3D culture inhibits replicative senescence of SCAPs via UQCRC2-mediated mitochondrial oxidative phosphorylation.

Stem cells derived from the apical papilla (SCAPs) play a crucial role in tooth root development and dental pulp regeneration. They are important seed cells for bone/tooth tissue engineering. However, replicative senescence remains an unavoidable issue as in vitro amplification increases. This study investigated the effect of a three-dimensional (3D) culture environment constructed with methylcellulose on SCAPs senescence. It was observed that 3D culture conditions can delay cellular senescence, potentially due to changes in mitochondrial function and oxidative phosphorylation. Transcriptome high-throughput sequencing technology revealed that the different mitochondrial states may be related to UQCRC2. Knocking down UQCRC2 expression in the 3D culture group resulted in increased production of mitochondrial reactive oxygen species, decreased mitochondrial membrane potential, and a decline in the oxygen consumption rate for oxidative phosphorylation, accelerating cell senescence. The results of this study indicated that 3D culture can mitigate SCAPs aging by maintaining UQCRC2-mediated mitochondrial homeostasis. These findings provide a new solution for the senescence of SCAPs during in vitro amplification and can promote the applications of SCAPs-based clinical translation.