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Amelioration of Subglottic Stenosis by Antimicrobial Peptide Eluting Endotracheal Tubes

  • SI: 2023 CMBE Young Innovators
  • Published:
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Abstract

Introduction

Pediatric subglottic stenosis (SGS) results from prolonged intubation where scar tissue leads to airway narrowing that requires invasive surgery. We have recently discovered that modulating the laryngotracheal microbiome can prevent SGS. Herein, we show how our patent-pending antimicrobial peptide-eluting endotracheal tube (AMP-ET) effectively modulates the local airway microbiota resulting in reduced inflammation and stenosis resolution.

Materials and Methods

We fabricated mouse-sized ETs coated with a polymeric AMP-eluting layer, quantified AMP release over 10 days, and validated bactericidal activity for both planktonic and biofilm-resident bacteria against Staphylococcus aureus and Pseudomonas aeruginosa. Ex vivo testing: we inserted AMP-ETs and ET controls into excised laryngotracheal complexes (LTCs) of C57BL/6 mice and assessed biofilm formation after 24 h. In vivo testing: AMP-ETs and ET controls were inserted in sham or SGS-induced LTCs, which were then implanted subcutaneously in receptor mice, and assessed for immune response and SGS severity after 7 days.

Results

We achieved reproducible, linear AMP release at 1.16 µg/day resulting in strong bacterial inhibition in vitro and ex vivo. In vivo, SGS-induced LTCs exhibited a thickened scar tissue typical of stenosis, while the use of AMP-ETs abrogated stenosis. Notably, SGS airways exhibited high infiltration of T cells and macrophages, which was reversed with AMP-ET treatment. This suggests that by modulating the microbiome, AMP-ETs reduce macrophage activation and antigen specific T cell responses resolving stenosis progression.

Conclusion

We developed an AMP-ET platform that reduces T cell and macrophage responses and reduces SGS in vivo via airway microbiome modulation.

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Data availability

Data is available upon request.

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Acknowledgements

We thank the Microbial Culture and Metabolomics Core of the PennCHOP Microbiome Program (Philadelphia, PA) for bacterial cultures and Dr. Susan Thibeault for kindly providing the vocal folds fibroblasts used in this study. We also thank Kyra Smith for her help with the graphical illustrations. This work was carried out in part at the Singh Center for Nanotechnology, which is supported by the NSF National Nanotechnology Coordinated Infrastructure Program under Grant NNCI-2025608. This work was supported in part by the Children’s Hospital of Philadelphia Research Institute (RG), the Frontier Program in Airway Disorders of the Children’s Hospital of Philadelphia (RG), Foerderer Grant (RG), and the National Science Foundation Graduate Research Fellowship No. DGE 1845298 (MRA, RF).

Author information

Authors and Affiliations

  1. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

    Matthew R. Aronson, Amrita Mehta, Ryan M. Friedman, Daniel D. Ghaderi & Riccardo Gottardi

  2. Division of Otolaryngology, Department of Surgery, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    Matthew R. Aronson, Amrita Mehta, Ryan M. Friedman, Daniel D. Ghaderi, Ryan C. Borek, Ian N. Jacobs & Riccardo Gottardi

  3. Division of Otolaryngology, Philadelphia Veterans Affairs Medical Center, Philadelphia, PA, USA

    Matthew R. Aronson, Hoang C. B. Nguyen & Natasha Mirza

  4. Department of Otorhinolaryngology-Head and Neck Surgery, University of Pennsylvania, Philadelphia, PA, USA

    Ryan C. Borek, Hoang C. B. Nguyen, Ian N. Jacobs, Natasha Mirza & Riccardo Gottardi

  5. Department of Laboratory Animal Services, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    Kendra S. McDaid

  6. Division of Pulmonary Medicine, Department of Pediatrics, University of Pennsylvania, Philadelphia, PA, USA

    Riccardo Gottardi

  7. Department of Orthopaedic Surgery, University of Pennsylvania, Philadelphia, PA, USA

    Riccardo Gottardi

  8. Ri.MED Foundation, Palermo, Italy

    Riccardo Gottardi

  9. Children’s Hospital of Philadelphia, Abramson Research Center, 3615 Civic Center Boulevard, Room 1006, Philadelphia, PA, 19107, USA

    Riccardo Gottardi

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Contributions

MRA, RCB, INJ, NM, and RG conceived the research plan. MRA, AM, RMF, DDG, HCBN, and KSM carried out experiments. MRA, RMF, and RG prepared figures and wrote the manuscript. All authors reviewed, edited, and approved the final manuscript.

Corresponding author

Correspondence to Riccardo Gottardi.

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Conflict of interest

MA and RG are inventors on a pending patent related to the technology described in this manuscript. AM, RMF, DDG, RCB, HCBN, KSM, INJ, and NM declare no conflicts of interest.

Research Involving Human and Animal Rights Statement

All institutional and national guidelines for the care and use of laboratory animals were followed and approved by the appropriate institutional committees. No human studies were carried out by the authors for this article.

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Associate Editor Owen McCarty oversaw the review of this article.

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Aronson, M.R., Mehta, A., Friedman, R.M. et al. Amelioration of Subglottic Stenosis by Antimicrobial Peptide Eluting Endotracheal Tubes. Cel. Mol. Bioeng. 16, 369–381 (2023). https://doi.org/10.1007/s12195-023-00769-9

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