Research Article Ascorbic Acid, Inflammatory Cytokines (IL-1β/TNF-α/IFN-γ), or Their Combination’s Effect on Stemness, Proliferation, and Differentiation of Gingival Mesenchymal Stem/Progenitor Cells Karim M. Fawzy El-Sayed , 1,2,3 Nhung Nguyen, 2 and Christof E. Dörfer 2 1 Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo, Egypt 2 Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian Albrechts University, Kiel, Germany 3 Oral Medicine and Periodontology Department, Faculty of Dentistry, Cairo University, Egypt Correspondence should be addressed to Karim M. Fawzy El-Sayed; karim.fawzy@gmail.com Received 1 May 2020; Revised 16 July 2020; Accepted 30 July 2020; Published 17 August 2020 Academic Editor: De-Meng Chen Copyright © 2020 Karim M. Fawzy El-Sayed et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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 (n =5) 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 (p =0:002, Friedman). AA group exhibited significantly higher cellular counts on days 3, 6, 7, and 13 (p <0:05) and the highest CFUs at 14 days [median-CFUs (Q25/Q75); 40 (15/50), p =0:043]. Significantly higher Nanog expression was noted in AA group [median gene-copies/PGK1 (Q25/Q75); 0.0006 (0.0002/0.0007), p <0:01, 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. 1. Introduction Periodontitis is an inflammatory disorder of tooth-investing and tooth-supporting tissues, branded by a gradual damage of alveolar bone, periodontal attachment, and eventually gin- giva, associated with bacterial dysbiosis. The commencement of this multifaceted disease process commonly entails challeng- ing of the periodontal immune-inflammatory system through virulent microbial biofilms. Subsequently, an inflammatory reaction is mounted, with the release of inflammatory cyto- kines, most prominently tumor necrosis factor alpha (TNF- α), interleukin (IL) 1 beta (IL-1β), IL-4, IL-6, and interferon gamma (IFN-γ) [1]. Duration and intensity of the resultant host reaction govern the personalized course and outcome of the inflammatory process, as well as affect the outcome of any subsequent periodontal reparative/regenerative approach. Hindawi Stem Cells International Volume 2020, Article ID 8897138, 14 pages https://doi.org/10.1155/2020/8897138