Department of Restorative Dentistry and Endodontics, Faculty of Dental Medicine, Lebanese University, Beirut, Lebanon.
Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon.
J Dent. 2020 Oct;101:103413. doi: 10.1016/j.jdent.2020.103413. Epub 2020 Jun 22.
Dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) are types of human dental tissue-derived mesenchymal stem cells (MSCs) that have emerged as an interesting and promising source of stem cells in the field of tissue engineering. The aim of this work is to isolate stem cells from DPSCs and SHED, cultivate them in vitro and compare their odontogenic differentiation potential.
DPSCs and SHED were extracted from molars, premolars and canines of six healthy subjects aged 5-29 years. The cells were characterized, using flow cytometry, for mesenchymal stem cell surface markers. MTT colorimetric assay was applied to assess cell viability. Alizarin red staining, alkaline phosphatase (ALP) activity, quantitative real-time PCR (qRT-PCR) and western blot were carried out to determine DPSCs and SHED osteogenic/odontogenic differentiation.
DPSCs express higher STRO-1 and CD44 levels compared to SHED. Moreover, the cells differentiate and acquire columnar shape with a level of calcium deposition and mineralization that is the same between DPSCs and SHED. ALP activity, ALP, COLI, DMP-1, DSPP, OC, and RUNX2 (osteogenic/odontogenic differentiation markers) expression levels were higher in DPSCs.
DPSCs and SHED express MSCs markers. Although both cell types had calcium deposits, DPSCs presented a higher ALP activity level. In addition, DPSCs showed higher levels of osteogenic and odontogenic differentiation markers such as COLI, DSPP, OC, RUNX2, and DMP-1. These results suggest that DPSCs are closer to the phenotype of odontoblasts than SHED and may improve the efficacy of human dental tissue-derived mesenchymal stem cells therapeutic protocols.
'CLINICAL SIGNIFICANCE': DPSCs are closer than t SHED to the phenotype of odontoblasts. This would be helpful to enable better therapeutic decisions when applying MSCs-based therapy in the field of dentistry.
牙髓干细胞（DPSCs）和人乳牙脱落干细胞（SHED）是人类牙齿组织来源的间充质干细胞（MSCs）的两种类型，它们在组织工程领域成为一种有趣且有前途的干细胞来源。本研究的目的是分离 DPSCs 和 SHED 中的干细胞，在体外培养并比较其成牙分化潜能。
从 6 名 5-29 岁健康受试者的磨牙、前磨牙和尖牙中提取 DPSCs 和 SHED。采用流式细胞术对间充质干细胞表面标志物进行鉴定。MTT 比色法评估细胞活力。茜素红染色、碱性磷酸酶（ALP）活性、实时定量 PCR（qRT-PCR）和 Western blot 用于鉴定 DPSCs 和 SHED 的成骨/成牙分化。
与 SHED 相比，DPSCs 表达更高水平的 STRO-1 和 CD44。此外，两种细胞均分化为具有柱状形态，且具有相同水平的钙沉积和矿化。ALP 活性、ALP、COLI、DMP-1、DSPP、OC 和 RUNX2（成骨/成牙分化标志物）的表达水平在 DPSCs 中更高。
DPSCs 和 SHED 表达 MSC 标志物。尽管两种细胞类型均有钙沉积，但 DPSCs 具有更高的 ALP 活性水平。此外，DPSCs 表达更高水平的成骨和成牙分化标志物，如 COL1、DSPP、OC、RUNX2 和 DMP-1。这些结果表明，DPSCs 比 SHED 更接近成牙本质细胞的表型，并且可能提高人类牙齿组织来源的间充质干细胞治疗方案的疗效。
DPSCs 比 SHED 更接近成牙本质细胞的表型。这将有助于在牙科领域应用基于 MSC 的治疗时做出更好的治疗决策。
