Department of Fixed Prosthesis & Implant Prosthodontics, School of Dentistry, Aristotle University of Thessaloniki, Greece.
Dent Mater. 2011 Jun;27(6):608-17. doi: 10.1016/j.dental.2011.03.002. Epub 2011 Apr 12.
The aim of this study was to investigate the effects of HEMA and TEGDMA on the odontogenic differentiation potential of dental pulp stem/progenitor cells.
Dental stem/progenitor cell cultures were established from pulp biopsies of human deciduous teeth of 1-3 year-old children (Deciduous Teeth Stem Cells-DTSCs). Cultures were characterized for stem cell markers, including STRO-1, CD146, CD34, CD45 using flow cytometry. Cytotoxicity was evaluated with the MTT assay. DTSCs were then induced for osteo/odontogenic differentiation by media containing dexamethasone, KH(2)PO(4),β-glycerophosphate and L-ascorbic acid phosphate in the presence of nontoxic concentrations of HEMA (0.05-0.5mM) and TEGDMA (0.05-0.25mM) for 3 weeks. Additionally, the effects of a single exposure (72 h) to higher concentrations of HEMA (2mM) and TEGDMA (1mM) were also evaluated.
DTSCs cultures were positive for STRO-1 (7.53±2.5%), CD146 (91.79±5.41%), CD34 (11.87±3.02%) and negative for CD45. In the absence of monomers cell migration, differentiation and production of mineralized dentin-like structures could be observed. Cells also progressively expressed differentiation markers, including dentin sialophosphoprotein-DSPP, bone sialoprotein-BSP, osteocalcin-OCN and alkaline phosphatase-ALP. On the contrary, long-term exposure to nontoxic concentrations of HEMA and TEGDMA significantly delayed the differentiation and mineralization processes of DTSCs, whereas, one time exposure to higher concentrations of these monomers almost completed inhibited mineral nodule formation. BSP, OCN, ALP and DSPP expression were also significantly down-regulated.
These findings suggest that HEMA and TEGDMA can severely disturb the odontogenic differentiation potential of pulp stem/progenitor cells, which might have significant consequences for pulp tissue homeostasis and repair.
本研究旨在探讨 HEMA 和 TEGDMA 对牙髓干细胞/祖细胞成牙分化潜能的影响。
从 1-3 岁儿童乳牙牙髓活检中建立牙髓干细胞/祖细胞培养物（乳牙牙髓干细胞-DTSCs）。使用流式细胞术对 STRO-1、CD146、CD34、CD45 等干细胞标志物进行鉴定。采用 MTT 法评估细胞毒性。然后，在含有地塞米松、KH(2)PO(4)、β-甘油磷酸和 L-抗坏血酸磷酸的培养基中，加入非毒性浓度的 HEMA（0.05-0.5mM）和 TEGDMA（0.05-0.25mM）诱导 DTSCs 向成骨/成牙分化，培养 3 周。此外，还评估了单次暴露于较高浓度 HEMA（2mM）和 TEGDMA（1mM）的 72 小时的影响。
DTSCs 培养物对 STRO-1（7.53±2.5%）、CD146（91.79±5.41%）、CD34（11.87±3.02%）呈阳性，而对 CD45 呈阴性。在没有单体细胞迁移的情况下，可观察到细胞分化和矿化牙本质样结构的产生。细胞还逐渐表达分化标志物，包括牙本质涎磷蛋白-DSPP、骨涎蛋白-BSP、骨钙素-OCN 和碱性磷酸酶-ALP。相反，长期暴露于非毒性浓度的 HEMA 和 TEGDMA 显著延迟了 DTSCs 的分化和矿化过程，而一次暴露于这些单体的较高浓度几乎完全抑制了矿化结节的形成。BSP、OCN、ALP 和 DSPP 的表达也显著下调。
这些发现表明，HEMA 和 TEGDMA 可严重干扰牙髓干细胞/祖细胞的成牙分化潜能，这可能对牙髓组织的稳态和修复产生重大影响。
