Hwa Somyeong, Lee Hyun-Jin, Ko Youngkyung, Park Jun-Beom
Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Dental Implantology, Graduate School of Clinical Dental Science, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Medicina (Kaunas). 2025 Jan 4;61(1):76. doi: 10.3390/medicina61010076.
Insulin-like growth factor-1 (IGF-1) plays a vital role in various cellular processes, including those involving stem cells. This study evaluated the effects of IGF-1 on cell survival, osteogenic differentiation, and mRNA expression in gingiva-derived mesenchymal stem cell spheroids. Using concave microwells, spheroids were generated in the presence of IGF-1 at concentrations of 0, 10, and 100 ng/mL. Cellular vitality was qualitatively assessed using microscopy, while a water-soluble tetrazolium salt-based assay kit quantified cellular viability. Osteogenic differentiation was evaluated via alkaline phosphatase activity and an anthraquinone dye test to measure calcium deposition. Additionally, quantitative polymerase chain reaction (qPCR) analysis was performed to determine the expression of RUNX2 and COL1A1. By day 1, the stem cell spheroids had successfully formed, and their morphology remained stable over the following 7 days. The IGF-1 concentrations tested showed no significant differences in cell viability. Similarly, alkaline phosphatase activity on day 7 revealed no observable changes. However, on day 7, the incorporation of IGF-1 led to an increase in Alizarin Red staining, indicative of enhanced calcium deposition. Notably, an IGF-1 concentration of 100 ng/mL significantly upregulated the expression of COL1A1. These findings suggest that IGF-1 supports the maintenance of cell viability and promotes the expression of COL1A1 in gingiva-derived mesenchymal stem cell spheroids, highlighting its potential role in enhancing osteogenic differentiation. Future research should include long-term studies to evaluate the sustainability of IGF-1-induced effects on stem cell spheroids.
胰岛素样生长因子-1(IGF-1)在包括涉及干细胞的各种细胞过程中发挥着至关重要的作用。本研究评估了IGF-1对牙龈来源的间充质干细胞球体的细胞存活、成骨分化和mRNA表达的影响。使用凹形微孔板,在浓度为0、10和100 ng/mL的IGF-1存在下生成球体。使用显微镜对细胞活力进行定性评估,同时使用基于水溶性四氮唑盐的检测试剂盒对细胞活力进行定量。通过碱性磷酸酶活性和蒽醌染料试验评估成骨分化,以测量钙沉积。此外,进行定量聚合酶链反应(qPCR)分析以确定RUNX2和COL1A1的表达。到第1天,干细胞球体已成功形成,并且在接下来的7天内其形态保持稳定。所测试的IGF-1浓度在细胞活力方面没有显示出显著差异。同样,第7天的碱性磷酸酶活性没有观察到变化。然而,在第7天,添加IGF-1导致茜素红染色增加,表明钙沉积增强。值得注意的是,100 ng/mL的IGF-1浓度显著上调了COL1A1的表达。这些发现表明,IGF-1支持牙龈来源的间充质干细胞球体的细胞活力维持,并促进COL1A1的表达,突出了其在增强成骨分化中的潜在作用。未来的研究应包括长期研究,以评估IGF-1对干细胞球体诱导作用的可持续性。