Cargando…

The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line

Bone is a dynamic organ that can adapt its structure to meet the demands of its biochemical and biophysical environment. Osteocytes form a sensory network throughout the tissue and orchestrate tissue adaptation via the release of soluble factors such as a sclerostin. Osteocyte physiology has traditi...

Descripción completa

Detalles Bibliográficos
Autores principales: Brady, Robert T., O’Brien, Fergal J., Hoey, David A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8772728/
https://www.ncbi.nlm.nih.gov/pubmed/35049744
http://dx.doi.org/10.3390/bioengineering9010035
_version_ 1784635911035682816
author Brady, Robert T.
O’Brien, Fergal J.
Hoey, David A.
author_facet Brady, Robert T.
O’Brien, Fergal J.
Hoey, David A.
author_sort Brady, Robert T.
collection PubMed
description Bone is a dynamic organ that can adapt its structure to meet the demands of its biochemical and biophysical environment. Osteocytes form a sensory network throughout the tissue and orchestrate tissue adaptation via the release of soluble factors such as a sclerostin. Osteocyte physiology has traditionally been challenging to investigate due to the uniquely mineralized extracellular matrix (ECM) of bone leading to the development of osteocyte cell lines. Importantly, the most widely researched and utilized osteocyte cell line: the MLO-Y4, is limited by its inability to express sclerostin (Sost gene) in typical in-vitro culture. We theorised that culture in an environment closer to the in vivo osteocyte environment could impact on Sost expression. Therefore, this study investigated the role of composition and dimensionality in directing Sost expression in MLO-Y4 cells using collagen-based ECM analogues. A significant outcome of this study is that MLO-Y4 cells, when cultured on a hydroxyapatite (HA)-containing two-dimensional (2D) film analogue, expressed Sost. Moreover, three-dimensional (3D) culture within HA-containing collagen scaffolds significantly enhanced Sost expression, demonstrating the impact of ECM composition and dimensionality on MLO-Y4 behaviour. Importantly, in this bone mimetic ECM environment, Sost expression was found to be comparable to physiological levels. Lastly, MLO-Y4 cells cultured in these novel conditions responded accordingly to fluid flow stimulation with a decrease in expression. This study therefore presents a novel culture system for the MLO-Y4 osteocyte cell line, ensuring the expression of an important osteocyte specific gene, Sost, overcoming a major limitation of this model.
format Online
Article
Text
id pubmed-8772728
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87727282022-01-21 The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line Brady, Robert T. O’Brien, Fergal J. Hoey, David A. Bioengineering (Basel) Article Bone is a dynamic organ that can adapt its structure to meet the demands of its biochemical and biophysical environment. Osteocytes form a sensory network throughout the tissue and orchestrate tissue adaptation via the release of soluble factors such as a sclerostin. Osteocyte physiology has traditionally been challenging to investigate due to the uniquely mineralized extracellular matrix (ECM) of bone leading to the development of osteocyte cell lines. Importantly, the most widely researched and utilized osteocyte cell line: the MLO-Y4, is limited by its inability to express sclerostin (Sost gene) in typical in-vitro culture. We theorised that culture in an environment closer to the in vivo osteocyte environment could impact on Sost expression. Therefore, this study investigated the role of composition and dimensionality in directing Sost expression in MLO-Y4 cells using collagen-based ECM analogues. A significant outcome of this study is that MLO-Y4 cells, when cultured on a hydroxyapatite (HA)-containing two-dimensional (2D) film analogue, expressed Sost. Moreover, three-dimensional (3D) culture within HA-containing collagen scaffolds significantly enhanced Sost expression, demonstrating the impact of ECM composition and dimensionality on MLO-Y4 behaviour. Importantly, in this bone mimetic ECM environment, Sost expression was found to be comparable to physiological levels. Lastly, MLO-Y4 cells cultured in these novel conditions responded accordingly to fluid flow stimulation with a decrease in expression. This study therefore presents a novel culture system for the MLO-Y4 osteocyte cell line, ensuring the expression of an important osteocyte specific gene, Sost, overcoming a major limitation of this model. MDPI 2022-01-13 /pmc/articles/PMC8772728/ /pubmed/35049744 http://dx.doi.org/10.3390/bioengineering9010035 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Brady, Robert T.
O’Brien, Fergal J.
Hoey, David A.
The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title_full The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title_fullStr The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title_full_unstemmed The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title_short The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line
title_sort impact of the extracellular matrix environment on sost expression by the mlo-y4 osteocyte cell line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8772728/
https://www.ncbi.nlm.nih.gov/pubmed/35049744
http://dx.doi.org/10.3390/bioengineering9010035
work_keys_str_mv AT bradyrobertt theimpactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline
AT obrienfergalj theimpactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline
AT hoeydavida theimpactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline
AT bradyrobertt impactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline
AT obrienfergalj impactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline
AT hoeydavida impactoftheextracellularmatrixenvironmentonsostexpressionbythemloy4osteocytecellline