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Conference Paper: cAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells

TitlecAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells
Authors
Issue Date2017
PublisherInternational Association for Dental Research.
Citation
The 31st IADR-SEA & 28th SEAADE Annual Scientific Meeting & 40th CTADS Annual Scientific Meeting, Taipei, Taiwan, 10-13 August 2017, p. S0126 How to Cite?
AbstractObjectives: This study aims to investigate the underlining interplay of cyclic adenosine monophosphate (cAMP) and transforming growth factor-β1 (TGF-β1) on the odonto/osteogenic differentiation of apical papilla-derived stem cells (SCAPs). Methods: SCAPs were isolated from human immature third molars. Forskolin was used to activate and H89 was used to inhibit cAMP signaling in SCAPs. The protein levels of TGF-β1 signaling downstream molecule (Smad3) and phosphorylation of Smad3 (p-Smad3) were assayed by western blot analysis. Smad3 inhibitor (SIS3) was used to inhibit the effect of TGF-β1 signaling on the odonto/osteogenic differentiation in SCAPs. The amount of calcium mineral deposition was detected by Alizarin red staining. Additionally, quantitative real-time polymerase chain reaction (RT-PCR) were taken to evaluate cAMP signaling on TGF-β1-mediated odonto/osteogenic differentiation potential of SCAPs. Results: Protein expression of p-Smad3 decreased in SCAPs stimulated by the Forskolin (Figure 1A); while H89 increased the level of p-Smad3 (Figure 1B). Alizarin red staining showed that TGF-β1 antagonized the up-regulated effect of Forskolin. Blocking TGF-β1 signaling with SIS3 enhanced the mineralized nodule formation of SCAPs (Figure 2). RT-PCR found that in the presence of TGF-β1, SCAPs with Forskolin displayed markedly down-regulation in the expression of odonto/osteogenic marker compared to Forskolin treated only (Forskolin with TGF-β1:0.552±0.243, Forskolin: 1.439±0.105; p<0.004). Besides, the expression of odonto/osteogenic marker of Forskolin with SIS3 and Forskolin were 1.874±0.161 and 1.439±0.105, respectively (p=0.017). Conclusions: This study demonstrated cAMP signaling induced the odonto/osteogenic differentiation of SCAPs via inhibition of TGF-β1-Smad3 signaling. The cAMP signaling pathway could be a strategy target for dentine regeneration in clinical dentistry.
Persistent Identifierhttp://hdl.handle.net/10722/247706

 

DC FieldValueLanguage
dc.contributor.authorZhang, J-
dc.contributor.authorZhang, C-
dc.contributor.authorChu, CH-
dc.date.accessioned2017-10-18T08:31:20Z-
dc.date.available2017-10-18T08:31:20Z-
dc.date.issued2017-
dc.identifier.citationThe 31st IADR-SEA & 28th SEAADE Annual Scientific Meeting & 40th CTADS Annual Scientific Meeting, Taipei, Taiwan, 10-13 August 2017, p. S0126-
dc.identifier.urihttp://hdl.handle.net/10722/247706-
dc.description.abstractObjectives: This study aims to investigate the underlining interplay of cyclic adenosine monophosphate (cAMP) and transforming growth factor-β1 (TGF-β1) on the odonto/osteogenic differentiation of apical papilla-derived stem cells (SCAPs). Methods: SCAPs were isolated from human immature third molars. Forskolin was used to activate and H89 was used to inhibit cAMP signaling in SCAPs. The protein levels of TGF-β1 signaling downstream molecule (Smad3) and phosphorylation of Smad3 (p-Smad3) were assayed by western blot analysis. Smad3 inhibitor (SIS3) was used to inhibit the effect of TGF-β1 signaling on the odonto/osteogenic differentiation in SCAPs. The amount of calcium mineral deposition was detected by Alizarin red staining. Additionally, quantitative real-time polymerase chain reaction (RT-PCR) were taken to evaluate cAMP signaling on TGF-β1-mediated odonto/osteogenic differentiation potential of SCAPs. Results: Protein expression of p-Smad3 decreased in SCAPs stimulated by the Forskolin (Figure 1A); while H89 increased the level of p-Smad3 (Figure 1B). Alizarin red staining showed that TGF-β1 antagonized the up-regulated effect of Forskolin. Blocking TGF-β1 signaling with SIS3 enhanced the mineralized nodule formation of SCAPs (Figure 2). RT-PCR found that in the presence of TGF-β1, SCAPs with Forskolin displayed markedly down-regulation in the expression of odonto/osteogenic marker compared to Forskolin treated only (Forskolin with TGF-β1:0.552±0.243, Forskolin: 1.439±0.105; p<0.004). Besides, the expression of odonto/osteogenic marker of Forskolin with SIS3 and Forskolin were 1.874±0.161 and 1.439±0.105, respectively (p=0.017). Conclusions: This study demonstrated cAMP signaling induced the odonto/osteogenic differentiation of SCAPs via inhibition of TGF-β1-Smad3 signaling. The cAMP signaling pathway could be a strategy target for dentine regeneration in clinical dentistry.-
dc.languageeng-
dc.publisherInternational Association for Dental Research. -
dc.relation.ispartofInternational Association for Dental Research South East Asian Division Meeting (IADR-SEA)-
dc.titlecAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells-
dc.typeConference_Paper-
dc.identifier.emailZhang, J: jing817@hku.hk-
dc.identifier.emailZhang, C: zhangcf@hku.hk-
dc.identifier.emailChu, CH: chchu@hku.hk-
dc.identifier.authorityZhang, C=rp01408-
dc.identifier.authorityChu, CH=rp00022-
dc.identifier.hkuros280887-
dc.identifier.spageS0126-
dc.identifier.epageS0126-
dc.publisher.placeTaipei, Taiwan-

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