Cargando…
Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells
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 we...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699152/ https://www.ncbi.nlm.nih.gov/pubmed/34943818 http://dx.doi.org/10.3390/cells10123310 |
_version_ | 1784620448396345344 |
---|---|
author | 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. |
author_facet | 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. |
author_sort | Fawzy El-Sayed, Karim M. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8699152 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-86991522021-12-24 Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells 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. Cells Article 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. MDPI 2021-11-25 /pmc/articles/PMC8699152/ /pubmed/34943818 http://dx.doi.org/10.3390/cells10123310 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article 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. Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title | Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title_full | Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title_fullStr | Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title_full_unstemmed | Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title_short | Ascorbic Acid/Retinol and/or Inflammatory Stimuli’s Effect on Proliferation/Differentiation Properties and Transcriptomics of Gingival Stem/Progenitor Cells |
title_sort | ascorbic acid/retinol and/or inflammatory stimuli’s effect on proliferation/differentiation properties and transcriptomics of gingival stem/progenitor cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8699152/ https://www.ncbi.nlm.nih.gov/pubmed/34943818 http://dx.doi.org/10.3390/cells10123310 |
work_keys_str_mv | AT fawzyelsayedkarimm ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT bittneramira ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT schlichtkristina ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT mekhemarmohamed ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT enthammerkim ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT hoppnermarc ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT essounimartha ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT schulzjuliane ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT laudesmatthias ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT graetzchristian ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT dorferchristofe ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells AT schultedominikm ascorbicacidretinolandorinflammatorystimuliseffectonproliferationdifferentiationpropertiesandtranscriptomicsofgingivalstemprogenitorcells |