Clock genes are expressed in cementum and regulate the proliferation and mineralization of cementoblasts.

Gufeng Liu, Quan Sun, Xiaoyi Wu, Ying Liu, Yang Chen, Zhengguo Cao, Yanhe Hu, Haibin Xia

In vitro cellular & developmental biology. Animal 2023 Jan

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Circadian clock genes are present in the ameloblasts, odontoblasts, and dental pulp cells. The cementum plays a vital role in connecting the roots of teeth to the alveolar bone by anchoring the periodontal ligament. The present study aimed at confirming the existence of clock genes and describing the potential regulatory effects of REV-ERBα in the cementum. The tooth-periodontal ligament-alveolar bone complexes of 6-week-old mice were analyzed using immunohistochemistry. OCCM-30 cells, an immortalized cementoblast cell line, were synchronized with dexamethasone. We used RT-PCR to detect the expression of clock genes in the absence or presence of SR8278, an effective antagonist of REV-ERBα. We performed a cell counting kit-8 (CCK-8) assay to determine the effect of SR8278 on cell proliferation. RT-PCR and Western blot were used to measure the expression of mineralization-related markers in mineralization-induced OCCM-30 cells, with or without SR8278 treatment. Finally, we used Alizarin red staining, and ALP staining and activity to further verify the effect of SR8278 on mineralization of OCCM-30 cells on macro-level. In our study, clock protein expression was confirmed in the murine cementum. Clock genes were shown to oscillate continuously in OCCM-30 cells. SR8278-induced inactivation of REV-ERBα inhibited the proliferation but promoted the mineralization of OCCM-30 cells. The present study confirmed the presence of clock genes in the cementum, where they potentially participate in cell proliferation and mineralization. Our findings may inspire new research directions for periodontal regeneration via clock gene manipulation. © 2023. The Society for In Vitro Biology.

Citation

Gufeng Liu, Quan Sun, Xiaoyi Wu, Ying Liu, Yang Chen, Zhengguo Cao, Yanhe Hu, Haibin Xia. Clock genes are expressed in cementum and regulate the proliferation and mineralization of cementoblasts. In vitro cellular & developmental biology. Animal. 2023 Jan;59(1):76-84