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Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering

There is a high incidence of failure and recurrence for chronic skin wounds following conventional therapies. To promote healing, the use of skin substitutes containing living cells as wound dressings has been proposed. The aim of this study was to produce a scaffold-free cell-based bilayered tissue...

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Autores principales: Cantin-Warren, Laurence, Guignard, Rina, Cortez Ghio, Sergio, Larouche, Danielle, Auger, François A., Germain, Lucie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6165032/
https://www.ncbi.nlm.nih.gov/pubmed/30223550
http://dx.doi.org/10.3390/jfb9030053
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author Cantin-Warren, Laurence
Guignard, Rina
Cortez Ghio, Sergio
Larouche, Danielle
Auger, François A.
Germain, Lucie
author_facet Cantin-Warren, Laurence
Guignard, Rina
Cortez Ghio, Sergio
Larouche, Danielle
Auger, François A.
Germain, Lucie
author_sort Cantin-Warren, Laurence
collection PubMed
description There is a high incidence of failure and recurrence for chronic skin wounds following conventional therapies. To promote healing, the use of skin substitutes containing living cells as wound dressings has been proposed. The aim of this study was to produce a scaffold-free cell-based bilayered tissue-engineered skin substitute (TES) containing living fibroblasts and keratinocytes suitable for use as wound dressing, while considering production time, handling effort during the manufacturing process, and stability of the final product. The self-assembly method, which relies on the ability of mesenchymal cells to secrete and organize connective tissue sheet sustaining keratinocyte growth, was used to produce TESs. Three fibroblast-seeding densities were tested to produce tissue sheets. At day 17, keratinocytes were added onto 1 or 3 (reference method) stacked tissue sheets. Four days later, TESs were subjected either to 4, 10, or 17 days of culture at the air–liquid interface (A/L). All resulting TESs were comparable in terms of their histological aspect, protein expression profile and contractile behavior in vitro. However, signs of extracellular matrix (ECM) digestion that progressed over culture time were noted in TESs produced with only one fibroblast-derived tissue sheet. With lower fibroblast density, the ECM of TESs was almost completely digested after 10 days A/L and lost histological integrity after grafting in athymic mice. Increasing the fibroblast seeding density 5 to 10 times solved this problem. We conclude that the proposed method allows for a 25-day production of a living TES, which retains its histological characteristics in vitro for at least two weeks.
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spelling pubmed-61650322018-10-12 Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering Cantin-Warren, Laurence Guignard, Rina Cortez Ghio, Sergio Larouche, Danielle Auger, François A. Germain, Lucie J Funct Biomater Article There is a high incidence of failure and recurrence for chronic skin wounds following conventional therapies. To promote healing, the use of skin substitutes containing living cells as wound dressings has been proposed. The aim of this study was to produce a scaffold-free cell-based bilayered tissue-engineered skin substitute (TES) containing living fibroblasts and keratinocytes suitable for use as wound dressing, while considering production time, handling effort during the manufacturing process, and stability of the final product. The self-assembly method, which relies on the ability of mesenchymal cells to secrete and organize connective tissue sheet sustaining keratinocyte growth, was used to produce TESs. Three fibroblast-seeding densities were tested to produce tissue sheets. At day 17, keratinocytes were added onto 1 or 3 (reference method) stacked tissue sheets. Four days later, TESs were subjected either to 4, 10, or 17 days of culture at the air–liquid interface (A/L). All resulting TESs were comparable in terms of their histological aspect, protein expression profile and contractile behavior in vitro. However, signs of extracellular matrix (ECM) digestion that progressed over culture time were noted in TESs produced with only one fibroblast-derived tissue sheet. With lower fibroblast density, the ECM of TESs was almost completely digested after 10 days A/L and lost histological integrity after grafting in athymic mice. Increasing the fibroblast seeding density 5 to 10 times solved this problem. We conclude that the proposed method allows for a 25-day production of a living TES, which retains its histological characteristics in vitro for at least two weeks. MDPI 2018-09-15 /pmc/articles/PMC6165032/ /pubmed/30223550 http://dx.doi.org/10.3390/jfb9030053 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cantin-Warren, Laurence
Guignard, Rina
Cortez Ghio, Sergio
Larouche, Danielle
Auger, François A.
Germain, Lucie
Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title_full Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title_fullStr Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title_full_unstemmed Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title_short Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering
title_sort specialized living wound dressing based on the self-assembly approach of tissue engineering
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6165032/
https://www.ncbi.nlm.nih.gov/pubmed/30223550
http://dx.doi.org/10.3390/jfb9030053
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