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Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets

This paper describes a novel method for the rapid construction of skin, using multiple layers of aligned electrospun fibers as starting scaffolds. Scaffolds were spun from biodegradable L-lactide/glycolide (molar ratio 10:90) with predominantly parallel arrays of fibers attached peripherally to thin...

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Autores principales: Merrilees, Mervyn, Buunk, Neil, Zuo, Ning, Larsen, Nigel, Karimi, Samaneh, Tucker, Nick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7825689/
https://www.ncbi.nlm.nih.gov/pubmed/33430199
http://dx.doi.org/10.3390/bioengineering8010007
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author Merrilees, Mervyn
Buunk, Neil
Zuo, Ning
Larsen, Nigel
Karimi, Samaneh
Tucker, Nick
author_facet Merrilees, Mervyn
Buunk, Neil
Zuo, Ning
Larsen, Nigel
Karimi, Samaneh
Tucker, Nick
author_sort Merrilees, Mervyn
collection PubMed
description This paper describes a novel method for the rapid construction of skin, using multiple layers of aligned electrospun fibers as starting scaffolds. Scaffolds were spun from biodegradable L-lactide/glycolide (molar ratio 10:90) with predominantly parallel arrays of fibers attached peripherally to thin 304 stainless steel layer frames. Each layer frame was held between two thicker support frames. Human skin cells were seeded onto multiple (three–nine) scaffolds. Dermal fibroblasts were seeded on both sides of each scaffold except for one on which keratinocytes were seeded on one side only. Following 48 h of culture, the scaffolds and layer frames were unmounted from their support frames, stacked, with keratinocytes uppermost, and securely held in place by upper and lower support frames to instantly form a multilayered “dermis” and a nascent epidermis. The stack was cultured for a further 5 days during which time the cells proliferated and then adhered to form, in association with the spun fibers, a mechanically coherent tissue. Fibroblasts preferentially elongated in the dominant fiber direction and a two-dimensional weave of alternating fiber and cell alignments could be constructed by selected placement of the layer frames during stacking. Histology of the 7-day tissue stacks showed the organized layers of fibroblasts and keratinocytes immuno-positive for keratin. Electron microscopy showed attachment of fibroblasts to the lactide/glycolide fibers and small-diameter collagen fibers in the extracellular space. This novel approach could be used to engineer a range of tissues for grafting where rapid construction of tissues with aligned or woven layers would be beneficial.
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spelling pubmed-78256892021-01-24 Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets Merrilees, Mervyn Buunk, Neil Zuo, Ning Larsen, Nigel Karimi, Samaneh Tucker, Nick Bioengineering (Basel) Article This paper describes a novel method for the rapid construction of skin, using multiple layers of aligned electrospun fibers as starting scaffolds. Scaffolds were spun from biodegradable L-lactide/glycolide (molar ratio 10:90) with predominantly parallel arrays of fibers attached peripherally to thin 304 stainless steel layer frames. Each layer frame was held between two thicker support frames. Human skin cells were seeded onto multiple (three–nine) scaffolds. Dermal fibroblasts were seeded on both sides of each scaffold except for one on which keratinocytes were seeded on one side only. Following 48 h of culture, the scaffolds and layer frames were unmounted from their support frames, stacked, with keratinocytes uppermost, and securely held in place by upper and lower support frames to instantly form a multilayered “dermis” and a nascent epidermis. The stack was cultured for a further 5 days during which time the cells proliferated and then adhered to form, in association with the spun fibers, a mechanically coherent tissue. Fibroblasts preferentially elongated in the dominant fiber direction and a two-dimensional weave of alternating fiber and cell alignments could be constructed by selected placement of the layer frames during stacking. Histology of the 7-day tissue stacks showed the organized layers of fibroblasts and keratinocytes immuno-positive for keratin. Electron microscopy showed attachment of fibroblasts to the lactide/glycolide fibers and small-diameter collagen fibers in the extracellular space. This novel approach could be used to engineer a range of tissues for grafting where rapid construction of tissues with aligned or woven layers would be beneficial. MDPI 2021-01-07 /pmc/articles/PMC7825689/ /pubmed/33430199 http://dx.doi.org/10.3390/bioengineering8010007 Text en © 2021 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
Merrilees, Mervyn
Buunk, Neil
Zuo, Ning
Larsen, Nigel
Karimi, Samaneh
Tucker, Nick
Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title_full Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title_fullStr Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title_full_unstemmed Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title_short Use of Stacked Layers of Electrospun L-Lactide/Glycolide Co-Polymer Fibers for Rapid Construction of Skin Sheets
title_sort use of stacked layers of electrospun l-lactide/glycolide co-polymer fibers for rapid construction of skin sheets
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7825689/
https://www.ncbi.nlm.nih.gov/pubmed/33430199
http://dx.doi.org/10.3390/bioengineering8010007
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