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Updated: Jun 6, 2026
Isolation, Culture, and Characterization of Dental Pulp Stem Cells from Human Deciduous and Permanent Teeth
Published on: May 17, 2024
Extracellular vesicles derived from stem cells of human exfoliated deciduous teeth alleviate pulpitis through
Qing Mu1,2, Hui Lu1,2, Pei Li1,2
1Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, 510055, Guangdong, China.
Background:
Pulpitis is an inflammatory condition of the dental pulp, in which excessive immune responses and mitochondrial dysfunction are implicated. Extracellular vesicles (EVs) derived from stem cells of human exfoliated deciduous teeth (SHED) have shown potential in modulating inflammation, yet the underlying mechanisms remain unclear. This study aimed to investigate whether SHED-EVs attenuate mitochondrial dysfunction in dental pulp stem cells (DPSCs) and thereby alleviate pulpitis.
Methods:
SHED-EVs were isolated by ultracentrifugation and characterized. Inflammatory DPSCs (iDPSCs) were induced by lipopolysaccharide (LPS) treatment. The effects of SHED-EVs on proliferation and migration of DPSCs and iDPSCs were assessed using CCK-8, EdU, wound healing, and transwell assays. Inflammation, apoptosis, and mitochondrial function were evaluated by qRT-PCR, ELISA, Western blot, flow cytometry, and mitochondrial assays. RNA sequencing and bioinformatics analyses were performed to identify key regulatory pathways. In vivo, a rat pulpitis model was established, and the therapeutic efficacy of SHED-EVs was assessed by histological and immunohistochemical analyses.
Results:
SHED-EVs were efficiently internalized by DPSCs and enhanced their proliferation and migration capacities. In iDPSCs, SHED-EVs significantly suppressed the expression of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, and alleviated cell apoptosis with reduction of cleaved caspase-3 levels and upregulation of Bcl-2. Notably, SHED-EVs decreased mitochondrial reactive oxygen species (mtROS), improved mitochondrial membrane potential (MMP), and increased mitochondrial length. Transcriptomic and Western blot analyses revealed that SHED-EVs modulated inflammatory signaling pathways by inhibiting NF-κB p65 phosphorylation, downregulating NLRP3, and enhancing FoxO1 expression. Furthermore, SHED-EVs alleviated inflammatory infiltration, with reduced IL-1β and IL-6 expression and restored FoxO1 activity in inflamed rat pulp tissue.
Conclusions:
SHED-EVs alleviate pulpitis by mitigating mitochondrial dysfunction, potentially through modulation of the NF-κB and FoxO1 pathways. These findings suggest that SHED-EVs have significant anti-inflammatory potential and may represent promising candidates for pulpitis treatment.
