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Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents

Human plasma-derived bilayered skin substitutes were successfully used by our group to produce human-based in vitro skin models for toxicity, cosmetic, and pharmaceutical testing. However, mechanical weakness, which causes the plasma-derived fibrin matrices to contract significantly, led us to attem...

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Autores principales: Montero, Andrés, Quílez, Cristina, Valencia, Leticia, Girón, Paula, Jorcano, José Luis, Velasco, Diego
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8269027/
https://www.ncbi.nlm.nih.gov/pubmed/34201667
http://dx.doi.org/10.3390/ijms22136746
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author Montero, Andrés
Quílez, Cristina
Valencia, Leticia
Girón, Paula
Jorcano, José Luis
Velasco, Diego
author_facet Montero, Andrés
Quílez, Cristina
Valencia, Leticia
Girón, Paula
Jorcano, José Luis
Velasco, Diego
author_sort Montero, Andrés
collection PubMed
description Human plasma-derived bilayered skin substitutes were successfully used by our group to produce human-based in vitro skin models for toxicity, cosmetic, and pharmaceutical testing. However, mechanical weakness, which causes the plasma-derived fibrin matrices to contract significantly, led us to attempt to improve their stability. In this work, we studied whether an increase in fibrin concentration from 1.2 to 2.4 mg/mL (which is the useful fibrinogen concentration range that can be obtained from plasma) improves the matrix and, hence, the performance of the in vitro skin cultures. The results show that this increase in fibrin concentration indeed affected the mechanical properties by doubling the elastic moduli and the maximum load. A structural analysis indicated a decreased porosity for the 2.4 mg/mL hydrogels, which can help explain this mechanical behavior. The contraction was clearly reduced for the 2.4 mg/mL matrices, which also allowed for the growth and proliferation of primary fibroblasts and keratinocytes, although at a somewhat reduced rate compared to the 1.2 mg/mL gels. Finally, both concentrations of fibrin gave rise to organotypic skin cultures with a fully differentiated epidermis, although their lifespans were longer (25–35%) in cultures with more concentrated matrices, which improves their usefulness. These systems will allow the generation of much better in vitro skin models for the testing of drugs, cosmetics and chemicals, or even to “personalized” skin for the diagnosis or determination of the most effective treatment possible.
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spelling pubmed-82690272021-07-10 Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents Montero, Andrés Quílez, Cristina Valencia, Leticia Girón, Paula Jorcano, José Luis Velasco, Diego Int J Mol Sci Article Human plasma-derived bilayered skin substitutes were successfully used by our group to produce human-based in vitro skin models for toxicity, cosmetic, and pharmaceutical testing. However, mechanical weakness, which causes the plasma-derived fibrin matrices to contract significantly, led us to attempt to improve their stability. In this work, we studied whether an increase in fibrin concentration from 1.2 to 2.4 mg/mL (which is the useful fibrinogen concentration range that can be obtained from plasma) improves the matrix and, hence, the performance of the in vitro skin cultures. The results show that this increase in fibrin concentration indeed affected the mechanical properties by doubling the elastic moduli and the maximum load. A structural analysis indicated a decreased porosity for the 2.4 mg/mL hydrogels, which can help explain this mechanical behavior. The contraction was clearly reduced for the 2.4 mg/mL matrices, which also allowed for the growth and proliferation of primary fibroblasts and keratinocytes, although at a somewhat reduced rate compared to the 1.2 mg/mL gels. Finally, both concentrations of fibrin gave rise to organotypic skin cultures with a fully differentiated epidermis, although their lifespans were longer (25–35%) in cultures with more concentrated matrices, which improves their usefulness. These systems will allow the generation of much better in vitro skin models for the testing of drugs, cosmetics and chemicals, or even to “personalized” skin for the diagnosis or determination of the most effective treatment possible. MDPI 2021-06-23 /pmc/articles/PMC8269027/ /pubmed/34201667 http://dx.doi.org/10.3390/ijms22136746 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Montero, Andrés
Quílez, Cristina
Valencia, Leticia
Girón, Paula
Jorcano, José Luis
Velasco, Diego
Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title_full Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title_fullStr Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title_full_unstemmed Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title_short Effect of Fibrin Concentration on the In Vitro Production of Dermo-Epidermal Equivalents
title_sort effect of fibrin concentration on the in vitro production of dermo-epidermal equivalents
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8269027/
https://www.ncbi.nlm.nih.gov/pubmed/34201667
http://dx.doi.org/10.3390/ijms22136746
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