High-resolution 3D micro-CT reveals an additional muscle layer in esophageal atresia
9 affiliations
- 1Department of Paediatric Surgery, Emma Children’s Hospital, Amsterdam UMC, location AMC, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
- 2Department of Obstetrics and Gynaecology, Amsterdam UMC, location AMC, Meibergdreef 9, 1105AZ Amsterdam, The Netherlands
- 3Amsterdam Reproduction and Development Research Institute, Meibergdreef 9, 1105AZ Amsterdam, The Netherlands
- 4Amsterdam Gastroenterology Endocrinology Metabolism Research Institute, Meibergdreef 9, 1105AZ Amsterdam, The Netherlands
- 5Department of Medical Biology, Amsterdam UMC, location AMC, Meibergdreef 15, 1105AZ Amsterdam, The Netherlands
- 6Erasmus MC – Sophia Children’s Hospital, Department of Paediatric Surgery, University Medical Center Rotterdam, Rotterdam, The Netherlands
- 7Department of Pathology, Amsterdam UMC, location AMC, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
- 8Department of Computer Science, University of Copenhagen, Universitetsparken 1, 2100 København, Denmark
- 9Department of Paediatric Gastroenterology, Emma Children’s Hospital, Amsterdam UMC, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Abstract
Background: Esophageal atresia (EA) is a congenital malformation intrinsically associated with esophageal dysmotility. Its etiology is poorly understood and it is unclear why motility is better preserved in some patients than in others. Esophageal motility is a three-dimensional (3D) process orchestrated by microscale structures, but conventional histology offers two-dimensional insight in the organization of the esophageal wall, hindering the identification of subtle, but potentially relevant, microarchitectural abnormalities that may contribute to understanding of the etiology of EA and variability in motility patterns. We aimed to evaluate the 3D microscale architecture of the esophagus in EA patients and identify potential anomalies using microfocus computed tomography (micro-CT).
Methods: Full thickness esophageal samples from the proximal and distal pouch of eleven neonates undergoing EA repair and three fetal control esophagus samples (gestational age: 24 weeks) were collected, contrast-enhanced, and imaged with high-resolution micro-CT (2–5 μm voxel size). Muscle layers were segmented using the machine learning software RootPainter and EA samples were compared with the fetal controls. Findings were validated by histology.
Results: Micro-CT enabled clear visualization of epithelial, submucosal, vascular, and muscular structures in all samples. 3D reconstructions consistently revealed a previously unreported oblique muscle layer situated medial to the circular muscle layer in all EA specimens. This layer varied in prominence and was not recognizable on 2D histology alone. Fetal control samples displayed only the typical circular and longitudinal layers, with oblique muscle fibres confined to the stomach.
Conclusion: Three-dimensional assessment of esophageal microarchitecture using micro-CT allows differentiation between epithelium, musculature and vascularization. Moreover, it facilitated the identification of an oblique muscle layer in proximal pouch and distal trachea-esophageal fistula specimens from EA patients. It remains to be studied whether these findings can explain the observed variation in motility patterns in EA patients.
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BibTeX
@article{docter2026highresolution,
title = {{High-resolution 3D micro-CT reveals an additional muscle layer in esophageal atresia}},
author = {Daniel Docter and Anastasia Ioannidou and Bernadette de Bakker and Joris J. T. H. Roelofs and Sjoerd de Beer and Sander Zwaveling and Jermo Hanemaaijer-van der Veer and Jon Sporring and Joep Derikx and Matthijs Oomen and Michiel van Wijk and Ramon Gorter},
journal = {Esophagus},
year = {2026},
month = aug,
doi = {10.1007/s10388-026-01242-y},
url = {https://doi.org/10.1007/s10388-026-01242-y},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - High-resolution 3D micro-CT reveals an additional muscle layer in esophageal atresia
AU - Daniel Docter
AU - Anastasia Ioannidou
AU - Bernadette de Bakker
AU - Joris J. T. H. Roelofs
AU - Sjoerd de Beer
AU - Sander Zwaveling
AU - Jermo Hanemaaijer-van der Veer
AU - Jon Sporring
AU - Joep Derikx
AU - Matthijs Oomen
AU - Michiel van Wijk
AU - Ramon Gorter
PY - 2026
DA - 2026/08/31
JO - Esophagus
DO - 10.1007/s10388-026-01242-y
UR - https://doi.org/10.1007/s10388-026-01242-y
AB - Background: Esophageal atresia (EA) is a congenital malformation intrinsically associated with esophageal dysmotility. Its etiology is poorly understood and it is unclear why motility is better preserved in some patients than in others. Esophageal motility is a three-dimensional (3D) process orchestrated by microscale structures, but conventional histology offers two-dimensional insight in the organization of the esophageal wall, hindering the identification of subtle, but potentially relevant, microarchitectural abnormalities that may contribute to understanding of the etiology of EA and variability in motility patterns. We aimed to evaluate the 3D microscale architecture of the esophagus in EA patients and identify potential anomalies using microfocus computed tomography (micro-CT). Methods: Full thickness esophageal samples from the proximal and distal pouch of eleven neonates undergoing EA repair and three fetal control esophagus samples (gestational age: 24 weeks) were collected, contrast-enhanced, and imaged with high-resolution micro-CT (2–5 μm voxel size). Muscle layers were segmented using the machine learning software RootPainter and EA samples were compared with the fetal controls. Findings were validated by histology. Results: Micro-CT enabled clear visualization of epithelial, submucosal, vascular, and muscular structures in all samples. 3D reconstructions consistently revealed a previously unreported oblique muscle layer situated medial to the circular muscle layer in all EA specimens. This layer varied in prominence and was not recognizable on 2D histology alone. Fetal control samples displayed only the typical circular and longitudinal layers, with oblique muscle fibres confined to the stomach. Conclusion: Three-dimensional assessment of esophageal microarchitecture using micro-CT allows differentiation between epithelium, musculature and vascularization. Moreover, it facilitated the identification of an oblique muscle layer in proximal pouch and distal trachea-esophageal fistula specimens from EA patients. It remains to be studied whether these findings can explain the observed variation in motility patterns in EA patients.
SN - 1612-9059
SN - 1612-9067
ER -