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

Abstract

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.

Retinol-Loaded Poly(vinyl alcohol)-Based Hydrogels as Suitable Biomaterials with Antimicrobial Properties for the Proliferation of Mesenchymal Stem Cells

“…It was also stated that retinol affects the differentiation potential of gingival progenitor stem cells through activating Wnt/β-catenin signals, and the differentiation of MSCs into neural cells could be achieved by retinol treatment through upregulating the expression of vimentin, Stra13, and RARα/β/γ and downregulating Brachyury expression [91]. To support the present finding, a recent study by Fawzy et al reported the regenerative potential effect of ascorbic acid retinol using gingival mesenchymal stem/progenitor cells, and their study concluded that the proliferative and chondrogenic differentiation of MSCs by ascorbic acid and retinol was triggered by activating phosphorylated β-Catenin, FOS, EGR1, SGK1, CXCL5, SIPA1L2, TFPI2, KRATP1-5, MT1E, ASNS, and PSAT1 gene expression in gingival mesenchymal stem cells [88]. Comparatively, the cell-loading ability of PVA hydrogel was similar to retinol-loaded PVA hydrogel, which ultimately proved the efficiency of the PVA retinol hydrogel system in the stem cell culture.…”

“…The BMMS cells’ proliferative effect of PVA hydrogel was significantly improved by the incorporation of retinol from 0.1 to 0.5 wt.%. Several studies already claimed the beneficial effect of retinol in stem cell proliferation, metabolism, and differentiation [ 86 , 87 , 88 , 89 , 90 , 91 ]. The differentiation potential of mesenchymal stem cells originated from mouse ciliary epithelium into retinal neurons by retinol treatment and could be achieved through elevating the expression of the rhodopsin protein and the Nestin, RPE65, and Rhodopsin genes [ 90 ].…”

Can vitamins improve periodontal wound healing/regeneration?

“…136 Furthermore, AA boosts the telomerase activity and pluripotency of stem/progenitor cells, osteoblasts and fibroblasts. [137][138][139][140][141][142] AA stimulates fibroblast migration and keratinocyte proliferation, with the potential for improving the epithelial phenotype. 143 It functions as an enzymatic cofactor for a large family of metalloenzymes involved in the synthesis and stabilization of collagen of the periodontal ligament, gingiva, cement and alveolar bone, 144,145 in the production of peptide hormones and neurotransmitters, and in the regulation of transcription factors, such as hypoxia inducible factor-1.…”

“…AA is believed to support wound healing through pleiotropic mechanisms, including the promotion of neovascularization, VEGF expression, extracellular matrix deposition, cellular renewal, and differentiation, while inhibiting cellular apoptosis 136 . Furthermore, AA boosts the telomerase activity and pluripotency of stem/progenitor cells, osteoblasts and fibroblasts 137–142 . AA stimulates fibroblast migration and keratinocyte proliferation, with the potential for improving the epithelial phenotype 143 .…”

“…In a further in vitro investigation, G‐MSCs were combinedly stimulated by AA and retinol under an uninflamed as well as an experimental inflammatory setup. Results revealed that AA/retinol reversed the inflammation‐mediated decrease in G‐MSCs' clonogenic ability and CFUs, amplified chondrogenic differentiation potential, with a higher activation of genes 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 ) 142 . Again, in this study a conjoint application of micronutrient appeared to be beneficial (Table 23).…”