Exploring the neurogenic differentiation of human dental pulp stem cells

• Published in: PloS one 2022-11, Vol.17 (11), p.e0277134-e0277134
• Main Authors: Al-Maswary, Arwa A, O'Reilly, Molly, Holmes, Andrew P, Walmsley, A. Damien, Cooper, Paul R, Scheven, Ben A
• Format: Article
• Language: English
• Subjects: Acetylcholinesterase; Acetyltransferase; Analysis; Biology and Life Sciences; Brain-derived neurotrophic factor; Brn-3 protein; Cell culture; Cell differentiation; Choline; Choline O-acetyltransferase; Cholinergics; Dental pulp; Dentistry; Differentiation (biology); Endodontics; Extracellular signal-regulated kinase; Health aspects; Homeobox; Injuries; Kinases; MAP kinase; Medical examination; Medicine and Health Sciences; Microscopy; Nervous system; Nestin; Neural stem cells; Neuroblastoma; Neurosciences; Parkinson's disease; Parkinsons disease; Peripherin; Phosphopyruvate hydratase; Phosphorylation; Physiology; Potassium channels (voltage-gated); Potassium currents; Protein kinase; Proteins; Recovery of function; Regeneration; Retinoic acid; Services; Social Sciences; Sodium; Sodium channels (voltage-gated); Spinal cord; Stem cell transplantation; Stem cells; Synapsin I; Transplantation; Tubulin
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0277134

Human dental pulp stem cells (hDPSCs) have increasingly gained interest as a potential therapy for nerve regeneration in medicine and dentistry, however their neurogenic potential remains a matter of debate. This study aimed to characterize hDPSC neuronal differentiation in comparison with the human SH-SY5Y neuronal stem cell differentiation model. Both hDPSCs and SH-SY5Y could be differentiated to generate typical neuronal-like cells following sequential treatment with all-trans retinoic acid (ATRA) and brain-derived neurotrophic factor (BDNF), as evidenced by significant expression of neuronal proteins [beta]III-tubulin (TUBB3) and neurofilament medium (NF-M). Both cell types also expressed multiple neural gene markers including growth-associated protein 43 (GAP43), enolase 2/neuron-specific enolase (ENO2/NSE), synapsin I (SYN1), nestin (NES), and peripherin (PRPH), and exhibited measurable voltage-activated Na.sup.+ and K.sup.+ currents. In hDPSCs, upregulation of acetylcholinesterase (ACHE), choline O-acetyltransferase (CHAT), sodium channel alpha subunit 9 (SCN9A), POU class 4 homeobox 1 (POU4F1/BRN3A) along with a downregulation of motor neuron and pancreas homeobox 1 (MNX1) indicated that differentiation was more guided toward a cholinergic sensory neuronal lineage. Furthermore, the Extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor U0126 significantly impaired hDPSC neuronal differentiation and was associated with reduction of the ERK1/2 phosphorylation. In conclusion, this study demonstrates that extracellular signal-regulated kinase/Mitogen-activated protein kinase (ERK/MAPK) is necessary for sensory cholinergic neuronal differentiation of hDPSCs. hDPSC-derived cholinergic sensory neuronal-like cells represent a novel model and potential source for neuronal regeneration therapies.