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Conference Paper: Prevascularized Three-Dimensional DPSC Constructs in Dental Pulp Regeneration

TitlePrevascularized Three-Dimensional DPSC Constructs in Dental Pulp Regeneration
Authors
KeywordsPulp
Regeneration and Stem Cells
Issue Date2014
PublisherSage Publications, Inc. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925
Citation
The 92nd General Session & Exhibition of the International Association for Dental Research (IADR), Cape Town, South Africa, 25-28 June 2014. In Journal of Dental Research, 2014, v. 93 n. Special issue B: abstract no. 174 How to Cite?
AbstractObjectives: To co-culture dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs) in an injectable, peptide hydrogel scaffold: PuraMatrix while fabricating a vascular network in-vitro and to utilize this construct in pulp regeneration in-vivo. Methods: DPSCs and HUVECs were encapsulated in PuraMatrix (BD-Biosciences) as mono-cultures or co-cultures at different ratios (3:1, 1:1 1:3). Viability, morphology and three-dimensional organization of cells within PuraMatrix were assessed over 2-weeks under confocal microscopy. Cell-PHS constructs were induced for odonto/osteogenic differentiation (up to 21-days); and examined for alkaline phosphatase (ALP) activity and mineralization (von-Kossa staining). Cell-encapsulated PuraMatrix-constructs were injected into the canal space of full-length tooth-roots and implanted into the subcutaneous space of 6-8-week-old female severe combined immunodeficient mice. Two-to-four weeks after transplantation, mice were euthanized and tooth-roots were removed for histological (Haematoxylin and eosin) and immunohistochemical (human mitochondria, CD31) analysis. Experiments were conducted in triplicate using DPSCs from three different donors and analysed statistically (ANOVA). Results: Results showed that both DPSCs and HUVECs survived well in co-cultures compared to monocultures. HUVECs, when co-cultured with DPSCs formed a vessel-like network throughout the PuraMatrix compared to HUVEC-monocultures where not only HUVECs failed to form any vessels but also underwent apoptosis. ELISA-assay revealed that DPSCs secrete VEGF in high amounts inhibiting apoptosis and promoting vessel formation by HUVECs. Higher ALP activity and mineralization were observed in co-cultures compared to monocultures (p<0.05). Both DPSC-monoculture and co-culture groups showed vascularised pulp-like tissue with patches of osteodentin after transplantation in mice. Co-cultured groups showed higher amounts of extracellular matrix, vascularisation and mineralization compared to DPSC-monocultures in-vivo. Immunohistochemistry for human mitochondria confirmed the contribution of transplanted cells in regenerated pulp-like tissue and vasculature. Conclusion: DPSCs and HUVECs being encapsulated within PuraMatrix demonstrate synergistic effects in odontogenic differentiation and angiogenesis and have potential for engineering vascularised pulp tissues in-vivo.
DescriptionOral Presentation
Session 43: Keynote Address; Dental Pulp Mineralization and Regeneration
Persistent Identifierhttp://hdl.handle.net/10722/199322
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorDissanayaka, WLen_US
dc.contributor.authorHargreaves, Ken_US
dc.contributor.authorJin, Len_US
dc.contributor.authorZhang, Cen_US
dc.date.accessioned2014-07-22T01:13:37Z-
dc.date.available2014-07-22T01:13:37Z-
dc.date.issued2014en_US
dc.identifier.citationThe 92nd General Session & Exhibition of the International Association for Dental Research (IADR), Cape Town, South Africa, 25-28 June 2014. In Journal of Dental Research, 2014, v. 93 n. Special issue B: abstract no. 174en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/199322-
dc.descriptionOral Presentation-
dc.descriptionSession 43: Keynote Address; Dental Pulp Mineralization and Regeneration-
dc.description.abstractObjectives: To co-culture dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs) in an injectable, peptide hydrogel scaffold: PuraMatrix while fabricating a vascular network in-vitro and to utilize this construct in pulp regeneration in-vivo. Methods: DPSCs and HUVECs were encapsulated in PuraMatrix (BD-Biosciences) as mono-cultures or co-cultures at different ratios (3:1, 1:1 1:3). Viability, morphology and three-dimensional organization of cells within PuraMatrix were assessed over 2-weeks under confocal microscopy. Cell-PHS constructs were induced for odonto/osteogenic differentiation (up to 21-days); and examined for alkaline phosphatase (ALP) activity and mineralization (von-Kossa staining). Cell-encapsulated PuraMatrix-constructs were injected into the canal space of full-length tooth-roots and implanted into the subcutaneous space of 6-8-week-old female severe combined immunodeficient mice. Two-to-four weeks after transplantation, mice were euthanized and tooth-roots were removed for histological (Haematoxylin and eosin) and immunohistochemical (human mitochondria, CD31) analysis. Experiments were conducted in triplicate using DPSCs from three different donors and analysed statistically (ANOVA). Results: Results showed that both DPSCs and HUVECs survived well in co-cultures compared to monocultures. HUVECs, when co-cultured with DPSCs formed a vessel-like network throughout the PuraMatrix compared to HUVEC-monocultures where not only HUVECs failed to form any vessels but also underwent apoptosis. ELISA-assay revealed that DPSCs secrete VEGF in high amounts inhibiting apoptosis and promoting vessel formation by HUVECs. Higher ALP activity and mineralization were observed in co-cultures compared to monocultures (p<0.05). Both DPSC-monoculture and co-culture groups showed vascularised pulp-like tissue with patches of osteodentin after transplantation in mice. Co-cultured groups showed higher amounts of extracellular matrix, vascularisation and mineralization compared to DPSC-monocultures in-vivo. Immunohistochemistry for human mitochondria confirmed the contribution of transplanted cells in regenerated pulp-like tissue and vasculature. Conclusion: DPSCs and HUVECs being encapsulated within PuraMatrix demonstrate synergistic effects in odontogenic differentiation and angiogenesis and have potential for engineering vascularised pulp tissues 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.subjectPulp-
dc.subjectRegeneration and Stem Cells-
dc.titlePrevascularized Three-Dimensional DPSC Constructs in Dental Pulp Regenerationen_US
dc.typeConference_Paperen_US
dc.identifier.emailDissanayaka, WL: warunad@hku.hken_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.hkuros231054en_US
dc.identifier.volume93en_US
dc.identifier.issueSpecial issue B: abstract no. 174en_US
dc.publisher.placeUnited States-

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