Objective. Ascorbic acid (AA) and controlled inflammatory stimuli are postulated to possess the ability to independently exert positive effects on a variety of proliferative, pluripotency, and differentiation attributes of gingival mesenchymal stem/progenitor cells (G-MSCs). The current study’s objective was to explore and compare for the first time the impact of the major inflammatory cytokines (IL-1β/TNF-α/IFN-γ), AA, or their combination on multipotency/pluripotency, proliferative, and differentiation characteristics of G-MSCs. Design. Human G-MSCs () were isolated and cultured in basic medium (control group), in basic medium with major inflammatory cytokines; 1 ng/ml IL-1β, 10 ng/ml TNF-α, and 100 ng/ml IFN-γ (inflammatory group), in basic medium with 250 μmol/l AA (AA group) and in inflammatory medium supplemented by AA (inflammatory/AA group). All media were renewed three times per week. In stimulated G-MSCs intracellular β-catenin at 1 hour, pluripotency gene expression at 1, 3, and 5 days, as well as colony-forming units (CFUs) ability and cellular proliferation over 14 days were examined. Following a five-days stimulation in the designated groups, multilineage differentiation was assessed via qualitative and quantitative histochemistry as well as mRNA expression. Results. β-Catenin significantly decreased intracellularly in all experimental groups (, Friedman). AA group exhibited significantly higher cellular counts on days 3, 6, 7, and 13 () and the highest CFUs at 14 days [median-CFUs (Q25/Q75); 40 (15/50), ]. Significantly higher Nanog expression was noted in AA group [median gene-copies/PGK1 (Q25/Q75); 0.0006 (0.0002/0.0007), , Wilcoxon-signed-rank]. Significant multilineage differentiation abilities, especially into osteogenic and chondrogenic directions, were further evident in the AA group. Conclusions. AA stimulation enhances G-MSCs’ stemness, proliferation, and differentiation properties, effects which are associated with a Wnt/β-catenin signaling pathway activation. Apart from initially boosting cellular metabolism as well as Sox2 and Oct4A pluripotency marker expression, inflammation appeared to attenuate these AA-induced positive effects. Current results reveal that for AA to exert its beneficial effects on G-MSCs’ cellular attributes, it requires to act in an inflammation-free microenvironment.

中文翻译：

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抗坏血酸、炎症细胞因子 (IL-1β/TNF-α/IFN-γ) 或其组合对牙龈间充质干/祖细胞干性、增殖和分化的影响。

客观。据推测，抗坏血酸 (AA) 和受控炎症刺激能够独立地对牙龈间充质干细胞/祖细胞 (G-MSCs) 的多种增殖、多能性和分化特性产生积极影响。本研究的目的是首次探索和比较主要炎性细胞因子（IL-1 β /TNF - α /IFN- γ）、AA 或其组合对多能性/多能性、增殖性和分化特征的影响G-MSCs。设计。人类 G-MSCs ()在基础培养基（对照组）中分离培养，在含有主要炎性细胞因子的基础培养基中培养；1 ng/ml IL-1 β、10 ng/ml TNF -α和 100 ng/ml IFN- γ（炎症组），在含有 250 μmol/l AA 的基础培养基 （ AA 组）和补充有AA（炎症/AA组）。所有媒体每周更新 3 次。在受刺激的 G-MSCs 细胞内β检查了 1 小时的 -catenin、1、3 和 5 天的多能性基因表达，以及 14 天内的集落形成单位 (CFU) 能力和细胞增殖。在指定组进行为期五天的刺激后，通过定性和定量组织化学以及 mRNA 表达评估多向分化。结果。在所有实验组中， β-连环蛋白在细胞内显着减少（，弗里德曼）。AA 组在第 3、6、7 和 13 天表现出显着更高的细胞计数（)和 14 天时的最高 CFU [中位 CFU (Q25/Q75)；40 (15/50),]。在 AA 组中注意到显着更高的 Nanog 表达[中位基因拷贝数/PGK1 (Q25/Q75)；0.0006 (0.0002/0.0007),, Wilcoxon 符号秩]。在 AA 组中，显着的多向分化能力，特别是成骨和软骨方向的分化能力更加明显。结论。AA 刺激增强了 G-MSCs 的干性、增殖和分化特性，这些特性与 Wnt/ β-连环蛋白信号通路激活有关。除了最初促进细胞代谢以及 Sox2 和 Oct4A 多能性标志物表达外，炎症似乎减弱了这些 AA 诱导的积极作用。目前的结果表明，AA 要对 G-MSCs 的细胞属性发挥有益作用，需要在无炎症的微环境中发挥作用。