammalian airways.

AB - Introduction The aim of this study was to develop two dynamic ex vivo airway explant systems, a perfusion-superfusion system and a ventilation-superfusion system, for the study of toxic airborne substances, such as the prevalent smoke constituent acrolein. Methods Mouse isolated tracheal segments were perfused with physiological media or ventilated with humidified air at 37 °C to mimic dynamic flow conditions, and superfused with media over the exterior surface. At selected time points, the histological and functional integrity of segments was evaluated. The perfusion-superfusion system was subsequently used to examine mucin secretory responses elicited by acrolein in airways in which mucous metaplasia had been induced with lipopolysaccharide (LPS; 1 μg ml− 1) prior to 24 h of media perfusion, followed by stimulation with acrolein or ATP for 15 min. Epithelial mucin levels were determined by quantitative analysis of periodic acid-Schiff's reagent (PAS)-stained sections. Results Epithelial morphology was successfully preserved in the perfusion-superfusion and ventilation-superfusion systems for at least 24 h and up to 18 h, respectively. At these time points, the contractile and relaxation responses of perfused and ventilated tracheal segments to carbachol, the neuropeptide substance P, and the prostanoid PGE2 were also preserved. Using the perfusion-superfusion system, acute exposure to acrolein caused a dose-dependent reduction in the levels of PAS-positive mucin stores induced by LPS, consistent with mucin secretion. Discussion Both the perfusion-superfusion and ventilation-superfusion systems successfully preserved the viability of mouse isolated tracheal segments on a histological and functional level, and the perfusion-superfusion system was used to characterise the mucin secretory responses elicited by acrolein. Thus, this system may be a useful model through which to conduct further toxicological studies in mammalian airways.

KW - Acrolein

KW - ATP

KW - Ex vivo system

KW - Methods

KW - Models

KW - Mouse trachea

KW - Mucin secretion

KW - Perfusion

KW - Superfusion

KW - Ventilation

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U2 - 10.1016/j.vascn.2016.11.003

DO - 10.1016/j.vascn.2016.11.003

M3 - Article

VL - 84

SP - 66

EP - 77

JO - Journal of Pharmacological and Toxicological Methods

JF - Journal of Pharmacological and Toxicological Methods

SN - 1056-8719

ER -