Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1

Transforming growth factor-β1 (TGF-β1) is an important multifunctional cytokine with dual effects on stem cell differentiation. However, the role of TGF-β1 on odontogenic differentiation of dental pulp stem cells (DPSCs) remains to be entirely elucidated. In the present study, we initially investiga...

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Main Authors: Yu Bai, Xin Liu, Junqing Li, Zhihua Wang, Qian Guo, Min Xiao, Paul R. Cooper, Qing Yu, Wenxi He
Format: Article
Language:English
Published: Wiley 2022-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2022/2361376
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author Yu Bai
Xin Liu
Junqing Li
Zhihua Wang
Qian Guo
Min Xiao
Paul R. Cooper
Qing Yu
Wenxi He
author_facet Yu Bai
Xin Liu
Junqing Li
Zhihua Wang
Qian Guo
Min Xiao
Paul R. Cooper
Qing Yu
Wenxi He
author_sort Yu Bai
collection DOAJ
description Transforming growth factor-β1 (TGF-β1) is an important multifunctional cytokine with dual effects on stem cell differentiation. However, the role of TGF-β1 on odontogenic differentiation of dental pulp stem cells (DPSCs) remains to be entirely elucidated. In the present study, we initially investigated the effect of TGF-β1 at a range of concentrations (0.1-5 ng/mL) on the proliferation, cell cycle, and apoptosis of DPSCs. Subsequently, to determine the effect of TGF-β1 on odontogenic differentiation, alkaline phosphatase (ALP) activity and Alizarin Red S (ARS) staining assays at different concentrations and time points were performed. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis were used to determine the levels of odonto-/osteo-genic differentiation-related gene and protein expression, respectively. For in vivo studies, newly formed tissue was assessed by Masson’s trichrome and von Kossa staining. Data indicated that TGF-β1 inhibited DPSCs proliferation in a concentration-and time-dependent manner (p<0.05) and induced cell cycle arrest but did not affect apoptosis. ALP activity was enhanced, while ARS reduced gradually with increasing TGF-β1 concentrations, accompanied by increased expression of early marker genes of odonto-/osteo-genic differentiation and decreased expression of late-stage mineralization marker genes (p<0.05). ALP expression was elevated in the TGF-β1-treatment group until 14 days, and the intensity of ARS staining was attenuated at days 14 and 21 (p<0.05). Compared with the control group, abundant collagen but no mineralized tissues were observed in the TGF-β1-treatment group in vivo. Overall, these findings indicate that TGF-β1 promotes odontogenic differentiation of DPSCs at early-stage while inhibiting later-stage mineralization processes.
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spelling doaj-art-7f4a69b7fc2f4a23b3fed79d523d9bcc2025-08-20T03:35:40ZengWileyStem Cells International1687-96782022-01-01202210.1155/2022/2361376Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1Yu Bai0Xin Liu1Junqing Li2Zhihua Wang3Qian Guo4Min Xiao5Paul R. Cooper6Qing Yu7Wenxi He8State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyDepartment of Oral SciencesState Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of StomatologyDepartment of StomatologyTransforming growth factor-β1 (TGF-β1) is an important multifunctional cytokine with dual effects on stem cell differentiation. However, the role of TGF-β1 on odontogenic differentiation of dental pulp stem cells (DPSCs) remains to be entirely elucidated. In the present study, we initially investigated the effect of TGF-β1 at a range of concentrations (0.1-5 ng/mL) on the proliferation, cell cycle, and apoptosis of DPSCs. Subsequently, to determine the effect of TGF-β1 on odontogenic differentiation, alkaline phosphatase (ALP) activity and Alizarin Red S (ARS) staining assays at different concentrations and time points were performed. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis were used to determine the levels of odonto-/osteo-genic differentiation-related gene and protein expression, respectively. For in vivo studies, newly formed tissue was assessed by Masson’s trichrome and von Kossa staining. Data indicated that TGF-β1 inhibited DPSCs proliferation in a concentration-and time-dependent manner (p<0.05) and induced cell cycle arrest but did not affect apoptosis. ALP activity was enhanced, while ARS reduced gradually with increasing TGF-β1 concentrations, accompanied by increased expression of early marker genes of odonto-/osteo-genic differentiation and decreased expression of late-stage mineralization marker genes (p<0.05). ALP expression was elevated in the TGF-β1-treatment group until 14 days, and the intensity of ARS staining was attenuated at days 14 and 21 (p<0.05). Compared with the control group, abundant collagen but no mineralized tissues were observed in the TGF-β1-treatment group in vivo. Overall, these findings indicate that TGF-β1 promotes odontogenic differentiation of DPSCs at early-stage while inhibiting later-stage mineralization processes.http://dx.doi.org/10.1155/2022/2361376
spellingShingle Yu Bai
Xin Liu
Junqing Li
Zhihua Wang
Qian Guo
Min Xiao
Paul R. Cooper
Qing Yu
Wenxi He
Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
Stem Cells International
title Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
title_full Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
title_fullStr Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
title_full_unstemmed Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
title_short Stage-Dependent Regulation of Dental Pulp Stem Cell Odontogenic Differentiation by Transforming Growth Factor-β1
title_sort stage dependent regulation of dental pulp stem cell odontogenic differentiation by transforming growth factor β1
url http://dx.doi.org/10.1155/2022/2361376
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