Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding.
Hepatic tissue engineering using decellularized scaffolds is a potential therapeutic alternative to conventional transplantation. However, scaffolds are usually obtained using decellularization protocols that destroy the extracellular matrix (ECM) and hamper clinical translation. We aim to develop a...
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Public Library of Science (PLoS)
2016-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0155324&type=printable |
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| author | Panagiotis Maghsoudlou Fanourios Georgiades Holly Smith Anna Milan Panicos Shangaris Luca Urbani Stavros P Loukogeorgakis Benedetta Lombardi Giuseppe Mazza Charlotte Hagen Neil J Sebire Mark Turmaine Simon Eaton Alessandro Olivo Jasminka Godovac-Zimmermann Massimo Pinzani Paul Gissen Paolo De Coppi |
| author_facet | Panagiotis Maghsoudlou Fanourios Georgiades Holly Smith Anna Milan Panicos Shangaris Luca Urbani Stavros P Loukogeorgakis Benedetta Lombardi Giuseppe Mazza Charlotte Hagen Neil J Sebire Mark Turmaine Simon Eaton Alessandro Olivo Jasminka Godovac-Zimmermann Massimo Pinzani Paul Gissen Paolo De Coppi |
| author_sort | Panagiotis Maghsoudlou |
| collection | DOAJ |
| description | Hepatic tissue engineering using decellularized scaffolds is a potential therapeutic alternative to conventional transplantation. However, scaffolds are usually obtained using decellularization protocols that destroy the extracellular matrix (ECM) and hamper clinical translation. We aim to develop a decellularization technique that reliably maintains hepatic microarchitecture and ECM components. Isolated rat livers were decellularized by detergent-enzymatic technique with (EDTA-DET) or without EDTA (DET). Histology, DNA quantification and proteomics confirmed decellularization with further DNA reduction with the addition of EDTA. Quantification, histology, immunostaining, and proteomics demonstrated preservation of extracellular matrix components in both scaffolds with a higher amount of collagen and glycosaminoglycans in the EDTA-DET scaffold. Scanning electron microscopy and X-ray phase contrast imaging showed microarchitecture preservation, with EDTA-DET scaffolds more tightly packed. DET scaffold seeding with a hepatocellular cell line demonstrated complete repopulation in 14 days, with cells proliferating at that time. Decellularization using DET preserves microarchitecture and extracellular matrix components whilst allowing for cell growth for up to 14 days. Addition of EDTA creates a denser, more compact matrix. Transplantation of the scaffolds and scaling up of the methodology are the next steps for successful hepatic tissue engineering. |
| format | Article |
| id | doaj-art-8ca1f84e12334e29b8d1e72eb620114c |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-8ca1f84e12334e29b8d1e72eb620114c2025-08-20T02:03:13ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01115e015532410.1371/journal.pone.0155324Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding.Panagiotis MaghsoudlouFanourios GeorgiadesHolly SmithAnna MilanPanicos ShangarisLuca UrbaniStavros P LoukogeorgakisBenedetta LombardiGiuseppe MazzaCharlotte HagenNeil J SebireMark TurmaineSimon EatonAlessandro OlivoJasminka Godovac-ZimmermannMassimo PinzaniPaul GissenPaolo De CoppiHepatic tissue engineering using decellularized scaffolds is a potential therapeutic alternative to conventional transplantation. However, scaffolds are usually obtained using decellularization protocols that destroy the extracellular matrix (ECM) and hamper clinical translation. We aim to develop a decellularization technique that reliably maintains hepatic microarchitecture and ECM components. Isolated rat livers were decellularized by detergent-enzymatic technique with (EDTA-DET) or without EDTA (DET). Histology, DNA quantification and proteomics confirmed decellularization with further DNA reduction with the addition of EDTA. Quantification, histology, immunostaining, and proteomics demonstrated preservation of extracellular matrix components in both scaffolds with a higher amount of collagen and glycosaminoglycans in the EDTA-DET scaffold. Scanning electron microscopy and X-ray phase contrast imaging showed microarchitecture preservation, with EDTA-DET scaffolds more tightly packed. DET scaffold seeding with a hepatocellular cell line demonstrated complete repopulation in 14 days, with cells proliferating at that time. Decellularization using DET preserves microarchitecture and extracellular matrix components whilst allowing for cell growth for up to 14 days. Addition of EDTA creates a denser, more compact matrix. Transplantation of the scaffolds and scaling up of the methodology are the next steps for successful hepatic tissue engineering.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0155324&type=printable |
| spellingShingle | Panagiotis Maghsoudlou Fanourios Georgiades Holly Smith Anna Milan Panicos Shangaris Luca Urbani Stavros P Loukogeorgakis Benedetta Lombardi Giuseppe Mazza Charlotte Hagen Neil J Sebire Mark Turmaine Simon Eaton Alessandro Olivo Jasminka Godovac-Zimmermann Massimo Pinzani Paul Gissen Paolo De Coppi Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. PLoS ONE |
| title | Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. |
| title_full | Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. |
| title_fullStr | Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. |
| title_full_unstemmed | Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. |
| title_short | Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding. |
| title_sort | optimization of liver decellularization maintains extracellular matrix micro architecture and composition predisposing to effective cell seeding |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0155324&type=printable |
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