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Conference Paper: Production of a natural disc scaffold for intervertebral disc tissue engineering

TitleProduction of a natural disc scaffold for intervertebral disc tissue engineering
Authors
KeywordsMedical sciences
Orthopedics and traumatology
Issue Date2010
PublisherLippincott, Williams & Wilkins.
Citation
The 2010 Annual Meeting of the International Society for the Study of the Lumbar Spine (ISSLS), Auckland, New Zealand, 13-17 April 2010. In Spine: Affiliated Society Meeting Abstracts, October 2010, p. 192, Gp50 How to Cite?
AbstractINTRODUCTION: The physiological properties of the intervertebral disc (IVD) are highly linked to the composition of the extracellular matrix (ECM), of which current synthetic scaffolds lack a natural niche for cell growth. Decellularization helps to removecellular contents from a tissue whilst minimizing adverse effects on the ECM. This study aims to develop a natural disc scaffold with preserved ECM for disc cell culture by decellularization. METHODS: Bovine caudal discs (18‐23 mm in diameter) were harvested. Discs were incubated with 25 mL of phosphate buffer solution containing 0.1% sodium dodecyl sulphate (SDS) and protease inhibitor with constant agitation. Optimum conditions were identified using 4 different protocols which tested the effects of the duration and temperature of washing, frequency of solution replacement and number of snap‐freezing cycles during IVD decellularization. Live/Dead staining and the Alamar Blue assay were used for analysing the number of remaining cells and their metabolic activity after treatment. RESULTS: The most effective protocol enabled removal of 69% and 73% cell content from the annulus fibrosus and nucleus pulposus, respectively. Data from Alamar Blue assays suggested that the majority of the cell activity was eliminated. DISCUSSION: Increase of the duration of washing, frequency of solution replacement, and number of snap‐freezing cycles, decreased cell viability and facilitated removal of cellular contents. In this study, decellularization was achieved by physical treatment (snap‐freezing cycles) and chemical washing (SDS). Our findings imply that formation of intracellular ice crystals during the cycles facilitated the disruption of the cell membrane which allowed disc cell contents to be washed out. SDS buffer has a role in solubilizing cytoplasmic and nuclear membranes which aided in the clearance of cell remnants. Overall, this study shows that varying the frequency of solution change and duration of washing can effectively eliminate the majority of disc cells from the IVD. © 2010 Lippincott Williams & Wilkins, Inc.
DescriptionGeneral Posters: no. Gp50
Persistent Identifierhttp://hdl.handle.net/10722/126555
ISSN
2015 Impact Factor: 2.439
2015 SCImago Journal Rankings: 1.459

 

DC FieldValueLanguage
dc.contributor.authorChan, KYen_HK
dc.contributor.authorLeung, VYLen_HK
dc.contributor.authorTam, Ven_HK
dc.contributor.authorLu, WWen_HK
dc.contributor.authorSze, KYen_HK
dc.contributor.authorCheung, KMCen_HK
dc.date.accessioned2010-10-31T12:35:20Z-
dc.date.available2010-10-31T12:35:20Z-
dc.date.issued2010en_HK
dc.identifier.citationThe 2010 Annual Meeting of the International Society for the Study of the Lumbar Spine (ISSLS), Auckland, New Zealand, 13-17 April 2010. In Spine: Affiliated Society Meeting Abstracts, October 2010, p. 192, Gp50en_HK
dc.identifier.issn0362-2436-
dc.identifier.urihttp://hdl.handle.net/10722/126555-
dc.descriptionGeneral Posters: no. Gp50-
dc.description.abstractINTRODUCTION: The physiological properties of the intervertebral disc (IVD) are highly linked to the composition of the extracellular matrix (ECM), of which current synthetic scaffolds lack a natural niche for cell growth. Decellularization helps to removecellular contents from a tissue whilst minimizing adverse effects on the ECM. This study aims to develop a natural disc scaffold with preserved ECM for disc cell culture by decellularization. METHODS: Bovine caudal discs (18‐23 mm in diameter) were harvested. Discs were incubated with 25 mL of phosphate buffer solution containing 0.1% sodium dodecyl sulphate (SDS) and protease inhibitor with constant agitation. Optimum conditions were identified using 4 different protocols which tested the effects of the duration and temperature of washing, frequency of solution replacement and number of snap‐freezing cycles during IVD decellularization. Live/Dead staining and the Alamar Blue assay were used for analysing the number of remaining cells and their metabolic activity after treatment. RESULTS: The most effective protocol enabled removal of 69% and 73% cell content from the annulus fibrosus and nucleus pulposus, respectively. Data from Alamar Blue assays suggested that the majority of the cell activity was eliminated. DISCUSSION: Increase of the duration of washing, frequency of solution replacement, and number of snap‐freezing cycles, decreased cell viability and facilitated removal of cellular contents. In this study, decellularization was achieved by physical treatment (snap‐freezing cycles) and chemical washing (SDS). Our findings imply that formation of intracellular ice crystals during the cycles facilitated the disruption of the cell membrane which allowed disc cell contents to be washed out. SDS buffer has a role in solubilizing cytoplasmic and nuclear membranes which aided in the clearance of cell remnants. Overall, this study shows that varying the frequency of solution change and duration of washing can effectively eliminate the majority of disc cells from the IVD. © 2010 Lippincott Williams & Wilkins, Inc.-
dc.languageengen_HK
dc.publisherLippincott, Williams & Wilkins.-
dc.relation.ispartofSpine: Affiliated Society Meeting Abstracts-
dc.subjectMedical sciences-
dc.subjectOrthopedics and traumatology-
dc.titleProduction of a natural disc scaffold for intervertebral disc tissue engineeringen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailChan, KY: h0301351@hkusua.hku.hken_HK
dc.identifier.emailLeung, VYL: vicleung@hku.hken_HK
dc.identifier.emailTam, V: vivtam@hku.hken_HK
dc.identifier.emailLu, WW: wwlu@hku.hken_HK
dc.identifier.emailSze, KY: kysze@hku.hken_HK
dc.identifier.emailCheung, KMC: cheungmc@hku.hk-
dc.identifier.authorityLu, WW=rp00411en_HK
dc.identifier.authoritySze, KY=rp00171en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.hkuros180116en_HK
dc.identifier.volumeOctober 2010-
dc.identifier.spage192-
dc.identifier.epage192-
dc.publisher.placeUnited States-
dc.description.otherThe 2010 Annual Meeting of the International Society for the Study of the Lumbar Spine (ISSLS), Auckland, New Zealand, 13-17 April 2010. In Spine: Affiliated Society Meeting Abstracts, October 2010, p. 192, Gp50-

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