Escobar Lina M, Bendahan Zita, Bayona Andrea, Castellanos Jaime E, González María-Clara
Unidad de Manejo Integral de Malformaciones Craneofaciales (UMIMC), Facultad de Odontología, Universidad El Bosque, Bogotá, Colombia.
Posgrado de Ortodoncia, Facultad de Odontología, Universidad El Bosque, Bogotá, Colombia.
Int J Dent. 2020 Jul 1;2020:8860840. doi: 10.1155/2020/8860840. eCollection 2020.
The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs).
In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR.
Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes.
Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.
本研究的目的是确定维生素D和维生素E对人牙髓干细胞（hDPSCs）增殖、形态和分化的影响。
在这项体外实验研究中，分离并鉴定了hDPSCs，然后分别或联合使用不同剂量和治疗周期的维生素D和维生素E对其进行处理。分别使用光学显微镜和刃天青测定法评估形态变化和细胞增殖。使用茜素红S染色以及实时逆转录聚合酶链反应（RT-PCR）检测RUNX2、Osterix和骨钙素基因的表达来评估成骨细胞分化。
与未处理的细胞相比，在144小时后，用维生素D（49%）、维生素E（35%）和维生素D+E（61%）处理后细胞数量显著减少。与单独使用维生素处理的细胞培养物相比，用维生素D+E处理的细胞汇合度降低，细胞质更广泛且更扁平，在处理7天后开始形成大量钙化结节。14天后，用维生素D、维生素E和维生素D+E处理增加了RUNX2、Osterix和骨钙素基因的转录。
维生素D和维生素E诱导hDPSCs向成骨细胞分化，这表现为细胞增殖减少、形态变化以及钙化结节的形成，同时分化基因的表达增加。维生素D+E联合处理在向成骨细胞谱系的分化中诱导协同效应。
