PER2 regulates odontoblastic differentiation of dental papilla cells in vitro via intracellular ATP content and reactive oxygen species levels

Haozhen Ma; Xinyue Sheng; Wanting Chen; Hongwen He; Jiawei Liu; Yifan He; Fang Huang

doi:10.7717/peerj.16489

PER2 regulates odontoblastic differentiation of dental papilla cells in vitro via intracellular ATP content and reactive oxygen species levels

PeerJ. 2023 Dec 7:11:e16489. doi: 10.7717/peerj.16489. eCollection 2023.

Authors

Haozhen Ma^{1

2

3}, Xinyue Sheng^{1

2

3}, Wanting Chen^{1

2

3}, Hongwen He^{1

2

3}, Jiawei Liu^{1

2

3}, Yifan He^{1

2

3}, Fang Huang^{1

2

3}

Affiliations

¹ Hospital of Stomatology, Sun Yat-sen University, Guangzhou, China.
² Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, China.
³ Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, China.

Abstract

Background: Dental papilla cells (DPCs) are one of the key stem cells for tooth development, eventually forming dentin and pulp. Previous studies have reported that PER2 is expressed in a 24-hour oscillatory pattern in DPCs in vitro. In vivo, PER2 is highly expressed in odontoblasts (which are differentiated from DPCs). However, whether PER2 modulates the odontogenic differentiation of DPCs is uncertain. This research was to identify the function of PER2 in the odontogenic differentiation of DPCs and preliminarily explore its mechanisms.

Methods: We monitored the expression of PER2 in DPCs differentiated in vivo. We used PER2 overexpression and knockdown studies to assess the role of PER2 in DPC differentiation and performed intracellular ATP content and reactive oxygen species (ROS) assays to further investigate the mechanism.

Results: PER2 expression was considerably elevated throughout the odontoblastic differentiation of DPCs in vivo. Overexpressing Per2 boosted levels of odontogenic differentiation markers, such as dentin sialophosphoprotein (Dspp), dentin matrix protein 1 (Dmp1), and alkaline phosphatase (Alp), and enhanced mineralized nodule formation in DPCs. Conversely, the downregulation of Per2 inhibited the differentiation of DPCs. Additionally, downregulating Per2 further affected intracellular ATP content and ROS levels during DPC differentiation.

Conclusion: Overall, we demonstrated that PER2 positively regulates the odontogenic differentiation of DPCs, and the mechanism may be related to mitochondrial function as shown by intracellular ATP content and ROS levels.

Keywords: ATP; Dental papilla cells; In vitro; Odontoblastic differentiation; PER2; ROS; Tooth development.

MeSH terms

Adenosine Triphosphate
Cell Differentiation / genetics
Dental Papilla*
Odontoblasts*
Reactive Oxygen Species

Substances

Reactive Oxygen Species
Adenosine Triphosphate

Grants and funding

This work was supported by the China Postdoctoral Science Foundation (Grant No. 2022M723593), the Guangdong Basic and Applied Basic Research Foundation (Grant No. 2022A1515110434), the National Natural Science Foundation of China No. 81870737, the Natural Science Foundation of Guangdong Province No. 2021A1515011779 and the Guangdong Financial Fund for High-Caliber Hospital Construction No. 174-2018-XMZC-0001-03-0125/D-02. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.