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Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics
Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of thre...
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Published in: | European journal of orthodontics 2020-09, Vol.42 (4), p.387-395 |
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container_title | European journal of orthodontics |
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creator | Brezulier, Damien Pellen-Mussi, Pascal Tricot-Doleux, Sylvie Novella, Agnès Sorel, Olivier Jeanne, Sylvie |
description | Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of three-dimensional (3D) culture of osteoblasts to study mechanobiology.
The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription.
The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable.
The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days.
The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis. |
doi_str_mv | 10.1093/ejo/cjaa017 |
format | article |
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The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription.
The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable.
The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days.
The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis.</description><identifier>ISSN: 0141-5387</identifier><identifier>EISSN: 1460-2210</identifier><identifier>DOI: 10.1093/ejo/cjaa017</identifier><identifier>PMID: 32144430</identifier><language>eng</language><publisher>England: Oxford University Press (OUP)</publisher><subject>Chemical Sciences ; Dentistry</subject><ispartof>European journal of orthodontics, 2020-09, Vol.42 (4), p.387-395</ispartof><rights>The Author(s) 2020. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c360t-3d18a698aad10ccdfe68623a25ea545e009a5ddae9d1391fe9f0ffab684e9d603</citedby><cites>FETCH-LOGICAL-c360t-3d18a698aad10ccdfe68623a25ea545e009a5ddae9d1391fe9f0ffab684e9d603</cites><orcidid>0000-0002-1519-9943 ; 0000-0003-2185-282X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27922,27923</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32144430$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://univ-rennes.hal.science/hal-02533165$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Brezulier, Damien</creatorcontrib><creatorcontrib>Pellen-Mussi, Pascal</creatorcontrib><creatorcontrib>Tricot-Doleux, Sylvie</creatorcontrib><creatorcontrib>Novella, Agnès</creatorcontrib><creatorcontrib>Sorel, Olivier</creatorcontrib><creatorcontrib>Jeanne, Sylvie</creatorcontrib><title>Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics</title><title>European journal of orthodontics</title><addtitle>Eur J Orthod</addtitle><description>Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of three-dimensional (3D) culture of osteoblasts to study mechanobiology.
The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription.
The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable.
The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days.
The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis.</description><subject>Chemical Sciences</subject><subject>Dentistry</subject><issn>0141-5387</issn><issn>1460-2210</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNo9kU1LJDEQhoMoOrp72vuSo7K0VjpJfxxFXRUGvOyeQ01ScXpId2Y7aWH-vT3MrKeCl6ceqHoZ-yHgVkAr72gT7-wGEUR9whZCVVCUpYBTtgChRKFlU1-wy5Q2ACAbVZ-zC1kKpZSEBbOP9EEhbnsaMo-eI5ePfD31OPCYMsVVwJS5pRC4nUKeRuJ9dBS4jyNPeXK7bnjnPdk1DnHVxRDfd7ybl8e8ji4OubPpGzvzGBJ9P84r9vf305-Hl2L59vz6cL8srKwgF9KJBqu2QXQCrHWeqqYqJZaaUCtNAC1q55BaJ2QrPLUevMdV1ag5qkBesZuDd43BbMeux3FnInbm5X5p9hmUWkpR6Q8xs9cHdjvGfxOlbPou7c_EgeKUTClrJXVZ67321wG1Y0xpJP_lFmD2DZi5AXNsYKZ_HsXTqif3xf5_ufwEjuyDmw</recordid><startdate>20200911</startdate><enddate>20200911</enddate><creator>Brezulier, Damien</creator><creator>Pellen-Mussi, Pascal</creator><creator>Tricot-Doleux, Sylvie</creator><creator>Novella, Agnès</creator><creator>Sorel, Olivier</creator><creator>Jeanne, Sylvie</creator><general>Oxford University Press (OUP)</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><orcidid>https://orcid.org/0000-0002-1519-9943</orcidid><orcidid>https://orcid.org/0000-0003-2185-282X</orcidid></search><sort><creationdate>20200911</creationdate><title>Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics</title><author>Brezulier, Damien ; Pellen-Mussi, Pascal ; Tricot-Doleux, Sylvie ; Novella, Agnès ; Sorel, Olivier ; Jeanne, Sylvie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c360t-3d18a698aad10ccdfe68623a25ea545e009a5ddae9d1391fe9f0ffab684e9d603</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Chemical Sciences</topic><topic>Dentistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Brezulier, Damien</creatorcontrib><creatorcontrib>Pellen-Mussi, Pascal</creatorcontrib><creatorcontrib>Tricot-Doleux, Sylvie</creatorcontrib><creatorcontrib>Novella, Agnès</creatorcontrib><creatorcontrib>Sorel, Olivier</creatorcontrib><creatorcontrib>Jeanne, Sylvie</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><jtitle>European journal of orthodontics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Brezulier, Damien</au><au>Pellen-Mussi, Pascal</au><au>Tricot-Doleux, Sylvie</au><au>Novella, Agnès</au><au>Sorel, Olivier</au><au>Jeanne, Sylvie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics</atitle><jtitle>European journal of orthodontics</jtitle><addtitle>Eur J Orthod</addtitle><date>2020-09-11</date><risdate>2020</risdate><volume>42</volume><issue>4</issue><spage>387</spage><epage>395</epage><pages>387-395</pages><issn>0141-5387</issn><eissn>1460-2210</eissn><abstract>Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of three-dimensional (3D) culture of osteoblasts to study mechanobiology.
The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription.
The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable.
The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days.
The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis.</abstract><cop>England</cop><pub>Oxford University Press (OUP)</pub><pmid>32144430</pmid><doi>10.1093/ejo/cjaa017</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-1519-9943</orcidid><orcidid>https://orcid.org/0000-0003-2185-282X</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Chemical Sciences Dentistry |
title | Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics |
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