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Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus
Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9551564/ https://www.ncbi.nlm.nih.gov/pubmed/36237211 http://dx.doi.org/10.3389/fbioe.2022.937239 |
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author | Herrera Quijano, Marco A. Sharma, Nadia Morissette Martin, Pascal Séguin, Cheryle A. Flynn, Lauren E. |
author_facet | Herrera Quijano, Marco A. Sharma, Nadia Morissette Martin, Pascal Séguin, Cheryle A. Flynn, Lauren E. |
author_sort | Herrera Quijano, Marco A. |
collection | PubMed |
description | Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of two-dimensional (2-D) and three-dimensional (3-D) cell culture platforms incorporating decellularized nucleus pulposus (DNP). First, a detergent-free protocol was developed for decellularizing bovine nucleus pulposus (NP) tissues that was effective at removing cellular content while preserving key ECM constituents including collagens, glycosaminoglycans, and the cell-adhesive glycoproteins laminin and fibronectin. Next, novel 2-D coatings were generated using the DNP or commercially-sourced bovine collagen type I (COL) as a non-tissue-specific control. In addition, cryo-milled DNP or COL particles were incorporated within methacrylated chondroitin sulphate (MCS) hydrogels as a 3-D cell culture platform for exploring the effects of ECM particle composition. Culture studies showed that the 2-D coatings derived from the DNP could support cell attachment and growth, but did not maintain or rescue the phenotype of primary bovine NP cells, which de-differentiated when serially passaged in monolayer culture. Similarly, while bovine NP cells remained highly viable following encapsulation and 14 days of culture within the hydrogel composites, the incorporation of DNP particles within the MCS hydrogels was insufficient to maintain or rescue changes in NP phenotype associated with extended in vitro culture based on gene expression patterns. Overall, DNP produced with our new decellularization protocol was successfully applied to generate both 2-D and 3-D bioscaffolds; however, further studies are required to assess if these platforms can be combined with additional components of the endogenous NP microenvironment to stimulate regeneration or lineage-specific cell differentiation. |
format | Online Article Text |
id | pubmed-9551564 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95515642022-10-12 Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus Herrera Quijano, Marco A. Sharma, Nadia Morissette Martin, Pascal Séguin, Cheryle A. Flynn, Lauren E. Front Bioeng Biotechnol Bioengineering and Biotechnology Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of two-dimensional (2-D) and three-dimensional (3-D) cell culture platforms incorporating decellularized nucleus pulposus (DNP). First, a detergent-free protocol was developed for decellularizing bovine nucleus pulposus (NP) tissues that was effective at removing cellular content while preserving key ECM constituents including collagens, glycosaminoglycans, and the cell-adhesive glycoproteins laminin and fibronectin. Next, novel 2-D coatings were generated using the DNP or commercially-sourced bovine collagen type I (COL) as a non-tissue-specific control. In addition, cryo-milled DNP or COL particles were incorporated within methacrylated chondroitin sulphate (MCS) hydrogels as a 3-D cell culture platform for exploring the effects of ECM particle composition. Culture studies showed that the 2-D coatings derived from the DNP could support cell attachment and growth, but did not maintain or rescue the phenotype of primary bovine NP cells, which de-differentiated when serially passaged in monolayer culture. Similarly, while bovine NP cells remained highly viable following encapsulation and 14 days of culture within the hydrogel composites, the incorporation of DNP particles within the MCS hydrogels was insufficient to maintain or rescue changes in NP phenotype associated with extended in vitro culture based on gene expression patterns. Overall, DNP produced with our new decellularization protocol was successfully applied to generate both 2-D and 3-D bioscaffolds; however, further studies are required to assess if these platforms can be combined with additional components of the endogenous NP microenvironment to stimulate regeneration or lineage-specific cell differentiation. Frontiers Media S.A. 2022-09-27 /pmc/articles/PMC9551564/ /pubmed/36237211 http://dx.doi.org/10.3389/fbioe.2022.937239 Text en Copyright © 2022 Herrera Quijano, Sharma, Morissette Martin, Séguin and Flynn. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Herrera Quijano, Marco A. Sharma, Nadia Morissette Martin, Pascal Séguin, Cheryle A. Flynn, Lauren E. Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title_full | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title_fullStr | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title_full_unstemmed | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title_short | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
title_sort | development of 2-d and 3-d culture platforms derived from decellularized nucleus pulposus |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9551564/ https://www.ncbi.nlm.nih.gov/pubmed/36237211 http://dx.doi.org/10.3389/fbioe.2022.937239 |
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