Ascorbic Acid/Retinol and/or Inflammatory Stimuli's Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells

• Published in: Cells (Basel, Switzerland) Switzerland), 2021-11, Vol.10 (12), p.3310
• Main Authors: Fawzy El-Sayed, Karim M, Bittner, Amira, Schlicht, Kristina, Mekhemar, Mohamed, Enthammer, Kim, Höppner, Marc, Es-Souni, Martha, Schulz, Juliane, Laudes, Matthias, Graetz, Christian, Dörfer, Christof E, Schulte, Dominik M
• Format: Article
• Language: English
• Subjects: Adolescent; Antigen processing; Ascorbic acid; Ascorbic Acid - pharmacology; beta Catenin - metabolism; Biomarkers - metabolism; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cell Lineage - drug effects; Cell Lineage - genetics; Cell proliferation; Cell Proliferation - drug effects; Cell Proliferation - genetics; Colony-Forming Units Assay; EGR-1 protein; Experiments; Female; Gene Expression Profiling; Gene Expression Regulation - drug effects; Genes; gingiva; Gingiva - pathology; Humans; IL-1β; Inflammation; Inflammation - genetics; Inflammation - pathology; Intracellular; Intracellular levels; Major histocompatibility complex; Male; Mesenchymal Stem Cells - drug effects; Mesenchymal Stem Cells - metabolism; Mesenchymal Stem Cells - pathology; Mesenchyme; Next-generation sequencing; Period 1 protein; Progenitor cells; retinol; RNA, Messenger - genetics; RNA, Messenger - metabolism; Software; stem cell; Stem cells; Tissue Donors; Transcriptome - drug effects; Transcriptome - genetics; Transcriptomics; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Vitamin A; Vitamin A - pharmacology; Young Adult; β-Catenin; γ-Interferon
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells10123310

The present study explored the effects of ascorbic-acid (AA)/retinol and timed inflammation on the stemness, the regenerative potential, and the transcriptomics profile of gingival mesenchymal stem/progenitor cells' (G-MSCs). STRO-1 (mesenchymal stem cell marker) immuno-magnetically sorted G-MSCs were cultured in basic medium (control group), in basic medium with IL-1β (1 ng/mL), TNF-α (10 ng/mL) and IFN-γ (100 ng/mL, inflammatory-medium), in basic medium with AA (250 µmol/L) and retinol (20 µmol/L) (AA/retinol group) or in inflammatory medium with AA/retinol (inflammatory/AA/retinol group; = 5/group). The intracellular levels of phosphorylated and total β-Catenin at 1 h, the expression of stemness genes over 7 days, the number of colony-forming units (CFUs) as well as the cellular proliferation aptitude over 14 days, and the G-MSCs' multilineage differentiation potential were assessed. Next-generation sequencing was undertaken to elaborate on up-/downregulated genes and altered intracellular pathways. G-MSCs demonstrated all mesenchymal stem/progenitor cells characteristics. Controlled inflammation with AA/retinol significantly elevated ( < 0.05). The AA/retinol-mediated reduction in intracellular phosphorylated β-Catenin was restored through the effect of controlled inflammation ( < 0.05). Cellular proliferation was highest in the AA/retinol group ( < 0.05). AA/retinol counteracted the inflammation-mediated reduction in G-MSCs' clonogenic ability and CFUs. Amplified chondrogenic differentiation was observed in the inflammatory/AA/retinol group. At 1 and 3 days, the differentially expressed genes were associated with development, proliferation, and migration ( , , , , , , ), survival ( , , ), differentiation and mineral absorption ( , , , , , ), inflammation and MHC-II antigen processing ( , , ) and intracellular pathway activation ( , ). Less as well as more genes were activated the longer the G-MSCs remained in the inflammatory medium or AA/retinol, respectively. Combined, current results point at possibly interesting interactions between controlled inflammation or AA/retinol affecting stemness, proliferation, and differentiation attributes of G-MSCs.