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Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial...

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Autores principales: Van den Bogerd, Bert, Ní Dhubhghaill, Sorcha, Zakaria, Nadia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5947733/
https://www.ncbi.nlm.nih.gov/pubmed/29430874
http://dx.doi.org/10.1002/term.2633
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author Van den Bogerd, Bert
Ní Dhubhghaill, Sorcha
Zakaria, Nadia
author_facet Van den Bogerd, Bert
Ní Dhubhghaill, Sorcha
Zakaria, Nadia
author_sort Van den Bogerd, Bert
collection PubMed
description The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at −80 °C, and decellularized with water and trypsin‐EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell‐scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin‐EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds.
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spelling pubmed-59477332018-05-17 Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering Van den Bogerd, Bert Ní Dhubhghaill, Sorcha Zakaria, Nadia J Tissue Eng Regen Med Research Articles The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at −80 °C, and decellularized with water and trypsin‐EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell‐scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin‐EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds. John Wiley and Sons Inc. 2018-02-11 2018-04 /pmc/articles/PMC5947733/ /pubmed/29430874 http://dx.doi.org/10.1002/term.2633 Text en © 2017 The Authors Journal of Tissue Engineering and Regenerative Medicine Published by John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Van den Bogerd, Bert
Ní Dhubhghaill, Sorcha
Zakaria, Nadia
Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title_full Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title_fullStr Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title_full_unstemmed Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title_short Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
title_sort characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5947733/
https://www.ncbi.nlm.nih.gov/pubmed/29430874
http://dx.doi.org/10.1002/term.2633
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