Hospital of Stomatology, Zunyi Medical University, Zunyi, 563000, Guizhou, China.
School of Stomatology, Southern Medical University, Guangzhou, 510000, Guangdong, China.
Sci Rep. 2024 Aug 21;14(1):19354. doi: 10.1038/s41598-024-70123-0.
Fibroblast growth factor 2 (FGF2) is a crucial factor in odontoblast differentiation and dentin matrix deposition, which facilitates pulpodentin repair and regeneration. Nevertheless, the specific biological function of FGF2 in odontoblastic differentiation remains unclear because it is controlled by complex signalling pathways. This study aimed to investigate the mechanism underlying the effect of FGF2 on osteo/odontogenic differentiation of stem cells from the apical papilla (SCAP). SCAP were pretreated with conditioned media containing FGF2 for 1 week, followed by culturing in induced differentiation medium for another week. RNA sequencing (RNA-seq) combined with quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to evaluate the pathways affected by FGF2 in SCAP. Osteo/odontogenic differentiation of SCAP was determined using Alizarin red S staining, alkaline phosphatase staining, RT-qPCR, and western blotting. Pretreatment with FGF2 for 1 week increased the osteo/odontogenic differentiation ability of SCAP. RNA-seq and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that phosphatidylinositol 3-kinase (PI3K)/AKT signalling is involved in the osteogenic function of FGF2. RT-qPCR results indicated that SCAP expressed FGF receptors, and western blotting showed that p-AKT was reduced in FGF2-pretreated SCAP. The activation of the PI3K/AKT pathway partially reversed the stimulatory effect of FGF2 on osteo/odontogenic differentiation of SCAP. Our findings suggest that pretreatment with FGF2 enhances the osteo/odontogenic differentiation ability of SCAP by inhibiting the PI3K/AKT pathway.
成纤维细胞生长因子 2（FGF2）是牙本质细胞分化和牙本质基质沉积的关键因子，有助于牙髓牙本质修复和再生。然而，FGF2 在牙本质细胞分化中的具体生物学功能尚不清楚，因为它受复杂的信号通路控制。本研究旨在探讨 FGF2 对根尖乳头干细胞（SCAP）成骨/成牙本质分化的作用机制。SCAP 用含有 FGF2 的条件培养基预处理 1 周，然后在诱导分化培养基中培养 1 周。RNA 测序（RNA-seq）结合定量逆转录聚合酶链反应（RT-qPCR）用于评估 FGF2 对 SCAP 中受影响的途径。通过茜素红 S 染色、碱性磷酸酶染色、RT-qPCR 和 Western blot 测定 SCAP 的成骨/成牙本质分化。FGF2 预处理 1 周可提高 SCAP 的成骨/成牙本质分化能力。RNA-seq 和京都基因与基因组百科全书通路分析表明，磷脂酰肌醇 3-激酶（PI3K）/AKT 信号通路参与 FGF2 的成骨功能。RT-qPCR 结果表明，SCAP 表达 FGF 受体，Western blot 显示 FGF2 预处理的 SCAP 中 p-AKT 减少。PI3K/AKT 通路的激活部分逆转了 FGF2 对 SCAP 成骨/成牙本质分化的刺激作用。我们的研究结果表明，FGF2 预处理通过抑制 PI3K/AKT 通路增强 SCAP 的成骨/成牙本质分化能力。
