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Conference Paper: HUVECs Synergize Osteo/Odontogenic Differentiation Of PDLSCs In 3D Cell Sheets

TitleHUVECs Synergize Osteo/Odontogenic Differentiation Of PDLSCs In 3D Cell Sheets
Authors
KeywordsPeriodontal disease
Periodontics
Periodontium-gingiva
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. 42 How to Cite?
AbstractObjective: To investigate the expression of osteo/odontogenic differentiation markers and vascular network formation in a 3-D cell sheet with varying cell ratios of periodontal ligament stem cells (PDLSCs) and human umbilical vein endothelial cells (HUVECs). Method: Human PDLSCs were isolated and characterized and co-cultured with HUVECs. 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). The expressions of osteo/odontogenic pathway markers ALP, BSP and RUNX2 were analyzed at 3 and 7 days using RT-PCR. Further, ALP protein quantification was performed at 7 and 14 days using alkaline phosphatase assay. The calcium nodule formation was assessed qualitatively and quantitatively by alizarin red assay. Histological evaluations of cell sheet constructs with combinations (PDLSCs-PDLSCs-PDLSCs/PDLSCs-HUVECs-PDLSCs/Co-Culture-Co-culture-Co-culture) were observed with scanning electron microscope (SEM), H&E and immunofluorescence staining. Cell viability was assessed with the live/dead assay. Statistical analysis was performed using 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, the expression of BSP and RUNX2 genes was higher at 7days compared to 3days. Significant calcium mineralization was detected as 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). SEM, H&E and CD31 immuno-staining exemplified the development of a layered cell sheet structure with endothelial cell islands within the constructed PDLSCs-HUVECs-PDLSCs and co-culture groups. HUVECs invaded the layered cell sheet, suggestive of rudimentary vascular network initiation. After 7 days, majority of the cells were viable in all 3 groups. Conclusion: PDLSCs-HUVECs co-culture cell sheet model exhibits significantly high levels of osteo/odontogenic markers with signs of initial vascular 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: This work is supported by seed funding programme for Basic Research,The University of Hong Kong. 201111159191
DescriptionConference Theme: We are the Future
Oral Presentation
Session 7: O7
Persistent Identifierhttp://hdl.handle.net/10722/192570
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorPanduwawala, CPen_US
dc.contributor.authorSamaranayake, LPen_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. 42en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/192570-
dc.descriptionConference Theme: We are the Future-
dc.descriptionOral Presentation-
dc.descriptionSession 7: O7-
dc.description.abstractObjective: To investigate the expression of osteo/odontogenic differentiation markers and vascular network formation in a 3-D cell sheet with varying cell ratios of periodontal ligament stem cells (PDLSCs) and human umbilical vein endothelial cells (HUVECs). Method: Human PDLSCs were isolated and characterized and co-cultured with HUVECs. 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). The expressions of osteo/odontogenic pathway markers ALP, BSP and RUNX2 were analyzed at 3 and 7 days using RT-PCR. Further, ALP protein quantification was performed at 7 and 14 days using alkaline phosphatase assay. The calcium nodule formation was assessed qualitatively and quantitatively by alizarin red assay. Histological evaluations of cell sheet constructs with combinations (PDLSCs-PDLSCs-PDLSCs/PDLSCs-HUVECs-PDLSCs/Co-Culture-Co-culture-Co-culture) were observed with scanning electron microscope (SEM), H&E and immunofluorescence staining. Cell viability was assessed with the live/dead assay. Statistical analysis was performed using 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, the expression of BSP and RUNX2 genes was higher at 7days compared to 3days. Significant calcium mineralization was detected as 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). SEM, H&E and CD31 immuno-staining exemplified the development of a layered cell sheet structure with endothelial cell islands within the constructed PDLSCs-HUVECs-PDLSCs and co-culture groups. HUVECs invaded the layered cell sheet, suggestive of rudimentary vascular network initiation. After 7 days, majority of the cells were viable in all 3 groups. Conclusion: PDLSCs-HUVECs co-culture cell sheet model exhibits significantly high levels of osteo/odontogenic markers with signs of initial vascular 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: This work is supported by 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.subjectPeriodontal disease-
dc.subjectPeriodontics-
dc.subjectPeriodontium-gingiva-
dc.subjectRegeneration and Tissue engineering-
dc.titleHUVECs Synergize Osteo/Odontogenic Differentiation Of PDLSCs In 3D Cell Sheetsen_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.hkuros226800en_US
dc.identifier.volume92en_US
dc.identifier.issueSpecial Issue B: abstract no. 42en_US
dc.publisher.placeUnited States-

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