Transcription factor EB enhances odontoblastic differentiation in dental pulp stem cells.

Transcription factor EB induces glycolysis and mitophagy in dental pulp stem cells.

Glycolysis and mitophagy form a positive feedback loop attenuates differentiation.

To evaluate the regulatory effect of transcription factor EB (TFEB) on the odontoblastic differentiation of dental pulp stem cells(DPSCs) in vivo and in vitro.

RNA-seq was used to detect differentially expressed genes in differentiated DPSCs. Lysosomes and the expression of the related gene TFEB were examined in DPSCs. DPSCs were then transfected with lentivirus for TFEB-overexpression. Cell proliferation was detected using CCK-8 and EdU assays, while cell differentiation was detected using ALP and ARS detection kits. Subsequently, mitophagy and cell metabolism were examined using TEM and Seahorse. An odontoblastic differentiation model was constructed subcutaneously in nude mice. Finally, the effects of glycolysis and mitophagy inhibitors were evaluated on odontoblastic differentiation and the associated mechanisms were explored.

TFEB overexpression promoted a significant increase in ALP activity and the expression of differentiation-related genes in DPSCs, while it inhibited cell proliferation. In vivo, TFEB overexpression caused higher bone volume/trabecular volume(BV/TV), and an increase in collagen formation and heightened DMP-1 expression. Furthermore, Seahorse flux analysis demonstrated that TFEB promoted metabolic reprogramming. Transmission electron microscope(TEM) results indicated an increase in mitochondrial autophagosomes after TFEB overexpression, and the expression of mitophagy-related genes was also elevated. The odontoblastic differentiation of DPSCs promoted by TFEB overexpression was suppressed after the addition of 2-DG and Midiv-1. Addition of Midiv-1 reduced the glycolytic rate of DPSCs, while addition of 2-DG also decreased the mitophagy level of the cells.

Our results showed that TFEB promoted the odontoblastic differentiation of DPSCs and identified mitophagy and metabolic reprogramming as a positive feedback loop.

Human dental pulp stem cells

Kyoto Encyclopedia of Genes and Genomes

Quantitative real-time polymerase chain reaction

Runt-related transcription factor 2

Dentinsialophosphoprotein

Dentin matrix acidic phosphoprotein 1

transcription factor EB

microtubule- associated proteins 1 A and 1B

Mitochondrial division inhibitor 1

PTEN induced putative kinase 1

Transcription factor EB

Dental pulp stem cells

Mitophagy

Glycolysis

Odontoblast

© 2024 The Authors. Published by Elsevier Ltd.