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Detergent enzymatic treatment for the development of a natural acellular matrix for oesophageal regeneration

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Abstract

Purpose

Tissue engineering of the oesophagus has been proposed as a therapeutic alternative to organ transplantation. We previously demonstrated that a detergent enzymatic treatment (DET) is a valid method to obtain an acellular matrix with preservation of the native architecture. In this study, we aimed to develop a natural acellular matrix from pig oesophagus, as a valid framework for oesophageal replacement.

Methods

Pig oesophagi (n = 4) were decellularized with continuous luminal infusion of DET. To evaluate the efficiency of the decellularization, samples were assessed by histology and DNA quantification. Moreover, the ultra-structural characteristics of the acellular matrix were investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM).

Results

Decellularization of the oesophagus was achieved after three cycles of DET. Histological analysis showed the maintenance of tissue matrix architecture with absence of cellular elements, verified by measurement of DNA. SEM and TEM analysis confirmed preservation of the ultra-structural characteristics of the native tissue.

Conclusions

Oesophageal acellular matrix can be successfully obtained by decellularization of pig oesophagus using a gentle DET via the oesophageal lumen. This decellularization method preserves the ultrastructure of the native tissue and could represent the basis for a tissue-engineered oesophagus.

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Acknowledgments

We thank Michael Boyd of the Royal Veterinary College’s Biological Unit for providing the animal tissue. This investigation was supported by Great Ormond Street Hospital charity, the Fondation Eugenio Litta (Geneva, Switzerland), the Medical Research Council, the Royal College of Surgeons of England, the Sparks Children’s Medical Charity, the British Foreign Office for the UK/USA Stem Cell Collaboration Award and the Mittal Research Fund. We would also like to thank the Royal Society/Wolfson Foundation for the tissue engineering laboratory refurbishment grant obtained for the Pediatric Surgery Department in the Institute of Child Health.

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Authors and Affiliations

  1. Surgery Unit, Institute of Child Health and Great Ormond Street Hospital, University College London, 30 Guilford Street, London, WC1N 1EH, UK

    Giorgia Totonelli, Panagiotis Maghsoudlou, Fanourious Georgiades, Massimo Garriboli, Kiron Koshy, Agostino Pierro, Simon Eaton & Paolo De Coppi

  2. Division of Bioscience, University College London, London, WC1N 1EH, UK

    Mark Turmaine

  3. Department of Histopathology, Institute of Child Health and Great Ormond Street Hospital, University College London, London, WC1N 1EH, UK

    Michael Ashworth & Neil J. Sebire

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  1. Giorgia Totonelli
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  11. Paolo De Coppi
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Correspondence to Paolo De Coppi.

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Totonelli, G., Maghsoudlou, P., Georgiades, F. et al. Detergent enzymatic treatment for the development of a natural acellular matrix for oesophageal regeneration. Pediatr Surg Int 29, 87–95 (2013). https://doi.org/10.1007/s00383-012-3194-3

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