| Autor: |
Fawzy El-Sayed KM; Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany.; Oral Medicine and Periodontology Department, Faculty of Dentistry, Cairo University, Cairo 11553, Egypt.; Stem cells and Tissue Engineering Unit, Faculty of Dentistry, Cairo University, Cairo 11553, Egypt., Bittner A; Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Schlicht K; Institute of Diabetes and Clinical Metabolic Research, School of Medicine, Christian-Albrechts-University of Kiel, 24104 Kiel, Germany.; Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Medicine I, School of Medicine, University Hospital of Schleswig-Holstein, 24105 Kiel, Germany., Mekhemar M; Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Enthammer K; Institute of Diabetes and Clinical Metabolic Research, School of Medicine, Christian-Albrechts-University of Kiel, 24104 Kiel, Germany.; Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Medicine I, School of Medicine, University Hospital of Schleswig-Holstein, 24105 Kiel, Germany., Höppner M; Institute of Clinical Molecular Biology, School of Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Es-Souni M; Department of Orthodontics, School of Dental Medicine, University Clinic Schleswig-Holstein (UKSH), Christian-Albrechts University of Kiel, 24105 Kiel, Germany., Schulz J; Institute of Diabetes and Clinical Metabolic Research, School of Medicine, Christian-Albrechts-University of Kiel, 24104 Kiel, Germany.; Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Medicine I, School of Medicine, University Hospital of Schleswig-Holstein, 24105 Kiel, Germany.; Cluster of Excellence, Precision Medicine in Chronic Inflammation, School of Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Laudes M; Institute of Diabetes and Clinical Metabolic Research, School of Medicine, Christian-Albrechts-University of Kiel, 24104 Kiel, Germany.; Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Medicine I, School of Medicine, University Hospital of Schleswig-Holstein, 24105 Kiel, Germany.; Cluster of Excellence, Precision Medicine in Chronic Inflammation, School of Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Graetz C; Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Dörfer CE; Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany., Schulte DM; Institute of Diabetes and Clinical Metabolic Research, School of Medicine, Christian-Albrechts-University of Kiel, 24104 Kiel, Germany.; Division of Endocrinology, Diabetes and Clinical Nutrition, Department of Medicine I, School of Medicine, University Hospital of Schleswig-Holstein, 24105 Kiel, Germany.; Cluster of Excellence, Precision Medicine in Chronic Inflammation, School of Medicine, Christian-Albrechts-University of Kiel, 24105 Kiel, Germany. |
| Abstrakt: |
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; n = 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 NANOG ( p < 0.05). The AA/retinol-mediated reduction in intracellular phosphorylated β-Catenin was restored through the effect of controlled inflammation ( p < 0.05). Cellular proliferation was highest in the AA/retinol group ( p < 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 ( FOS , EGR1 , SGK1 , CXCL5 , SIPA1L2 , TFPI2 , KRATP1-5 ), survival ( EGR1 , SGK1 , TMEM132A ), differentiation and mineral absorption ( FOS , EGR1 , MT1E , KRTAP1-5 , ASNS , PSAT1 ), inflammation and MHC-II antigen processing ( PER1 , CTSS , CD74 ) and intracellular pathway activation ( FKBP5 , ZNF404 ). 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. |
