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Conference Paper: cAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells
Title | cAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells |
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Authors | |
Issue Date | 2017 |
Publisher | International 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, presentation no. S0126 How to Cite? |
Abstract | Objectives: 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. |
Description | Poster Session 1 Salivary / Pulp/ Stemcell Biology - Final Presentation ID: 0126 |
Persistent Identifier | http://hdl.handle.net/10722/247706 |
DC Field | Value | Language |
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dc.contributor.author | Zhang, J | - |
dc.contributor.author | Zhang, C | - |
dc.contributor.author | Chu, CH | - |
dc.date.accessioned | 2017-10-18T08:31:20Z | - |
dc.date.available | 2017-10-18T08:31:20Z | - |
dc.date.issued | 2017 | - |
dc.identifier.citation | The 31st IADR-SEA & 28th SEAADE Annual Scientific Meeting & 40th CTADS Annual Scientific Meeting, Taipei, Taiwan, 10-13 August 2017, presentation no. S0126 | - |
dc.identifier.uri | http://hdl.handle.net/10722/247706 | - |
dc.description | Poster Session 1 Salivary / Pulp/ Stemcell Biology - Final Presentation ID: 0126 | - |
dc.description.abstract | Objectives: 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.language | eng | - |
dc.publisher | International Association for Dental Research. | - |
dc.relation.ispartof | International Association for Dental Research South East Asian Division Meeting (IADR-SEA) | - |
dc.title | cAMP signaling up-regulates odonto/osteogenic differentiation of apical papilla-derived stem cells | - |
dc.type | Conference_Paper | - |
dc.identifier.email | Zhang, J: jing817@hku.hk | - |
dc.identifier.email | Zhang, C: zhangcf@hku.hk | - |
dc.identifier.email | Chu, CH: chchu@hku.hk | - |
dc.identifier.authority | Zhang, C=rp01408 | - |
dc.identifier.authority | Chu, CH=rp00022 | - |
dc.identifier.hkuros | 280887 | - |
dc.identifier.hkuros | 329490 | - |
dc.identifier.spage | S0126 | - |
dc.identifier.epage | S0126 | - |
dc.publisher.place | Taipei, Taiwan | - |