Department of Prosthodontics, School & Hospital of Stomatology, Wenzhou Medical University, Wenzhou, China.
Department of Nursing, School & Hospital of Stomatology, Wenzhou Medical University, Wenzhou, China.
Bioengineered. 2022 Apr;13(4):11201-11213. doi: 10.1080/21655979.2022.2063495.
p75NTR, a neural crest stem cell marker, is continuously expressed in mesenchymal cells during tooth development. Importantly, high expression of p75NTR in the late bell stage implicates its involvement in odontogenesis and mineralization. However, the regulatory mechanisms underlying p75NTR involvement in odonto/osteogenic differentiation remain unclear. Here, we investigate the effect and potential mechanisms underlying p75NTR involvement in odonto/osteogenic differentiation. We dissected EMSCs from the first branchial arches of mice embryo and compared the proliferation and migration of p75NTR and p75NTREMSCs by transwell, scratch and cell counting kit 8(CCK8)assays. The differentiation ability and the involvement of nuclear factor kappa-B (NF-κB) pathway were investigated through alkaline phosphatase and immunofluorescence assay, real-time PCR, and western blot. During induction of dental epithelium conditioned medium, p75NTR EMSCs exhibited deeper Alkaline phosphatase (ALP) staining and higher expression of odonto/osteogenic genes/proteins (e.g., dentin sialoprotein (DSPP) than p75NTR EMSCs. Moreover, p75NTR EMSCs exhibited higher nuclear P65 expression than p75NTREMSCs. Inhibition of NF-κB pathway with Bay11-7082 in p75NTREMSCs substantially decreased DSPP expression level. However, activation of NF-κB pathway with Bay11-7082 in p75NTREMSCs enhanced DSPP expression level. Thus, p75NTR possibly plays a paramount role in the proliferation and differentiation of EMSCs via NF-κB pathway.
p75NTR 是一种神经嵴干细胞标志物，在牙齿发育过程中持续在间充质细胞中表达。重要的是，在钟状晚期 p75NTR 的高表达暗示其参与了牙发生和矿化。然而，p75NTR 参与牙/骨向分化的调节机制尚不清楚。在这里，我们研究了 p75NTR 参与牙/骨向分化的作用和潜在机制。我们从小鼠胚胎第一鳃弓中分离出 EMSCs，并通过 Transwell、划痕和 CCK8 检测比较了 p75NTR 和 p75NTR+EMSCs 的增殖和迁移。通过碱性磷酸酶和免疫荧光染色、实时 PCR 和 Western blot 检测了分化能力和核因子 kappa-B (NF-κB) 途径的参与。在牙上皮条件培养基诱导过程中，p75NTR+EMSCs 表现出更深的碱性磷酸酶（ALP）染色和更高的牙/骨向基因/蛋白表达（例如，牙本质涎磷蛋白（DSPP））。此外，p75NTR+EMSCs 中核 P65 的表达高于 p75NTR-EMSCs。用 Bay11-7082 抑制 NF-κB 途径可显著降低 p75NTR-EMSCs 中 DSPP 的表达水平。然而，用 Bay11-7082 激活 p75NTR-EMSCs 中的 NF-κB 途径可增强 DSPP 的表达水平。因此，p75NTR 可能通过 NF-κB 途径在 EMSCs 的增殖和分化中发挥重要作用。
