IMR Press / FBL / Volume 28 / Issue 5 / DOI: 10.31083/j.fbl2805085
Open Access Original Research
KLF9 Promotes Osteogenic Differentiation of Dental Stem Cells by Negatively Regulating Notch1 Mediated Signaling Pathway
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1 Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University & Department of Endodontic, Affiliated Hospital of Stomatology, Nanjing Medical University, 211166 Nanjing, Jiangsu, China
2 Institute of Stomatology, Nanjing Medical University, 211166 Nanjing, Jiangsu, China
3 Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, 510280 Guangzhou, Guangdong, China
4 Department of Oral and Maxillofacial Surgery, Stomatological Hospital, School of Stomatology, Southern Medical University, 510280 Guangzhou, Guangdong, China
*Correspondence: licui@smu.edu.cn (Li Cui); yujinhua@njmu.edu.cn (Jinhua Yu)
Front. Biosci. (Landmark Ed) 2023, 28(5), 85; https://doi.org/10.31083/j.fbl2805085
Submitted: 5 January 2023 | Revised: 3 February 2023 | Accepted: 17 February 2023 | Published: 6 May 2023
Copyright: © 2023 The Author(s). Published by IMR Press.
This is an open access article under the CC BY 4.0 license.
Abstract

Background: Human dental stem cells (DSCs) are excellent sources of cells for treating dental and craniofacial diseases. However, the mechanisms regulating DSC osteogenic differentiation are still unclear. In this study, we aimed to determine the role of Krüppel-like factor 9 (KLF9) in regulating the biological functions of DSCs and explore the underlying molecular mechanisms. Methods: Bioinformatic analyses, quantitative real-time polymerase chain reaction (qRT‒PCR) and Western blotting were performed to determine the KLF9 level during osteogenic differentiation of DSCs. The effects of KLF9 depletion or overexpression on DSC osteogenic differentiation were then evaluated. The osteogenic potential and associated mineralized nodule-forming activities of DSCs were monitored via Alizarin red S staining and quantitative analyses of osteogenic markers. The regulatory effect of KLF9 on the Notch1 signaling pathway was analyzed by luciferase reporter assays. Results: KLF9 mRNA expression was consistently increased during mesenchymal stem cell osteogenic differentiation in multiple public datasets, and our qRT‒PCR and Western blotting data further validated this finding. In addition, KLF9 depletion promoted proliferation and suppressed osteogenic differentiation of DSCs, while enforced expression of KLF9 promoted the DSC osteogenic potential. Mechanistically, KLF9 negatively regulated the Notch1-mediated signaling pathway by directly binding to the Notch1 promoter. More importantly, Notch1 inhibition/overexpression partially rescued the suppressive/enhancing effects of KLF9 depletion/overexpression on the osteogenic differentiation of DSCs, indicating that Notch1 is a functional downstream target of KLF9. Conclusions: In summary, our results strongly demonstrate that KLF9 is a crucial transcription factor that controls the osteogenic differentiation of DSCs by negatively regulating the Notch1 signaling pathway.

Keywords
dental stem cells
KLF9
osteogenic differentiation
Notch1
signaling pathway
Funding
81873707/National Natural Science Foundation of China
82170940/National Natural Science Foundation of China
81901006/National Natural Science Foundation of China
Figures
Fig. 1.
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