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Causative Role of Anoxic Environment in Bacterial Regulation of Human Intestinal Function

  • S.I. : 2022 CMBE Young Innovators
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Abstract

Introduction

Life on Earth depends on oxygen; human tissues require oxygen signaling, whereas many microorganisms, including bacteria, thrive in anoxic environments. Despite these differences, human tissues and bacteria coexist in close proximity to each other such as in the intestine. How oxygen governs intestinal-bacterial interactions remains poorly understood.

Methods

To address to this gap, we created a dual-oxygen environment in a microfluidic device to study the role of oxygen in regulating the regulation of intestinal enzymes and proteins by gut bacteria. Two-layer microfluidic devices were designed using a fluid transport model and fabricated using soft lithography. An oxygen-sensitive material was integrated to determine the oxygen levels. The intestinal cells were cultured in the upper chamber of the device. The cells were differentiated, upon which bacterial strains, a facultative anaerobe, Escherichia coli Nissle 1917, and an obligate anaerobe, Bifidobacterium Adolescentis, were cultured with the intestinal cells.

Results

The microfluidic device successfully established a dual-oxygen environment. Of particular importance in our findings was that both strains significantly upregulated mucin proteins and modulated several intestinal transporters and transcription factors but only under the anoxic–oxic oxygen gradient, thus providing evidence of the role of oxygen on bacterial-epithelial signaling.

Conclusions

Our work that integrates cell and molecular biology with bioengineering presents a novel strategy to engineer an accessible experimental system to provide tailored oxygenated environments. The work could provide new avenues to study intestine-microbiome signaling and intestinal tissue engineering, as well as a novel perspective on the indirect effects of gut bacteria on tissues including tumors.

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Data Availability

The data and material are listed in supplementary documents.

Code Availability

Not applicable.

Abbreviations

O2C−:

Culture condition with no control of oxygen

O2C+:

Culture condition with oxygen control

ECN:

E. coli Nissle 1917

Bifido :

Bifidobacterium adolescentis (B. adolescentis)

MOI:

Multiplicity of Infection of bacteria to cells

CYP:

Cytochrome P450

MUC2:

Mucin 2

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Funding

This work was partly funded by the Nayar Prize II, Alternatives Research and Development Foundation (ARDF) and student scholarships from the Armor College of Engineering.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Illinois Institute of Technology, 3255 S. Dearborn Street, Wishnick Hall, Chicago, IL, 60616, USA

    Chengyao Wang, Andrea Cancino, Jasmine Baste, Daniel Marten, Advait Anil Joshi, Amreen Nasreen & Abhinav Bhushan

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Contributions

Participated in research design: CW, AB. Conducted experiments: CW, AC, JB, DM, AAJ, AN. Contributed new reagents or analytic tools: CW, AB. Performed data analysis: CW, JB, AAJ, AB. Performed data visualization: CW, JB, AC, DM. Wrote or contributed to the writing of the manuscript: CW, AC, JB, DM, AB.

Corresponding author

Correspondence to Abhinav Bhushan.

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Conflict of interest

The authors Chengyao Wang, Andrea Cancino, Jasmine Baste, Daniel Marten, Advait Anil Joshi, Amreen Nasreen, Abhinav Bhushan have declared that no conflict of interest exists.

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Associate Editor Cheng Dong oversaw the review of this article.

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Wang, C., Cancino, A., Baste, J. et al. Causative Role of Anoxic Environment in Bacterial Regulation of Human Intestinal Function. Cel. Mol. Bioeng. 15, 493–504 (2022). https://doi.org/10.1007/s12195-022-00735-x

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