Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis
Abstract Background Orthodontic tooth movement inevitably induces cementum resorption, which is an urgent problem for orthodontists to confront. Human periodontal ligament stem cells (hPDLSCs) exert an important role in the orthodontic tooth movement and exhibit multidirectional differentiation abil...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-09-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-03149-8 |
| _version_ | 1798002980161585152 |
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| author | Zuping Wu Yuying He Sirui Chen Li Zhu Jiahe Wang Demao Zhang Jing Xie Shujuan Zou Chenchen Zhou |
| author_facet | Zuping Wu Yuying He Sirui Chen Li Zhu Jiahe Wang Demao Zhang Jing Xie Shujuan Zou Chenchen Zhou |
| author_sort | Zuping Wu |
| collection | DOAJ |
| description | Abstract Background Orthodontic tooth movement inevitably induces cementum resorption, which is an urgent problem for orthodontists to confront. Human periodontal ligament stem cells (hPDLSCs) exert an important role in the orthodontic tooth movement and exhibit multidirectional differentiation ability in cementum regeneration. Connective tissue growth factor (CTGF) is an important extracellular matrix protein for bone homeostasis and cell differentiation. The purpose of our study was to explore the role of CTGF in cementum repair and cementogenesis and to elucidate its underlying mechanism. Methods A cementum defect model was established by tooth movement with heavy forces, and the cementum repair effect of CTGF was observed via micro-CT, HE staining and immunohistochemical staining. RT‒qPCR, western blotting (WB), alizarin red staining and ALP activity experiments verified the mineralization ability of hPDLSCs stimulated with CTGF. The expression of Cx43 in periodontal ligament cells was detected by WB and immunofluorescence (IF) experiments after CTGF stimulation in vivo and in vitro. Subsequently, the mineralization ability of hPDLSCs was observed after application of CTGF and the small interfering RNA Si-Cx43. Additionally, co-intervention via application of the small interfering RNA Si-CTGF and the Cx43 agonist ATRA in hPDLSCs was performed to deepen the mechanistic study. Next, WB, IF experiments and co-immunoprecipitation were conducted to confirm whether CTGF triggers the Cx43/β-catenin axis to regulate cementoblast differentiation of hPDLSCs. Results Local oral administration of CTGF to the cementum defects in vivo facilitated cementum repair. CTGF facilitated the cementogenesis of hPDLSCs in a concentration-dependent manner. Cx43 acted as a downstream effector of CTGF to regulate cementoblast differentiation. Si-Cx43 reduced CTGF-induced cementoblast differentiation. The Cx43 agonist ATRA restored the low differentiation capacity induced by Si-CTGF. Further mechanistic studies showed that CTGF triggered the activation of β-catenin in a dose-dependent manner. In addition, co-localization IF analysis and co-immunoprecipitation demonstrated that Cx43 interacted with β-catenin at cell‒cell connections. Si-Cx43 attenuated the substantial expression of β-catenin induced by CTGF. The Cx43 agonist reversed the inhibition of β-catenin induced by Si-CTGF. IF demonstrated that the nuclear importation of β-catenin was related to the immense expression of Cx43 at cell‒cell junctions. Conclusions Taken together, these data demonstrate that CTGF promotes cementum repair and cementogenesis through activation of the Cx43/β-catenin signalling axis. |
| first_indexed | 2024-04-11T12:00:58Z |
| format | Article |
| id | doaj.art-835b7542dc6b473d99d7ede8d8d14ef3 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-11T12:00:58Z |
| publishDate | 2022-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-835b7542dc6b473d99d7ede8d8d14ef32022-12-22T04:24:51ZengBMCStem Cell Research & Therapy1757-65122022-09-0113111810.1186/s13287-022-03149-8Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axisZuping Wu0Yuying He1Sirui Chen2Li Zhu3Jiahe Wang4Demao Zhang5Jing Xie6Shujuan Zou7Chenchen Zhou8State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityState Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan UniversityAbstract Background Orthodontic tooth movement inevitably induces cementum resorption, which is an urgent problem for orthodontists to confront. Human periodontal ligament stem cells (hPDLSCs) exert an important role in the orthodontic tooth movement and exhibit multidirectional differentiation ability in cementum regeneration. Connective tissue growth factor (CTGF) is an important extracellular matrix protein for bone homeostasis and cell differentiation. The purpose of our study was to explore the role of CTGF in cementum repair and cementogenesis and to elucidate its underlying mechanism. Methods A cementum defect model was established by tooth movement with heavy forces, and the cementum repair effect of CTGF was observed via micro-CT, HE staining and immunohistochemical staining. RT‒qPCR, western blotting (WB), alizarin red staining and ALP activity experiments verified the mineralization ability of hPDLSCs stimulated with CTGF. The expression of Cx43 in periodontal ligament cells was detected by WB and immunofluorescence (IF) experiments after CTGF stimulation in vivo and in vitro. Subsequently, the mineralization ability of hPDLSCs was observed after application of CTGF and the small interfering RNA Si-Cx43. Additionally, co-intervention via application of the small interfering RNA Si-CTGF and the Cx43 agonist ATRA in hPDLSCs was performed to deepen the mechanistic study. Next, WB, IF experiments and co-immunoprecipitation were conducted to confirm whether CTGF triggers the Cx43/β-catenin axis to regulate cementoblast differentiation of hPDLSCs. Results Local oral administration of CTGF to the cementum defects in vivo facilitated cementum repair. CTGF facilitated the cementogenesis of hPDLSCs in a concentration-dependent manner. Cx43 acted as a downstream effector of CTGF to regulate cementoblast differentiation. Si-Cx43 reduced CTGF-induced cementoblast differentiation. The Cx43 agonist ATRA restored the low differentiation capacity induced by Si-CTGF. Further mechanistic studies showed that CTGF triggered the activation of β-catenin in a dose-dependent manner. In addition, co-localization IF analysis and co-immunoprecipitation demonstrated that Cx43 interacted with β-catenin at cell‒cell connections. Si-Cx43 attenuated the substantial expression of β-catenin induced by CTGF. The Cx43 agonist reversed the inhibition of β-catenin induced by Si-CTGF. IF demonstrated that the nuclear importation of β-catenin was related to the immense expression of Cx43 at cell‒cell junctions. Conclusions Taken together, these data demonstrate that CTGF promotes cementum repair and cementogenesis through activation of the Cx43/β-catenin signalling axis.https://doi.org/10.1186/s13287-022-03149-8CTGFCx43Cementumβ-CateninHuman periodontal stem cells |
| spellingShingle | Zuping Wu Yuying He Sirui Chen Li Zhu Jiahe Wang Demao Zhang Jing Xie Shujuan Zou Chenchen Zhou Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis Stem Cell Research & Therapy CTGF Cx43 Cementum β-Catenin Human periodontal stem cells |
| title | Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis |
| title_full | Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis |
| title_fullStr | Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis |
| title_full_unstemmed | Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis |
| title_short | Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis |
| title_sort | connective tissue growth factor promotes cementogenesis and cementum repair via cx43 β catenin axis |
| topic | CTGF Cx43 Cementum β-Catenin Human periodontal stem cells |
| url | https://doi.org/10.1186/s13287-022-03149-8 |
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