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Conference Paper: Scaffold-free Three-dimensional Spheroids of DPSCs/ECs in Vascularised Pulp Regeneration

TitleScaffold-free Three-dimensional Spheroids of DPSCs/ECs in Vascularised Pulp Regeneration
Authors
KeywordsEndodontics
Odontoblasts
Pulp
Regeneration and Tissue engineering
Issue Date2013
PublisherSage Publications, Inc. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925
Citation
The 2nd Meeting of the International Association of Dental Research - Asia Pacific Region (IADR-APR), Bangkok, Thailand, 21-23 August 2013. In Journal of Dental Research, 2013, v. 92 n. Special Issue B: abstract no. 358 How to Cite?
AbstractObjectives: To determine the interactions between dental pulp stem cells (DPSCs) and endothelial cells (ECs) in a three-dimensional co-culture model; and to examine the potential of these cells in vascularised pulp regeneration. Methods: Three-dimensional microtissue-spheroids of DPSC-alone and DPSC-EC were fabricated using 12-series micro-molds (MicroTissues Inc.). Cells’ organization within the spheroids (CellTracker dyes) and viability (live/dead assay) were assessed at day-1, 7 and 14. Microtissue-spheroids 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 prevascularisation (Immunohistochemistry for CD31). Microtissues were inserted into the canal space of tooth-root fragments and implanted into the subcutaneous space on the back of 6- 8-week-old female severe combined immunodeficient (SCID) mice. 2-4 weeks after the transplantation, the mice were euthanized and the tooth fragments were removed for histological (Haematoxylin and eosin) and immunohistochemical (human mitochondria, CD31, DSP) analysis. Experiments were conducted in triplicate using DPSCs from three different donors and statistically analysed (ANOVA). Results: DPSCs and ECs were self-aggregated into spheroids with no evidence of cell death at the centre. ECs were organized into a dense-network of tubular-like structures throughout the DPSC:EC-microtissues. Elevated levels of ALP, BSP and RUNX2 (p < 0.05) in DPSC:EC-microtissues compared to DPSC-alone-microtissues confirmed that ECs enhanced osteo/odontogenic differentiation in 3D. Both DPSC-alone and DPSC:EC groups showed vascularised pulp-like tissue with an odontoblast-like cell layer adjacent to the dentin after transplantation in SCID mice. DPSC-EC-microtissue groups showed a significantly higher amount of extracellular matrix, vascularisation and mineralization compared to DPSC-alone-microtissues both in-vitro and in-vivo. Positive staining for antibodies against human mitochondria confirmed the contribution of transplanted microtissues in regenerated pulp-like tissue and vasculature. Conclusion: DPSCs and ECs synergistically enhance osteo/odontogenic differentiation and vascularisation in 3D in-vitro and regenerate vascularised pulp-like tissue in-vivo.
DescriptionConference Theme: We are the Future
Oral Presentation
Session 13: O-SEA Unilever competition (Senior)
Persistent Identifierhttp://hdl.handle.net/10722/192568
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorDissanayaka, WLen_US
dc.contributor.authorHargreaves, KMen_US
dc.contributor.authorJin, Len_US
dc.contributor.authorZhang, Cen_US
dc.date.accessioned2013-11-18T05:06:42Z-
dc.date.available2013-11-18T05:06:42Z-
dc.date.issued2013en_US
dc.identifier.citationThe 2nd Meeting of the International Association of Dental Research - Asia Pacific Region (IADR-APR), Bangkok, Thailand, 21-23 August 2013. In Journal of Dental Research, 2013, v. 92 n. Special Issue B: abstract no. 358en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/192568-
dc.descriptionConference Theme: We are the Future-
dc.descriptionOral Presentation-
dc.descriptionSession 13: O-SEA Unilever competition (Senior)-
dc.description.abstractObjectives: To determine the interactions between dental pulp stem cells (DPSCs) and endothelial cells (ECs) in a three-dimensional co-culture model; and to examine the potential of these cells in vascularised pulp regeneration. Methods: Three-dimensional microtissue-spheroids of DPSC-alone and DPSC-EC were fabricated using 12-series micro-molds (MicroTissues Inc.). Cells’ organization within the spheroids (CellTracker dyes) and viability (live/dead assay) were assessed at day-1, 7 and 14. Microtissue-spheroids 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 prevascularisation (Immunohistochemistry for CD31). Microtissues were inserted into the canal space of tooth-root fragments and implanted into the subcutaneous space on the back of 6- 8-week-old female severe combined immunodeficient (SCID) mice. 2-4 weeks after the transplantation, the mice were euthanized and the tooth fragments were removed for histological (Haematoxylin and eosin) and immunohistochemical (human mitochondria, CD31, DSP) analysis. Experiments were conducted in triplicate using DPSCs from three different donors and statistically analysed (ANOVA). Results: DPSCs and ECs were self-aggregated into spheroids with no evidence of cell death at the centre. ECs were organized into a dense-network of tubular-like structures throughout the DPSC:EC-microtissues. Elevated levels of ALP, BSP and RUNX2 (p < 0.05) in DPSC:EC-microtissues compared to DPSC-alone-microtissues confirmed that ECs enhanced osteo/odontogenic differentiation in 3D. Both DPSC-alone and DPSC:EC groups showed vascularised pulp-like tissue with an odontoblast-like cell layer adjacent to the dentin after transplantation in SCID mice. DPSC-EC-microtissue groups showed a significantly higher amount of extracellular matrix, vascularisation and mineralization compared to DPSC-alone-microtissues both in-vitro and in-vivo. Positive staining for antibodies against human mitochondria confirmed the contribution of transplanted microtissues in regenerated pulp-like tissue and vasculature. Conclusion: DPSCs and ECs synergistically enhance osteo/odontogenic differentiation and vascularisation in 3D in-vitro and regenerate vascularised pulp-like tissue in-vivo.-
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.subjectEndodontics-
dc.subjectOdontoblasts-
dc.subjectPulp-
dc.subjectRegeneration and Tissue engineering-
dc.titleScaffold-free Three-dimensional Spheroids of DPSCs/ECs in Vascularised Pulp Regenerationen_US
dc.typeConference_Paperen_US
dc.identifier.emailJin, L: ljjin@hkucc.hku.hken_US
dc.identifier.emailZhang, C: zhangcf@hku.hken_US
dc.identifier.authorityJin, L=rp00028en_US
dc.identifier.authorityZhang, C=rp01408en_US
dc.identifier.hkuros226798en_US
dc.identifier.volume92en_US
dc.identifier.issueSpecial Issue B: abstract no. 358en_US
dc.publisher.placeUnited States-

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