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Conference Paper: ECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Model

TitleECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Model
Authors
KeywordsDentin
Endodontics
Pulp
Regeneration and Tissue engineering
Issue Date2012
PublisherSage Publications, Inc.. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925
Citation
The Annual Meeting of the International Association for Dental Research (IADR) Southeast Asian Division, Hong Kong, China, 3-4 November 2012. In Journal of Dental Research, 2012, v. 91 n. Special Issue C: abstract no. 168954 How to Cite?
AbstractObjectives: Dental pulp stem cells (DPSCs) are known to occupy a perivascular niche and interact with endothelial cells (ECs). This study, for the first time aimed to determine the interactions between DPSCs and ECs in a three-dimensional co-culture model, focusing on survival, angiogenesis and differentiation capacities to reflect cells’ in-vivo milieu more accurately. Methods: Three-dimensional microtissue-spheroids of DPSCs and ECs were fabricated using 12-series micro-molds (MicroTissues Inc.). CellTracker dyes were used to fluorescent label the cells and examined for the organization into spheroids. The cell viability was assessed with the live/dead viability assay kit at day-1, 7 and 14. Microtissue-spheroids (1200) were transferred to a custom-designed, 3mm-diameter, agarose mold and cultivated for 4-days to self-assemble into macrotissue. The macrotissues were induced for odontogenic differentiation (21-days), examined for expression levels of osteo/odontogenic markers: alkaline phosphatase (ALP), bone sialoprotein (BSP) and RUNX2 (Real-time PCR), mineralization (Von-Kossa) and for vascularisation (Immunohistochemistry for CD31). Experiments were conducted in triplicate using DPSCs from three different donors and statistically analysed (ANOVA). Results: DPSCs were aggregated to form spheroids when cultured with/without ECs in 3D conditions. The cell viability and turnover on day-14 remained equivalent to that of day-7 with no evidence of cell death in the centre of the spheroids. In contrast to DPSC-alone macrotissues, a dense-network of ECs was found throughout the DPSC:EC macrotissues under immunohistochemical analysis for the EC-specific marker CD31. Results confirmed that ECs enhanced osteo/odontogenic differentiation, on days-7 and 14, compared to DPSC only controls as shown by elevated ALP, BSP and RUNX2 levels (p < 0.05). DPSC-EC macrotissues showed a significantly higher amount of extracellular matrix and mineralization compared to DPSC-alone macrotissues in 3-D. Conclusions: ECs regulate DPSC activity and their differentiation capacity in 3D, which may facilitate the maintenance of DPSC quiescence and osteo/odontogenic differentiation when exposed to induction stimuli.
DescriptionSession: Pulp Biology and Regeneration
Persistent Identifierhttp://hdl.handle.net/10722/182062
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorDissanayaka, WLen_US
dc.contributor.authorZhang, Cen_US
dc.contributor.authorJin, Len_US
dc.contributor.authorHargreaves, KMen_US
dc.date.accessioned2013-04-17T07:20:43Z-
dc.date.available2013-04-17T07:20:43Z-
dc.date.issued2012en_US
dc.identifier.citationThe Annual Meeting of the International Association for Dental Research (IADR) Southeast Asian Division, Hong Kong, China, 3-4 November 2012. In Journal of Dental Research, 2012, v. 91 n. Special Issue C: abstract no. 168954en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/182062-
dc.descriptionSession: Pulp Biology and Regeneration-
dc.description.abstractObjectives: Dental pulp stem cells (DPSCs) are known to occupy a perivascular niche and interact with endothelial cells (ECs). This study, for the first time aimed to determine the interactions between DPSCs and ECs in a three-dimensional co-culture model, focusing on survival, angiogenesis and differentiation capacities to reflect cells’ in-vivo milieu more accurately. Methods: Three-dimensional microtissue-spheroids of DPSCs and ECs were fabricated using 12-series micro-molds (MicroTissues Inc.). CellTracker dyes were used to fluorescent label the cells and examined for the organization into spheroids. The cell viability was assessed with the live/dead viability assay kit at day-1, 7 and 14. Microtissue-spheroids (1200) were transferred to a custom-designed, 3mm-diameter, agarose mold and cultivated for 4-days to self-assemble into macrotissue. The macrotissues were induced for odontogenic differentiation (21-days), examined for expression levels of osteo/odontogenic markers: alkaline phosphatase (ALP), bone sialoprotein (BSP) and RUNX2 (Real-time PCR), mineralization (Von-Kossa) and for vascularisation (Immunohistochemistry for CD31). Experiments were conducted in triplicate using DPSCs from three different donors and statistically analysed (ANOVA). Results: DPSCs were aggregated to form spheroids when cultured with/without ECs in 3D conditions. The cell viability and turnover on day-14 remained equivalent to that of day-7 with no evidence of cell death in the centre of the spheroids. In contrast to DPSC-alone macrotissues, a dense-network of ECs was found throughout the DPSC:EC macrotissues under immunohistochemical analysis for the EC-specific marker CD31. Results confirmed that ECs enhanced osteo/odontogenic differentiation, on days-7 and 14, compared to DPSC only controls as shown by elevated ALP, BSP and RUNX2 levels (p < 0.05). DPSC-EC macrotissues showed a significantly higher amount of extracellular matrix and mineralization compared to DPSC-alone macrotissues in 3-D. Conclusions: ECs regulate DPSC activity and their differentiation capacity in 3D, which may facilitate the maintenance of DPSC quiescence and osteo/odontogenic differentiation when exposed to induction stimuli.-
dc.languageengen_US
dc.publisherSage Publications, Inc.. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925-
dc.relation.ispartofJournal of Dental Researchen_US
dc.rightsJournal of Dental Research. Copyright © Sage Publications, Inc..-
dc.subjectDentin-
dc.subjectEndodontics-
dc.subjectPulp-
dc.subjectRegeneration and Tissue engineering-
dc.titleECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Modelen_US
dc.typeConference_Paperen_US
dc.identifier.emailZhang, C: zhangcf@hku.hken_US
dc.identifier.emailJin, L: ljjin@hkucc.hku.hken_US
dc.identifier.authorityZhang, C=rp01408en_US
dc.identifier.authorityJin, L=rp00028en_US
dc.identifier.hkuros213910en_US
dc.identifier.volume91en_US
dc.identifier.issueSpecial Issue C: abstract no. 168954en_US
dc.publisher.placeUnited States-

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