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Conference Paper: Synergistic effects of PDL stem cells/HUVECs in 3D cell sheet

TitleSynergistic effects of PDL stem cells/HUVECs in 3D cell sheet
Authors
KeywordsCell biology
Periodontal disease
Periodontics
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 91st General Session & Exhibition of the International Association for Dental Research (IADR), Seattle, Washington, USA, 20-23 March 2013. In Journal of Dental Research, 2013, v. 92 n. Special Issue A: abstract no. 3900 How to Cite?
AbstractObjective: To investigate the expression of odontogenic differentiation markers and vascular network formation in 3-D cell sheet with different cell ratios of periodontal ligament stem cells (PDLSCs) and human umbilical vein endothelial cells (HUVECs). Method: Human PDLSCs were isolated and characterized by flow-cytometry, and HUVECs were used for construction of cell sheet. Both types of cells were seeded on temperature responsive culture dishes with PDLSCs alone, HUVECs alone, and various ratios of these cells (1:1, 2:1, 5:1 and 1:5) to obtain confluent cell sheet layers. The expression of odontogenic pathway markers including ALP, BSP, RUNX2 were analyzed at 3 and 7 days using RT-PCR. Further ALP protein quantification was done at 7 and 14days using alkaline phosphatase assay. The calcium nodule formation was assessed qualitatively and quantitatively by Alizarin Red assay. Histological evaluations of three cell sheet constructs treated with different combinations (PDLSCs-PDLSCs-PDLSCs/PDLSCs-HUVECs-PDLSCs/Co-Culture 2:1 PDLSCs-HUVECs) were performed with H&E and immunofluorescence straining. Statistical analysis was done by t-test(p<0.05). Result: Significantly higher ALP gene expression was observed at 3days in 1:1 (PDLSCs:HUVECs) (2.52±0.67) and 5:1(4.05±1.07) co-culture groups compared with other groups(p<0.05), being consistent with ALP protein quantification. However, BSP and RUNX2 genes expressions were higher at 7days compared to that detected at 3 days. Significant calcium mineralization was detected and quantified by Alizarin red assay at 14days in 1:1(1323.55±6.54µm) and 5:1(994.67±4.15µm) co-cultures as compared with mono-culture cell sheets(p<0.05). H&E and CD31 immuno-staining illustrated the development of a layered cell sheet structure with endothelial cell islands within the constructed PDLSCs-HUVECs-PDLSCs and co-culture groups. Furthermore, HUVECs permeated into layered cell sheet, was suggestive of rudimentary vascular network initiation. Conclusion: This study suggests that PDLSCs-HUVECs co-culture models exhibit significantly high levels of odontogenic markers with signs of initial vascualar formation. This novel 3-D cell sheet-based approach may be potentially beneficial for periodontal regenerative therapy. This abstract is based on research that was funded entirely or partially by an outside source: Seed Funding Programme for Basic Research, The University of Hong Kong. 201111159191
DescriptionPoster Presentation
Session 425: Differentiation of Periodontal Cells
Persistent Identifierhttp://hdl.handle.net/10722/183225
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorPanduwawala, KCPPen_US
dc.contributor.authorSamaranayake, LPen_US
dc.contributor.authorJin, Len_US
dc.contributor.authorZhang, Cen_US
dc.date.accessioned2013-05-15T01:48:20Z-
dc.date.available2013-05-15T01:48:20Z-
dc.date.issued2013en_US
dc.identifier.citationThe 91st General Session & Exhibition of the International Association for Dental Research (IADR), Seattle, Washington, USA, 20-23 March 2013. In Journal of Dental Research, 2013, v. 92 n. Special Issue A: abstract no. 3900en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/183225-
dc.descriptionPoster Presentation-
dc.descriptionSession 425: Differentiation of Periodontal Cells-
dc.description.abstractObjective: To investigate the expression of odontogenic differentiation markers and vascular network formation in 3-D cell sheet with different cell ratios of periodontal ligament stem cells (PDLSCs) and human umbilical vein endothelial cells (HUVECs). Method: Human PDLSCs were isolated and characterized by flow-cytometry, and HUVECs were used for construction of cell sheet. Both types of cells were seeded on temperature responsive culture dishes with PDLSCs alone, HUVECs alone, and various ratios of these cells (1:1, 2:1, 5:1 and 1:5) to obtain confluent cell sheet layers. The expression of odontogenic pathway markers including ALP, BSP, RUNX2 were analyzed at 3 and 7 days using RT-PCR. Further ALP protein quantification was done at 7 and 14days using alkaline phosphatase assay. The calcium nodule formation was assessed qualitatively and quantitatively by Alizarin Red assay. Histological evaluations of three cell sheet constructs treated with different combinations (PDLSCs-PDLSCs-PDLSCs/PDLSCs-HUVECs-PDLSCs/Co-Culture 2:1 PDLSCs-HUVECs) were performed with H&E and immunofluorescence straining. Statistical analysis was done by t-test(p<0.05). Result: Significantly higher ALP gene expression was observed at 3days in 1:1 (PDLSCs:HUVECs) (2.52±0.67) and 5:1(4.05±1.07) co-culture groups compared with other groups(p<0.05), being consistent with ALP protein quantification. However, BSP and RUNX2 genes expressions were higher at 7days compared to that detected at 3 days. Significant calcium mineralization was detected and quantified by Alizarin red assay at 14days in 1:1(1323.55±6.54µm) and 5:1(994.67±4.15µm) co-cultures as compared with mono-culture cell sheets(p<0.05). H&E and CD31 immuno-staining illustrated the development of a layered cell sheet structure with endothelial cell islands within the constructed PDLSCs-HUVECs-PDLSCs and co-culture groups. Furthermore, HUVECs permeated into layered cell sheet, was suggestive of rudimentary vascular network initiation. Conclusion: This study suggests that PDLSCs-HUVECs co-culture models exhibit significantly high levels of odontogenic markers with signs of initial vascualar formation. This novel 3-D cell sheet-based approach may be potentially beneficial for periodontal regenerative therapy. This abstract is based on research that was funded entirely or partially by an outside source: Seed Funding Programme for Basic Research, The University of Hong Kong. 201111159191-
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.subjectCell biology-
dc.subjectPeriodontal disease-
dc.subjectPeriodontics-
dc.subjectRegeneration and Tissue engineering-
dc.titleSynergistic effects of PDL stem cells/HUVECs in 3D cell sheeten_US
dc.typeConference_Paperen_US
dc.identifier.emailSamaranayake, LP: lakshman@hku.hken_US
dc.identifier.emailJin, L: ljjin@hkucc.hku.hken_US
dc.identifier.emailZhang, C: zhangcf@hku.hken_US
dc.identifier.authoritySamaranayake, LP=rp00023en_US
dc.identifier.authorityJin, L=rp00028en_US
dc.identifier.authorityZhang, C=rp01408en_US
dc.identifier.hkuros214404en_US
dc.identifier.volume92en_US
dc.identifier.issueSpecial Issue A: abstract no. 3900en_US
dc.publisher.placeUnited States-

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