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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| Format: | Article |
| Language: | English |
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MDPI AG
2018-09-01
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| Series: | Journal of Functional Biomaterials |
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| Online Access: | http://www.mdpi.com/2079-4983/9/3/53 |
| _version_ | 1818036304508616704 |
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| author | Laurence Cantin-Warren Rina Guignard Sergio Cortez Ghio Danielle Larouche François A. Auger Lucie Germain |
| author_facet | Laurence Cantin-Warren Rina Guignard Sergio Cortez Ghio Danielle Larouche François A. Auger Lucie Germain |
| author_sort | Laurence Cantin-Warren |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-10T07:08:49Z |
| format | Article |
| id | doaj.art-a30eed627a6e44f096b5eb074c01aaf4 |
| institution | Directory Open Access Journal |
| issn | 2079-4983 |
| language | English |
| last_indexed | 2024-12-10T07:08:49Z |
| publishDate | 2018-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Functional Biomaterials |
| spelling | doaj.art-a30eed627a6e44f096b5eb074c01aaf42022-12-22T01:58:07ZengMDPI AGJournal of Functional Biomaterials2079-49832018-09-01935310.3390/jfb9030053jfb9030053Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue EngineeringLaurence Cantin-Warren0Rina Guignard1Sergio Cortez Ghio2Danielle Larouche3François A. Auger4Lucie Germain5Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaCentre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaCentre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaCentre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaCentre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaCentre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Département de Chirurgie, Faculté de Médecine, Université Laval, 1401 18e Rue, Québec, Québec G1J 1Z4, CanadaThere 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.http://www.mdpi.com/2079-4983/9/3/53culture techniquesregenerative medicineskin equivalenttissue culturebilayered skin substitutestissue engineeringskin ulcer |
| spellingShingle | Laurence Cantin-Warren Rina Guignard Sergio Cortez Ghio Danielle Larouche François A. Auger Lucie Germain Specialized Living Wound Dressing Based on the Self-Assembly Approach of Tissue Engineering Journal of Functional Biomaterials culture techniques regenerative medicine skin equivalent tissue culture bilayered skin substitutes tissue engineering skin ulcer |
| 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 | culture techniques regenerative medicine skin equivalent tissue culture bilayered skin substitutes tissue engineering skin ulcer |
| url | http://www.mdpi.com/2079-4983/9/3/53 |
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