Platelet-rich plasma driven quiescence–activation reprogramming primes dental pulp stem cells for translational bone regeneration

Dental pulp stem cells (DPSCs) are multipotent cells widely used in tissue repair and regeneration, making effective preconditioning strategies essential to maximize their therapeutic potential. Here, we aimed to reactivate quiescent DPSCs using platelet-rich plasma (PRP) and evaluate its effects on proliferation, multilineage differentiation, and senescence, leveraging quiescence–activation dynamics to enhance their regenerative efficacy. DPSCs were cultured in serum-free basal medium to induce a quiescent state. Subsequently, fetal bovine serum (FBS), human platelet lysate (HPL), PRP, and a commercial serum‑free supplement for human DPSCs (S-DPSC) were added to parallel cultures to reactivate the quiescent cells. Activated DPSCs were then assessed for their proliferative capacity, senescence resistance, and multilineage differentiation potential. PRP served as an effective activator, whereas HPL demonstrated relatively limited activation capacity. Notably, Re+PRP significantly promoted DPSCs proliferation without inducing excessive replicative senescence. Transcriptomic analysis indicated that different activators trigger distinct signaling pathways. Moreover, assessment of osteogenic differentiation-related gene expression, alkaline phosphatase activity, and Alizarin Red S staining consistently demonstrated significantly enhanced osteogenic potential in the Re+PRP compared with the other groups. In contrast, chondrogenic differentiation potential did not differ significantly between Re+PRP and +FBS. The quiescence–activation protocol using PRP significantly enhances DPSCs proliferative capacity and osteogenic differentiation potential. Clinical significance: This study presents a preconditioning strategy for DPSCs that aims to enhance their therapeutic potential and facilitate their clinical translation for bone regeneration. • A quiescence–activation preconditioning protocol was developed for DPSCs. • PRP possesses the functional property to reactivate quiescent DPSCs, a capacity that HPL lacks. • PRP reactivates quiescent DPSCs through the synergistic effects of blood components. • PRP reactivation of quiescent DPSCs potentiates proliferative capacity and osteogenic differentiation.