Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.
Clinical Research Center of Shaanxi Province for Dental and Maxillofacial Diseases, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.
Int J Hyperthermia. 2024;41(1):2369749. doi: 10.1080/02656736.2024.2369749. Epub 2024 Jun 26.
Dentin hypersensitivity (DH) is a prevalent condition, but long-term effective treatments are scarce. Differentiation of odontoblast-like cells is promising for inducing tertiary dentinogenesis and ensuring sustained therapeutic efficacy against DH. This study examined the effects and mechanism of action of mild heat stress (MHS) on the differentiation of odontoblast-like MDPC-23 cells.
We used a heating device to accurately control the temperature and duration, mimicking the thermal microenvironment of odontoblast-like cells. Using this device, the effects of MHS on cell viability and differentiation were examined. Cell viability was assessed using the MTT assay. The expression and nucleoplasmic ratio of the yes-associated protein (YAP) were examined by western blotting and immunofluorescence. The gene expression levels of heat shock proteins (HSPs) and dentin matrix protein-1 (DMP1) were measured using qPCR. Dentin sialophosphoprotein (DSPP) expression was evaluated using immunofluorescence and immunoblotting. Verteporfin was used to inhibit YAP activity.
Mild heat stress (MHS) enhanced the odontoblast differentiation of MDPC-23 cells while maintaining cell viability. MHS also increased YAP activity, as well as the levels of HSP25 mRNA, HSP70 mRNA, HSP90α mRNA, DMP1 mRNA, and DSPP protein. However, after YAP inhibition, both cell viability and the levels of HSP90α mRNA, DMP1 mRNA, and DSPP protein were reduced.
YAP plays a crucial role in maintaining cell viability and promoting odontoblast differentiation of MDPC-23 cells under MHS. Consequently, MHS is a potential therapeutic strategy for DH, and boosting YAP activity could be beneficial for maintaining cell viability and promoting odontoblast differentiation.
牙本质敏感（DH）是一种普遍存在的病症，但长期有效的治疗方法却很少。成牙本质细胞样细胞的分化有望诱导第三期牙本质生成，并确保对 DH 的持续治疗效果。本研究旨在探讨温和热应激（MHS）对成牙本质细胞样 MDPC-23 细胞分化的影响及其作用机制。
我们使用加热装置来精确控制温度和时间，模拟成牙本质细胞样细胞的热微环境。利用该装置，研究了 MHS 对细胞活力和分化的影响。通过 MTT 法评估细胞活力。通过 Western blot 和免疫荧光法检测 yes 相关蛋白（YAP）的表达和核质比。通过 qPCR 测量热休克蛋白（HSPs）和牙本质基质蛋白 1（DMP1）的基因表达水平。通过免疫荧光和免疫印迹评估牙本质涎磷蛋白（DSPP）的表达。使用 Verteporfin 抑制 YAP 活性。
温和热应激（MHS）增强了 MDPC-23 细胞的成牙本质分化，同时保持了细胞活力。MHS 还增加了 YAP 活性以及 HSP25mRNA、HSP70mRNA、HSP90αmRNA、DMP1mRNA 和 DSPP 蛋白的水平。然而，在抑制 YAP 后，细胞活力以及 HSP90αmRNA、DMP1mRNA 和 DSPP 蛋白的水平均降低。
YAP 在维持 MHS 下 MDPC-23 细胞活力和促进成牙本质细胞分化中发挥关键作用。因此，MHS 是治疗 DH 的一种潜在治疗策略，增强 YAP 活性可能有助于维持细胞活力和促进成牙本质细胞分化。
