Betaine promotes osteogenic differentiation in immortalized human dental pulp-derived cells

Abstract

Objectives This study aimed to evaluate the effect of betaine (BET) on immortalized human dental pulp stem cell (ihDP) osteogenic differentiation. Materials and methods hDPs were immortalized using SV40 T-antigen transfection. Characterization, multilineage differentiation, proliferation, cell cycle, colony-forming unit, and cellular senescence were evaluated (n = 4). The effect of BET on ihDP response was assessed (n = 4). Osteogenic differentiation was detected using ALP, ARS staining, and RT-qPCR (n = 4). To investigate the involvement of calcium signaling, the cells were pretreated with either 8-(NN-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) or thapsigargin before BET treatment (n = 6). Results ihDPs retained similar phenotypic characteristics presented in hDPs but exhibited an increase in cell proliferation and extended culture to passage 25. An increased proportion of cells in S and G2/M phases without senescence was observed in ihDPs. BET (50 mM) treatment significantly increased mineral deposition at 14 days and upregulated ALP, MSX2, BMP2, and RUNX2 expression. TMB-8 pretreatment reduced the effect of BET-induced ihDP osteogenic differentiation, whereas thapsigargin promoted osteogenic differentiation in ihDPs synergistically with BET. Conclusion ihDPs showed superior proliferation ability and a longer life span, which could serve as a promising cell for regenerative dentistry. BET promoted odonto/osteogenic differentiation via intracellular calcium regulation.

Methylenetetrahydrofolate reductase polymorphisms in dental caries-induced pulp inflammation and regeneration of dentine-pulp complex: Future perspectives

“…To address these future requirements, we hypothesize that MTHFR-SNP-carriers need two-prong approach, which includes i) oral multi-nutrients supplementation (DEN, including betaine) for systemic benefits and ii) chemotaxis induced cell-homing strategy that uses multiple bioactive molecules (betaine, vitamin-D, NAC, vitamin-B12 and methylfolate) to facilitate efficient regeneration of the dentine-pulp complex with new dentine formation, neurogenic and vasculogenic properties. In support of the utilization of the selected bioactive molecules for the proposed regenerative model, vitamins (vitamin-B, -D and -E), NAC and betaine have been reported to enhance the dental regeneration process by eliciting cytoprotective, anti-oxidant and anti-inflammatory effects, ( Escobar et al, 2020 , Fernández-Villa et al, 2018 , Kornsuthisopon et al, 2022 , Meng et al, 2022 , Pei et al, 2018 , Woo et al, 2015 ).…”

Research progress on optimization of in vitro isolation, cultivation and preservation methods of dental pulp stem cells for clinical application

“…The conditioned medium of immortalized DPSCs was collected and further prepared into a pharmaceutical composition for cancer therapy containing 1.5 times more insulin-like growth factor and vascular endothelial cell proliferation factor. Furthermore, Kornsuthisopon et al (2022) immortalized human DPSCs by SV40T antigen transfection, extending their lifespan to the 25th passage with better proliferation ability. The immortalized DPSCs were endowed with enhanced proliferation, migration, and apoptotic abilities without tumorigenicity, and they displayed the same stem cell surface markers as the original DPSCs, according to Li et al (2020) utilization of the iggyBac transposon-mediated method to stably express SV40T-Ag.…”

“…The lifespan of immortalized DPSCs was extended to the 25th passage with better proliferative ability Kornsuthisopon et al (2022) Xiangfen Li SV40 T-antigen The immortalized DPSCs were endowed with enhanced proliferation, migration, and apoptotic abilities without tumorigenicity, and they displayed the same stem cell surface markers as the original DPSCs. The immortalization of DPSCs prepared by this method is reversible, and the cells still maintain high biological characteristics after de-immortalization Li et al (2020) hTERT, human telomerase reverse transcriptase; DPSCs, dental pulp stem cells; IGF, insulin-like growth factor; VEGF, vascular endothelial growth factor.…”