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Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture

Thomas Jovic, Feihu Zhao Orcid Logo, Henry Jia, Shareen Doak Orcid Logo, Iain Whitaker

Frontiers in Bioengineering and Biotechnology, Volume: 12

Swansea University Authors: Thomas Jovic, Feihu Zhao Orcid Logo, Henry Jia, Shareen Doak Orcid Logo, Iain Whitaker

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DOI (Published version): 10.3389/fbioe.2024.1360089

Abstract

Introduction: This study aimed to determine whether a dynamic orbital shaking culture system could enhance the cartilage production and viability of bioengineered nasoseptal cartilage.Methods: Human nasal chondrocytes were seeded onto nanocellulose-alginate biomaterials and cultured in static or dyn...

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Published in: Frontiers in Bioengineering and Biotechnology
ISSN: 2296-4185
Published: Frontiers Media SA 2024
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URI: https://cronfa.swan.ac.uk/Record/cronfa68089
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Quantitative polymerase chain reaction for chondrogenic gene expression (type 2 collagen, aggrecan and SOX9) was performed, demonstrating a transient rise in SOX9 expression at 1 and 7 days of culture, followed by a rise at 7 and 14 days in Aggrecan (184.5-fold increase, p &lt; 0.0001) and Type 2 Collagen (226.3-fold increase, p = 0.049) expression. Samples were analysed histologically for glycosaminoglycan content using Alcian blue staining and demonstrated increased matrix formation in dynamic culture.Results: Superior cell viability was identified in the dynamic conditions through live-dead and alamarBlue assays. Computational analysis was used to determine the shear stress experienced by cells in the biomaterial in the dynamic conditions and found that the mechanical stimulation exerted was minimal (fluid shear stress &lt;0.02 mPa, fluid pressure &lt;48 Pa).Conclusion: We conclude that the use of an orbital shaking system exerts biologically relevant effects on bioengineered nasoseptal cartilage independently of the expected thresholds of mechanical stimulation, with implications for optimising future cartilage tissue engineering efforts.</abstract><type>Journal Article</type><journal>Frontiers in Bioengineering and Biotechnology</journal><volume>12</volume><journalNumber/><paginationStart/><paginationEnd/><publisher>Frontiers Media SA</publisher><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint/><issnElectronic>2296-4185</issnElectronic><keywords/><publishedDay>15</publishedDay><publishedMonth>3</publishedMonth><publishedYear>2024</publishedYear><publishedDate>2024-03-15</publishedDate><doi>10.3389/fbioe.2024.1360089</doi><url>http://dx.doi.org/10.3389/fbioe.2024.1360089</url><notes/><college>COLLEGE NANME</college><department>Medical School</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>MEDS</DepartmentCode><institution>Swansea University</institution><apcterm/><funders>This study was supported by The Scar Free Foundation &amp; Health and Care Research Wales Programme of research in Reconstructive Surgery &amp; Regenerative Medicine, which has been established in the ReconRegen Research Centre at Swansea University in partnership with Swansea Bay University Health Board. This work was additionally supported by Action Medical Research and the VTCT Foundation (Grant Number 2782), the Royal College of Surgeons England and the Welsh Clinical Academic Training Programme.</funders><projectreference/><lastEdited>2024-10-29T09:34:30.0049281</lastEdited><Created>2024-10-29T09:30:41.0923299</Created><path><level id="1">Faculty of Science and Engineering</level><level id="2">School of Engineering and Applied Sciences - Biomedical Engineering</level></path><authors><author><firstname>Thomas</firstname><surname>Jovic</surname><order>1</order></author><author><firstname>Feihu</firstname><surname>Zhao</surname><orcid>0000-0003-0515-6808</orcid><order>2</order></author><author><firstname>Henry</firstname><surname>Jia</surname><order>3</order></author><author><firstname>Shareen</firstname><surname>Doak</surname><orcid>0000-0002-6753-1987</orcid><order>4</order></author><author><firstname>Iain</firstname><surname>Whitaker</surname><orcid/><order>5</order></author></authors><documents/><OutputDurs/></rfc1807>
spelling v2 68089 2024-10-29 Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture 7d95ed2bceb18fc0fdfd4048277c6eed Thomas Jovic Thomas Jovic true false 1c6e79b6edd08c88a8d17a241cd78630 0000-0003-0515-6808 Feihu Zhao Feihu Zhao true false b5967fb7820fe78c0770f71b5769369c Henry Jia Henry Jia true false 8f70286908f67238a527a98cbf66d387 0000-0002-6753-1987 Shareen Doak Shareen Doak true false 830074c59291938a55b480dcbee4697e Iain Whitaker Iain Whitaker true false 2024-10-29 MEDS Introduction: This study aimed to determine whether a dynamic orbital shaking culture system could enhance the cartilage production and viability of bioengineered nasoseptal cartilage.Methods: Human nasal chondrocytes were seeded onto nanocellulose-alginate biomaterials and cultured in static or dynamic conditions for 14 days. Quantitative polymerase chain reaction for chondrogenic gene expression (type 2 collagen, aggrecan and SOX9) was performed, demonstrating a transient rise in SOX9 expression at 1 and 7 days of culture, followed by a rise at 7 and 14 days in