Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture
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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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-03-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1360089/full |
| _version_ | 1797261810055774208 |
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| author | Thomas Harry Jovic Thomas Harry Jovic Feihu Zhao Henry Jia Shareen Heather Doak Iain Stuart Whitaker Iain Stuart Whitaker |
| author_facet | Thomas Harry Jovic Thomas Harry Jovic Feihu Zhao Henry Jia Shareen Heather Doak Iain Stuart Whitaker Iain Stuart Whitaker |
| author_sort | Thomas Harry Jovic |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-24T23:47:08Z |
| format | Article |
| id | doaj.art-e92507eca901456c8f6e6062789d59f7 |
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| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-24T23:47:08Z |
| publishDate | 2024-03-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-e92507eca901456c8f6e6062789d59f72024-03-15T04:52:33ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852024-03-011210.3389/fbioe.2024.13600891360089Orbital shaking conditions augment human nasoseptal cartilage formation in 3D cultureThomas Harry Jovic0Thomas Harry Jovic1Feihu Zhao2Henry Jia3Shareen Heather Doak4Iain Stuart Whitaker5Iain Stuart Whitaker6Reconstructive Surgery & Regenerative Medicine Research Centre, Swansea University, Swansea, United KingdomWelsh Centre for Burns & Plastic Surgery, Morriston Hospital, Swansea, United KingdomDepartment of Biomedical Engineering & Zienkiewicz Institute, Faculty of Science and Engineering, Swansea University, Swansea, United KingdomReconstructive Surgery & Regenerative Medicine Research Centre, Swansea University, Swansea, United KingdomSwansea University Medical School, Swansea University, Swansea, United KingdomReconstructive Surgery & Regenerative Medicine Research Centre, Swansea University, Swansea, United KingdomWelsh Centre for Burns & Plastic Surgery, Morriston Hospital, Swansea, United KingdomIntroduction: 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.https://www.frontiersin.org/articles/10.3389/fbioe.2024.1360089/fullcartilagetissue engineeringchondrogenesisdynamic culturecomputational modelling |
| spellingShingle | Thomas Harry Jovic Thomas Harry Jovic Feihu Zhao Henry Jia Shareen Heather Doak Iain Stuart Whitaker Iain Stuart Whitaker Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture Frontiers in Bioengineering and Biotechnology cartilage tissue engineering chondrogenesis dynamic culture computational modelling |
| title | 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_short | Orbital shaking conditions augment human nasoseptal cartilage formation in 3D culture |
| title_sort | orbital shaking conditions augment human nasoseptal cartilage formation in 3d culture |
| topic | cartilage tissue engineering chondrogenesis dynamic culture computational modelling |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1360089/full |
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