The Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland.
Stem Cells Dev. 2019 Aug 1;28(15):1050-1058. doi: 10.1089/scd.2019.0023. Epub 2019 Jul 15.
The aim of this study was to compare the in vitro osteogenic differentiation potential of within-subject mesenchymal stem cells (MSCs) derived from the dental pulp of permanent teeth (dental pulp stem cells-DPSCs), the dental pulp of deciduous teeth (stem cells from human exfoliated deciduous teeth-SHEDs), and the periodontal ligament of permanent teeth (periodontal ligament stem cells-PDLSCs). A single subject was identified that required concurrent removal of both deciduous and permanent teeth for orthodontic purposes. Primary, mixed population cells from dental pulp, deciduous dental pulp, and periodontal ligament were obtained by the tissue outgrowth method. Subsequently, isolation of STRO-1 +ve cells from their respective primary cell cultures was achieved by immunomagnetic separation. Cells were induced with an osteogenic cocktail of 5 mM β-glycerophosphate, 100 nM dexamethasone, and 50 mg/mL ascorbic acid for up to 21 days. Osteogenic responses were assessed functionally by an alkaline phosphatase (ALP) activity assay and an alizarin red staining assay. Expression of the early osteogenic associated genes, alkaline phosphatase gene (), runt-related transcription factor 2 (), collagen type I alpha 1 (), and secreted phosphoprotein 1 (), was compared by qPCR at days 1, 4, and 7 of differentiation. Functional analysis revealed that there were significant differences in intracellular ALP activity on days 4, 7, 10, and 14 with PDLSCs > SHEDs > DPSCs. Quantification of alizarin red staining showed significantly more mineralization for PDLSCs by day 21. Gene expression analysis showed significant early upregulations of the osteogenic markers and for PDLSCs over DPSCs and SHEDs. SHEDs showed significantly higher upregulation of over DPSCs. In conclusion, PDLSCs showed a significantly higher osteogenic differentiation potential than both DPSCs and SHEDs evidenced by functional studies and gene expression. This may be of significance for the use of dentally derived MSCs in bone tissue engineering applications.
本研究旨在比较来源于恒牙牙髓（牙髓干细胞-DPSCs）、乳牙牙髓（人脱落乳牙牙髓干细胞-SHEDs）和恒牙牙周膜（牙周膜干细胞-PDLSCs）的同一体内间充质干细胞（MSCs）的体外成骨分化潜能。确定了一位因正畸需要同时拔除乳牙和恒牙的患者。通过组织外生法获得牙髓、乳牙牙髓和牙周膜的原代混合细胞群体。随后，通过免疫磁珠分离从各自的原代细胞培养物中分离出 STRO-1+ve 细胞。用成骨细胞鸡尾酒（5mM β-甘油磷酸、100nM 地塞米松和 50mg/mL 抗坏血酸）诱导细胞培养物，培养时间最长达 21 天。通过碱性磷酸酶（ALP）活性测定和茜素红染色测定来评估成骨反应。通过 qPCR 在分化的第 1、4 和 7 天比较早期成骨相关基因碱性磷酸酶基因（）、 runt 相关转录因子 2（）、Ⅰ型胶原α 1 （）和分泌型磷蛋白 1（）的表达。功能分析显示，在第 4、7、10 和 14 天细胞内 ALP 活性存在显著差异，PDLSCs＞SHEDs＞DPSCs。茜素红染色定量显示，PDLSCs 在第 21 天的矿化程度明显更高。基因表达分析显示，在 PDLSCs 中，成骨标志物和在早期的上调显著高于 DPSCs 和 SHEDs。SHEDs 中 基因的上调显著高于 DPSCs。综上所述，PDLSCs 在功能研究和基因表达方面均显示出比 DPSCs 和 SHEDs 更高的成骨分化潜能，这对于 dentally 衍生的 MSCs 在骨组织工程应用中的应用具有重要意义。
