Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix

Abstract Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evalu...

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Autores principales:	Manal Matoug-Elwerfelli, Hani Nazzal, El Mostafa Raif, Stacy-Paul Wilshaw, Filomena Esteves, Monty Duggal
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3c36b9e537374e85ad81f73898f35501
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spelling	oai:doaj.org-article:3c36b9e537374e85ad81f73898f355012021-12-02T11:43:35ZEx-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix10.1038/s41598-020-78477-x2045-2322https://doaj.org/article/3c36b9e537374e85ad81f73898f355012020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78477-xhttps://doaj.org/toc/2045-2322Abstract Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. Rat dental pulps were retrieved and divided into control and decellularised groups. The decellularisation protocol incorporated a low detergent concentration and hypotonic buffers. After decellularisation, the scaffolds were characterised histologically, immunohistochemistry and the residual DNA content quantified. Surface topography was also viewed under scanning electron microscopy. Biocompatibility was evaluated using cytotoxicity assays utilising L-929 cell line. Decellularised scaffolds were recellularised with human dental pulp stem cells up to 14 days in vitro. Cellular viability was assessed using LIVE/DEAD stain kit and the recellularised scaffolds were further assessed histologically and immunolabelled using makers for odontoblastic differentiation, cytoskeleton components and growth factors. Analysis of the decellularised scaffolds revealed an acellular matrix with histological preservation of structural components. Decellularised scaffolds were biocompatible and able to support stem cell survival following recellularisation. Immunolabelling of the recellularised scaffolds demonstrated positive cellular expression against the tested markers in culture. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Clinically, decellularised pulp tissue could possibly be a suitable scaffold for use within regenerative (reparative) endodontic techniques.Manal Matoug-ElwerfelliHani NazzalEl Mostafa RaifStacy-Paul WilshawFilomena EstevesMonty DuggalNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Manal Matoug-Elwerfelli Hani Nazzal El Mostafa Raif Stacy-Paul Wilshaw Filomena Esteves Monty Duggal Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
description	Abstract Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. Rat dental pulps were retrieved and divided into control and decellularised groups. The decellularisation protocol incorporated a low detergent concentration and hypotonic buffers. After decellularisation, the scaffolds were characterised histologically, immunohistochemistry and the residual DNA content quantified. Surface topography was also viewed under scanning electron microscopy. Biocompatibility was evaluated using cytotoxicity assays utilising L-929 cell line. Decellularised scaffolds were recellularised with human dental pulp stem cells up to 14 days in vitro. Cellular viability was assessed using LIVE/DEAD stain kit and the recellularised scaffolds were further assessed histologically and immunolabelled using makers for odontoblastic differentiation, cytoskeleton components and growth factors. Analysis of the decellularised scaffolds revealed an acellular matrix with histological preservation of structural components. Decellularised scaffolds were biocompatible and able to support stem cell survival following recellularisation. Immunolabelling of the recellularised scaffolds demonstrated positive cellular expression against the tested markers in culture. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Clinically, decellularised pulp tissue could possibly be a suitable scaffold for use within regenerative (reparative) endodontic techniques.
format	article
author	Manal Matoug-Elwerfelli Hani Nazzal El Mostafa Raif Stacy-Paul Wilshaw Filomena Esteves Monty Duggal
author_facet	Manal Matoug-Elwerfelli Hani Nazzal El Mostafa Raif Stacy-Paul Wilshaw Filomena Esteves Monty Duggal
author_sort	Manal Matoug-Elwerfelli
title	Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
title_short	Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
title_full	Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
title_fullStr	Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
title_full_unstemmed	Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
title_sort	ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/3c36b9e537374e85ad81f73898f35501
work_keys_str_mv	AT manalmatougelwerfelli exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix AT haninazzal exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix AT elmostafaraif exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix AT stacypaulwilshaw exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix AT filomenaesteves exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix AT montyduggal exvivorecellularisationandstemcelldifferentiationofadecellularisedratdentalpulpmatrix
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Ex-vivo recellularisation and stem cell differentiation of a decellularised rat dental pulp matrix

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