Protocol for constructing an accessible exposure chamber for in vitro and in vivo modeling of airway environmental exposures

Affiliations

¹ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: adurra@ucla.edu.
² UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; UCLA Environmental and Molecular Toxicology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
⁴ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; UCLA Environmental and Molecular Toxicology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA 90095, USA; Eli and Edythe Broad Stem Cell Research Center, UCLA, Los Angeles, CA 90095, USA; Division of Pulmonary and Critical Care Medicine, Department of Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA. Electronic address: bgomperts@mednet.ucla.edu.

PMID: 40924579
PMCID: PMC12451164
DOI: 10.1016/j.xpro.2025.104069

Protocol for constructing an accessible exposure chamber for in vitro and in vivo modeling of airway environmental exposures

Abdo Durra et al. STAR Protoc. 2025.

. 2025 Sep 19;6(3):104069.

doi: 10.1016/j.xpro.2025.104069. Epub 2025 Sep 8.

Affiliations

¹ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: adurra@ucla.edu.
² UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; UCLA Environmental and Molecular Toxicology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
⁴ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital, Department of Pediatrics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; UCLA Environmental and Molecular Toxicology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA 90095, USA; Eli and Edythe Broad Stem Cell Research Center, UCLA, Los Angeles, CA 90095, USA; Division of Pulmonary and Critical Care Medicine, Department of Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA. Electronic address: bgomperts@mednet.ucla.edu.

PMID: 40924579
PMCID: PMC12451164
DOI: 10.1016/j.xpro.2025.104069

Abstract

Exposure systems to study the effects of environmental exposures can be costly to purchase and difficult to use. Here, we present an accessible and cost-effective approach to building an exposure chamber in the lab. We describe steps for constructing the exposure system and writing the code to run it and simple instructions for experiments using the system. The exposure chamber can be adapted to expose airway cells or lab mice to combustible cigarette smoke, electronic cigarette vapors, or woodfire smoke. For complete details on the use and execution of this protocol, please refer to Durra et al.¹.

Keywords: Biotechnology and bioengineering; environmental sciences; health Sciences.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests.

Figures

**Figure 1**
Arduino IDE screenshots (A) Screenshot showing Arduino IDE software. (B) Screenshot showing Arduino board manager for installing the board package. (C) Screenshot showing Arduino IDE board selector.

**Figure 2**
Arduino and relay shield components and circuit schematics (A) A photo of an Arduino uno r4 with a red box indicating the USB-c port. (B) A photo of the Arduino 4 relay shield with red boxes indicating reset button, status LEDs, and relay pins. (C) Circuit schematic for Arduino control system for 1-pump operation. (D) Circuit schematic for Arduino control system for 2-pump operation.

**Figure 3**
Assembly steps for Arduino control system for 1-pump operation (A) A photo showing Arduino board with Arduino relay shield stacked on top of it. (B) A photo showing the female barrel jack adapter and a stripped 2-conductor cable wires. (C) A photo showing the female barrel jack adapter and 2-conductor cable wires connected to it. (D) A photo showing stripped wires in a loop next to the terminal block. (E) A photo showing the wires connected under the screws of the terminal block. (F) A photo showing stripped wires next to relay 1 pins of the Arduino relay shield. (G) A photo showing wires connected to the relay pins. (H) A photo showing 2-conductor cable wires connected to terminal block across wires with matching colors across. (I) A photo showing the other end of the 2-conductor cable wires with the female connector attached. (J) A photo showing the cable attached to the pump while matching the red wire to the pin with the red dot. (K) A photo showing the barrel jack of the power adapter plugged into the female barrel jack adapter with the wires connected to it. (L) A photo showing stripped wires in a loop next to the terminal block. (M) A photo showing the wires connected under the screws of the terminal block with jumper. (N) A photo showing stripped red wires next to the pins of relay 1 and 2 of the Arduino relay shield. (O) A photo showing a stripped blue wire next to the pins of relay 1 and 2 of the Arduino relay shield. (P) A photo showing connected to the pins of relay 1 and 2 of the Arduino relay shield. (Q) A photo showing connected to the pins of relay 1 and 2 of the Arduino relay shield. (R) A photo showing stripped wires in a loop next to the terminal block. (S) A photo showing the wires connected under the screws of the terminal block.

**Figure 4**
Preparing exposure material and setting up the exposure chamber (A) A schematic showing a combustible cigarette attached to the exposure pump. (B) A schematic showing a woodfire cone attached to the exposure pump. (C) A schematic showing an electronic cigarette attached to the exposure pump. (D) A schematic showing the exposure chamber for 1-pump operation of tissue culture plate exposure. (E) A schematic showing the exposure chamber for 2-pump operation of mice exposure. (F) An image showing the Arduino, exposure pump, Juul device with adapter, and exposure chamber inlet port.

**Figure 5**
Effects of combustible cigarettes, electronic cigarettes, and woodfire smoke on air-liquid interface cultures and mouse submucosal glands (A) Representative IF images of Keratin 5 (KRT5)-expressing (magenta) ABSCs in ALI cultures after exposure to combustible cigarettes smoke (CS), unflavored (vegetable glycerin propylene glycol 30:70 ratio with 5% nicotine) and mint flavored electronic cigarettes (e-cigs) vapors, and room air control. Scale bar: 100 μm. (B) Representative IF images of Keratin 5 (KRT5)-expressing (magenta) ABSCs in ALI cultures after exposure to woodfire smoke, and room air control. Scale bar: 50 μm. (C) Representative H&E staining of mouse trachea following exposure to combustible cigarettes smoke (CS), unflavored (vegetable glycerin propylene glycol 30:70 ratio with 5% nicotine). Scale bar: 100 μm.

See this image and copyright information in PMC

References

1. Durra A., Cherry C., Luo C., Hou E., Frauenpreis A., Purkayastha A., Passamano I., Makanani S., Castillo K., Lund A., et al. Unflavored electronic cigarette exposure induces alterations in airway ciliary structure and function. Respir. Res. 2025;26:223. doi: 10.1186/s12931-025-03302-w. - DOI - PMC - PubMed
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1. Zhang S., Gao Z., Wu L., Zhong Y., Gao H., Tao F.B., Wu X. Global patterns of asthma burden related to environmental risk factors during 1990-2019: an age-period-cohort analysis for global burden of disease study 2019. Environ. Health. 2024;23:20. doi: 10.1186/s12940-024-01060-8. - DOI - PMC - PubMed

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Protocol for constructing an accessible exposure chamber for in vitro and in vivo modeling of airway environmental exposures

Affiliations

Protocol for constructing an accessible exposure chamber for in vitro and in vivo modeling of airway environmental exposures

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

MeSH terms

Substances

LinkOut - more resources

Full Text Sources

Miscellaneous