Investigations into the basal neural-like properties of dental pulp stem cells reveals they possess a functional type 2 muscarinic receptor which regulates quiescence
Dental pulp stem cells (DPSCs) are a population of mesenchyme-derived cells residing within the dental pulp known for their multipotent differentiation potential and neural-like properties. While a functional cholinergic system has been described in various mesenchymal stem cell (MSC) populations, its presence and role in DPSCs remain unexplored. This study investigated the neurotransmitter-associated gene expression profile of human DPSCs using a targeted array panel and immunocytochemistry. The functionality of acetylcholine receptors (AChRs) was assessed through pharmacological activation and inhibition using receptor-specific agonists and antagonists. Cellular responses to cholinergic modulation, including effects on viability and proliferation, were evaluated via LDH release, CCK-8, and annexin V/PI staining assays. Among the 92 neurotransmitter-associated genes profiled, 43 were expressed, including multiple muscarinic receptors and nicotinic receptor subunits. Protein level expression of the m2, m3, and m5 muscarinic receptors was confirmed. Furthermore, activation of the m2AChR, using arecaidine propargyl ester hydrobromide (APE), led to a reversible reduction in DPSC proliferation without compromising cell viability. The findings demonstrate that DPSCs exhibit cholinoceptive properties and identify the m2AChR as a key modulator of cell cycle arrest. This receptor may serve as a therapeutic target for regulating DPSC behaviour and maintaining stem cell quiescence in regenerative applications. Overall design: RNA-Seq profiling of untreated DPSCs and DPSCs stiumulatied with 100 µM arecaidine propargyl ester hydrobromide (APE) for 4 and 24 hours.