Aggrecan (184.5-fold increase, p < 0.0001) and Type 2 Collagen (226.3-fold increase, p = 0.049) expression. Samples were analysed histologically for glycosaminoglycan content using Alcian blue staining and demonstrated increased matrix formation in dynamic culture.Results: Superior cell viability was identified in the dynamic conditions through live-dead and alamarBlue assays. Computational analysis was used to determine the shear stress experienced by cells in the biomaterial in the dynamic conditions and found that the mechanical stimulation exerted was minimal (fluid shear stress <0.02 mPa, fluid pressure <48 Pa).Conclusion: We conclude that the use of an orbital shaking system exerts biologically relevant effects on bioengineered nasoseptal cartilage independently of the expected thresholds of mechanical stimulation, with implications for optimising future cartilage tissue engineering efforts. Journal Article Frontiers in Bioengineering and Biotechnology 12 Frontiers Media SA 2296-4185 15 3 2024 2024-03-15 10.3389/fbioe.2024.1360089 http://dx.doi.org/10.3389/fbioe.2024.1360089 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University This study was supported by The Scar Free Foundation & Health and Care Research Wales Programme of research in Reconstructive Surgery & Regenerative Medicine, which has been established in the ReconRegen Research Centre at Swansea University in partnership with Swansea Bay University Health Board. This work was additionally supported by Action Medical Research and the VTCT Foundation (Grant Number 2782), the Royal College of Surgeons England and the Welsh Clinical Academic Training Programme. 2024-10-29T09:34:30.0049281 2024-10-29T09:30:41.0923299 Faculty of Science and Engineering School of Engineering and Applied Sciences - Biomedical Engineering Thomas Jovic 1 Feihu Zhao 0000-0003-0515-6808 2 Henry Jia 3 Shareen Doak 0000-0002-6753-1987 4 Iain Whitaker 5
title Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
spellingShingle Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
Thomas Jovic
Feihu Zhao
Henry Jia
Shareen Doak
Iain Whitaker
title_short Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
title_full Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
title_fullStr Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
title_full_unstemmed Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
title_sort Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
author_id_str_mv 7d95ed2bceb18fc0fdfd4048277c6eed
1c6e79b6edd08c88a8d17a241cd78630
b5967fb7820fe78c0770f71b5769369c
8f70286908f67238a527a98cbf66d387
830074c59291938a55b480dcbee4697e
author_id_fullname_str_mv 7d95ed2bceb18fc0fdfd4048277c6eed_***_Thomas Jovic
1c6e79b6edd08c88a8d17a241cd78630_***_Feihu Zhao
b5967fb7820fe78c0770f71b5769369c_***_Henry Jia
8f70286908f67238a527a98cbf66d387_***_Shareen Doak
830074c59291938a55b480dcbee4697e_***_Iain Whitaker
author Thomas Jovic
Feihu Zhao
Henry Jia
Shareen Doak
Iain Whitaker
author2 Thomas Jovic
Feihu Zhao
Henry Jia
Shareen Doak
Iain Whitaker
format Journal article
container_title Frontiers in Bioengineering and Biotechnology
container_volume 12
publishDate 2024
institution Swansea University
issn 2296-4185
doi_str_mv 10.3389/fbioe.2024.1360089
publisher Frontiers Media SA
college_str Faculty of Science and Engineering
hierarchytype
hierarchy_top_id facultyofscienceandengineering
hierarchy_top_title Faculty of Science and Engineering
hierarchy_parent_id facultyofscienceandengineering
hierarchy_parent_title Faculty of Science and Engineering
department_str School of Engineering and Applied Sciences - Biomedical Engineering{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Biomedical Engineering
url http://dx.doi.org/10.3389/fbioe.2024.1360089
document_store_str 0
active_str 0
description Introduction: This study aimed to determine whether a dynamic orbital shaking culture system could enhance the cartilage production and viability of bioengineered nasoseptal cartilage.Methods: Human nasal chondrocytes were seeded onto nanocellulose-alginate biomaterials and cultured in static or dynamic conditions for 14 days. Quantitative polymerase chain reaction for chondrogenic gene expression (type 2 collagen, aggrecan and SOX9) was performed, demonstrating a transient rise in SOX9 expression at 1 and 7 days of culture, followed by a rise at 7 and 14 days in Aggrecan (184.5-fold increase, p < 0.0001) and Type 2 Collagen (226.3-fold increase, p = 0.049) expression. Samples were analysed histologically for glycosaminoglycan content using Alcian blue staining and demonstrated increased matrix formation in dynamic culture.Results: Superior cell viability was identified in the dynamic conditions through live-dead and alamarBlue assays. Computational analysis was used to determine the shear stress experienced by cells in the biomaterial in the dynamic conditions and found that the mechanical stimulation exerted was minimal (fluid shear stress <0.02 mPa, fluid pressure <48 Pa).Conclusion: We conclude that the use of an orbital shaking system exerts biologically relevant effects on bioengineered nasoseptal cartilage independently of the expected thresholds of mechanical stimulation, with implications for optimising future cartilage tissue engineering efforts.
published_date 2024-03-15T09:34:29Z
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score 11.0351515

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