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Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype
To test the hypothesis that physical disruption of an intervertebral disc disturbs cell-matrix binding, leading to cell clustering and increased expression of matrix degrading enzymes that contribute towards degenerative disc cell phenotype. Lumbar disc tissue was removed at surgery from 21 patients...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6917743/ https://www.ncbi.nlm.nih.gov/pubmed/31871771 http://dx.doi.org/10.1038/s41420-019-0233-z |
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author | Lama, Polly Claireaux, Harry Flower, Luke Harding, Ian J. Dolan, Trish Le Maitre, Christine L. Adams, Michael A. |
author_facet | Lama, Polly Claireaux, Harry Flower, Luke Harding, Ian J. Dolan, Trish Le Maitre, Christine L. Adams, Michael A. |
author_sort | Lama, Polly |
collection | PubMed |
description | To test the hypothesis that physical disruption of an intervertebral disc disturbs cell-matrix binding, leading to cell clustering and increased expression of matrix degrading enzymes that contribute towards degenerative disc cell phenotype. Lumbar disc tissue was removed at surgery from 21 patients with disc herniation, 11 with disc degeneration, and 8 with adolescent scoliosis. 5 μm sections were examined with histology, and 30-µm sections by confocal microscopy. Antibodies were used against integrin α5beta1, matrix metalloproteinases (MMP) 1, MMP-3, caspase 3, and denatured collagen types I and II. Spatial associations were sought between cell clustering and various degenerative features. An additional, 11 non-herniated human discs were used to examine causality: half of each specimen was cultured in a manner that allowed free ‘unconstrained’ swelling (similar to a herniated disc in vivo), while the other half was cultured within a perspex ring that allowed ‘constrained’ swelling. Changes were monitored over 36 h using live-cell imaging. 1,9-Di-methyl methylene blue (DMMB) assay for glycosaminoglycan loss was carried out from tissue medium. Partially constrained specimens showed little swelling or cell movement in vitro. In contrast, unconstrained swelling significantly increased matrix distortion, glycosaminoglycan loss, exposure of integrin binding sites, expression of MMPs 1 and 3, and collagen denaturation. In the association studies, herniated disc specimens showed changes that resembled unconstrained swelling in vitro. In addition, they exhibited increased cell clustering, apoptosis, MMP expression, and collagen denaturation compared to ‘control’ discs. Results support our hypothesis. Further confirmation will require longitudinal animal experiments. |
format | Online Article Text |
id | pubmed-6917743 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69177432019-12-23 Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype Lama, Polly Claireaux, Harry Flower, Luke Harding, Ian J. Dolan, Trish Le Maitre, Christine L. Adams, Michael A. Cell Death Discov Article To test the hypothesis that physical disruption of an intervertebral disc disturbs cell-matrix binding, leading to cell clustering and increased expression of matrix degrading enzymes that contribute towards degenerative disc cell phenotype. Lumbar disc tissue was removed at surgery from 21 patients with disc herniation, 11 with disc degeneration, and 8 with adolescent scoliosis. 5 μm sections were examined with histology, and 30-µm sections by confocal microscopy. Antibodies were used against integrin α5beta1, matrix metalloproteinases (MMP) 1, MMP-3, caspase 3, and denatured collagen types I and II. Spatial associations were sought between cell clustering and various degenerative features. An additional, 11 non-herniated human discs were used to examine causality: half of each specimen was cultured in a manner that allowed free ‘unconstrained’ swelling (similar to a herniated disc in vivo), while the other half was cultured within a perspex ring that allowed ‘constrained’ swelling. Changes were monitored over 36 h using live-cell imaging. 1,9-Di-methyl methylene blue (DMMB) assay for glycosaminoglycan loss was carried out from tissue medium. Partially constrained specimens showed little swelling or cell movement in vitro. In contrast, unconstrained swelling significantly increased matrix distortion, glycosaminoglycan loss, exposure of integrin binding sites, expression of MMPs 1 and 3, and collagen denaturation. In the association studies, herniated disc specimens showed changes that resembled unconstrained swelling in vitro. In addition, they exhibited increased cell clustering, apoptosis, MMP expression, and collagen denaturation compared to ‘control’ discs. Results support our hypothesis. Further confirmation will require longitudinal animal experiments. Nature Publishing Group UK 2019-12-17 /pmc/articles/PMC6917743/ /pubmed/31871771 http://dx.doi.org/10.1038/s41420-019-0233-z Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Lama, Polly Claireaux, Harry Flower, Luke Harding, Ian J. Dolan, Trish Le Maitre, Christine L. Adams, Michael A. Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title | Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title_full | Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title_fullStr | Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title_full_unstemmed | Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title_short | Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
title_sort | physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6917743/ https://www.ncbi.nlm.nih.gov/pubmed/31871771 http://dx.doi.org/10.1038/s41420-019-0233-z |
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