TY - JOUR AB - Stem cell‑based therapy may provide a novel approach for neural tissue regeneration. A small molecule cocktail‑based culture protocol was previously shown to enhance neurogenic differentiation of stem cells from dental tissues. The present study aimed to investigate the early phase of small molecule‑induced neurogenic differentiation of stem cells from the apical papilla (SCAP). SCAP were cultured in neural‑induction medium or neural‑induction medium with small molecules (NIMS‑SCAP) and examined for their cell morphologies. Expression levels of neural progenitor cell‑related markers, including Nestin, paired‑box gene 6 (Pax6) and Sry‑related HMG box 2 (Sox2), were examined using western blotting and immunocytofluorescence. Expression of differentiated neuron‑related markers, including neurofilament protein (NFM), neuron‑specific nuclear protein (NeuN) and microtubule‑associated protein (MAP)‑2, were also examined using western blotting, while NFM and MAP2 gene expression and cell proliferation were assessed using reverse transcription‑quantitative (RT‑q)PCR and Cell Counting Kit (CCK)‑8 assays, respectively. SCAP morphology was affected by small molecules after as little as 30 min. Specifically, Nestin, Pax6 and Sox2 expression detected using western blotting was increased by day 3 but then decreased over the course of 7 days with neural induction, while immunocytofluorescence revealed expression of all three markers in NIMS‑SCAP. The protein levels of NFM, NeuN and MAP2 on day 7 were significantly upregulated in NIMS‑SCAP, as detected using western blotting, while NFM and MAP2 gene expression levels detected using RT‑qPCR were significantly increased on days 5 and 7. Proliferation of NIMS‑SCAP ceased after 5 days. Electrophysiological analysis showed that only SCAP cultured in NIMS had the functional activity of neuronal cells. Thus, small molecules reprogrammed SCAP into neural progenitor cells within the first 3 days, followed by further differentiation into neuron‑like cells. AD - Restorative Dental Sciences, Endodontology, Faculty of Dentistry, The University of Hong Kong, Hong Kong 999077, SAR, P.R. China Department of Implant Dentistry, The Affiliated Stomatological Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221002, P.R. China School of Stomatology, Peking University, Beijing 100081, P.R. China Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China AU - Chen,Qixin AU - Yuan,Changyong AU - Jiang,Shan AU - Heng,Boon Chin AU - Zou,Ting AU - Shen,Zhongshan AU - Wang,Penglai AU - Zhang,Chengfei DA - 2021/06/01 DO - 10.3892/etm.2021.9978 IS - 6 JO - Exp Ther Med KW - small molecule stem cells apical papilla differentiation neurogenesis PY - 2021 SN - 1792-0981 1792-1015 SP - 546 ST - Small molecules efficiently reprogram apical papilla stem cells into neuron‑like cells T2 - Experimental and Therapeutic Medicine TI - Small molecules efficiently reprogram apical papilla stem cells into neuron‑like cells UR - https://doi.org/10.3892/etm.2021.9978 VL - 21 ER -