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Conference Paper: ECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Model
Title | ECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Model |
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Authors | |
Keywords | Dentin Endodontics Pulp Regeneration and Tissue engineering |
Issue Date | 2012 |
Publisher | Sage 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? |
Abstract | Objectives: 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. |
Description | Session: Pulp Biology and Regeneration |
Persistent Identifier | http://hdl.handle.net/10722/182062 |
ISSN | 2023 Impact Factor: 5.7 2023 SCImago Journal Rankings: 1.909 |
DC Field | Value | Language |
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dc.contributor.author | Dissanayaka, WL | en_US |
dc.contributor.author | Zhang, C | en_US |
dc.contributor.author | Jin, L | en_US |
dc.contributor.author | Hargreaves, KM | en_US |
dc.date.accessioned | 2013-04-17T07:20:43Z | - |
dc.date.available | 2013-04-17T07:20:43Z | - |
dc.date.issued | 2012 | en_US |
dc.identifier.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 | en_US |
dc.identifier.issn | 0022-0345 | - |
dc.identifier.uri | http://hdl.handle.net/10722/182062 | - |
dc.description | Session: Pulp Biology and Regeneration | - |
dc.description.abstract | Objectives: 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.language | eng | en_US |
dc.publisher | Sage Publications, Inc.. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925 | - |
dc.relation.ispartof | Journal of Dental Research | en_US |
dc.rights | Journal of Dental Research. Copyright © Sage Publications, Inc.. | - |
dc.subject | Dentin | - |
dc.subject | Endodontics | - |
dc.subject | Pulp | - |
dc.subject | Regeneration and Tissue engineering | - |
dc.title | ECs Enhance DPSC Properties And Therapeutic Potential In Three-dimensional Model | en_US |
dc.type | Conference_Paper | en_US |
dc.identifier.email | Zhang, C: zhangcf@hku.hk | en_US |
dc.identifier.email | Jin, L: ljjin@hkucc.hku.hk | en_US |
dc.identifier.authority | Zhang, C=rp01408 | en_US |
dc.identifier.authority | Jin, L=rp00028 | en_US |
dc.identifier.hkuros | 213910 | en_US |
dc.identifier.volume | 91 | en_US |
dc.identifier.issue | Special Issue C: abstract no. 168954 | en_US |
dc.publisher.place | United States | - |
dc.identifier.issnl | 0022-0345 | - |