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Article: In Vitro Analysis of Scaffold-free Prevascularized Microtissue Spheroids Containing Human Dental Pulp Cells and Endothelial Cells
Title | In Vitro Analysis of Scaffold-free Prevascularized Microtissue Spheroids Containing Human Dental Pulp Cells and Endothelial Cells |
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Authors | |
Keywords | scaffold Angiogenesis tissue engineering prevascularization regeneration |
Issue Date | 2015 |
Citation | Journal of Endodontics, 2015, v. 41, n. 5, p. 663-670 How to Cite? |
Abstract | © 2015 American Association of Endodontists.Introduction: Scaffolds often fail to mimic essential functions of the physiologic extracellular matrix (ECM) that regulates cell-cell communication in tissue microenvironments. The development of scaffold-free microtissues containing stem cell-derived ECM may serve as a successful alternative to the use of artificial scaffolds. The current study aimed to fabricate 3-dimensional microtissue spheroids of dental pulp cells (DPCs) prevascularized by human umbilical vein endothelial cells (HUVECs) and to characterize these scaffold-free spheroids for the in vitro formation of pulplike tissue constructs. Methods: Three-dimensional microtissue spheroids of DPC alone and DPC-HUVEC co-cultures were fabricated using agarose micro-molds. Cellular organization within the spheroids and cell viability (live/dead assay) were assessed at days 1, 7, and 14. Microtissue spheroids were allowed to self-assemble into macrotissues, induced for odontogenic differentiation (21 days), and examined for expression levels of osteo/odontogenic markers: alkaline phosphatase, bone sialoprotein and RUNX2 (Real-time PCR), mineralization (von-Kossa), and prevascularisation (immunohistochemistry for CD31). Results: The DPC microtissue microenvironment supported HUVEC survival and capillary network formation in the absence of a scaffolding material and external angiogenic stimulation. Immunohistochemical staining for CD31 showed the capillary network formed by HUVECs did sustain - for a prolonged period - even after the microtissues transformed into a macrotissue. Induced, prevascularized macrotissues showed enhanced differentiation capacity compared with DPC alone macrotissues, as shown by higher osteo/odontogenic gene expression levels and mineralization. Conclusions: These findings provide insight into the complex intercellular cross talk occurring between DPCs and HUVECs in the context of angiogenesis and pulp regeneration and highlight the significance of developing a favorable 3-dimensional microenvironment that can, in turn, contribute toward successful pulp regeneration strategies. |
Persistent Identifier | http://hdl.handle.net/10722/236242 |
ISSN | 2023 Impact Factor: 3.5 2023 SCImago Journal Rankings: 1.356 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Dissanayaka, Waruna Lakmal | - |
dc.contributor.author | Zhu, Lifang | - |
dc.contributor.author | Hargreaves, Kenneth M. | - |
dc.contributor.author | Jin, Lijian | - |
dc.contributor.author | Zhang, Chengfei | - |
dc.date.accessioned | 2016-11-11T07:43:19Z | - |
dc.date.available | 2016-11-11T07:43:19Z | - |
dc.date.issued | 2015 | - |
dc.identifier.citation | Journal of Endodontics, 2015, v. 41, n. 5, p. 663-670 | - |
dc.identifier.issn | 0099-2399 | - |
dc.identifier.uri | http://hdl.handle.net/10722/236242 | - |
dc.description.abstract | © 2015 American Association of Endodontists.Introduction: Scaffolds often fail to mimic essential functions of the physiologic extracellular matrix (ECM) that regulates cell-cell communication in tissue microenvironments. The development of scaffold-free microtissues containing stem cell-derived ECM may serve as a successful alternative to the use of artificial scaffolds. The current study aimed to fabricate 3-dimensional microtissue spheroids of dental pulp cells (DPCs) prevascularized by human umbilical vein endothelial cells (HUVECs) and to characterize these scaffold-free spheroids for the in vitro formation of pulplike tissue constructs. Methods: Three-dimensional microtissue spheroids of DPC alone and DPC-HUVEC co-cultures were fabricated using agarose micro-molds. Cellular organization within the spheroids and cell viability (live/dead assay) were assessed at days 1, 7, and 14. Microtissue spheroids were allowed to self-assemble into macrotissues, induced for odontogenic differentiation (21 days), and examined for expression levels of osteo/odontogenic markers: alkaline phosphatase, bone sialoprotein and RUNX2 (Real-time PCR), mineralization (von-Kossa), and prevascularisation (immunohistochemistry for CD31). Results: The DPC microtissue microenvironment supported HUVEC survival and capillary network formation in the absence of a scaffolding material and external angiogenic stimulation. Immunohistochemical staining for CD31 showed the capillary network formed by HUVECs did sustain - for a prolonged period - even after the microtissues transformed into a macrotissue. Induced, prevascularized macrotissues showed enhanced differentiation capacity compared with DPC alone macrotissues, as shown by higher osteo/odontogenic gene expression levels and mineralization. Conclusions: These findings provide insight into the complex intercellular cross talk occurring between DPCs and HUVECs in the context of angiogenesis and pulp regeneration and highlight the significance of developing a favorable 3-dimensional microenvironment that can, in turn, contribute toward successful pulp regeneration strategies. | - |
dc.language | eng | - |
dc.relation.ispartof | Journal of Endodontics | - |
dc.subject | scaffold | - |
dc.subject | Angiogenesis | - |
dc.subject | tissue engineering | - |
dc.subject | prevascularization | - |
dc.subject | regeneration | - |
dc.title | In Vitro Analysis of Scaffold-free Prevascularized Microtissue Spheroids Containing Human Dental Pulp Cells and Endothelial Cells | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.joen.2014.12.017 | - |
dc.identifier.scopus | eid_2-s2.0-84929030543 | - |
dc.identifier.hkuros | 244440 | - |
dc.identifier.volume | 41 | - |
dc.identifier.issue | 5 | - |
dc.identifier.spage | 663 | - |
dc.identifier.epage | 670 | - |
dc.identifier.isi | WOS:000354597200015 | - |
dc.identifier.issnl | 0099-2399 | - |