Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study
Human mesenchymal stem cells (MSC) are multipotent stem cells, which are isolated from various sources. Currently, there is a worldwide interest for dental MSC to be used against neurodegenerative diseases, since they derive from the neural crest and express embryonic stem cell markers. This fact pr...
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MDPI AG
2022-01-01
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| author | Anastasios E. Koutsoumparis Anastasia Patsiarika Anastasia Tsingotjidou Ioannis Pappas Asterios S. Tsiftsoglou |
| author_facet | Anastasios E. Koutsoumparis Anastasia Patsiarika Anastasia Tsingotjidou Ioannis Pappas Asterios S. Tsiftsoglou |
| author_sort | Anastasios E. Koutsoumparis |
| collection | DOAJ |
| description | Human mesenchymal stem cells (MSC) are multipotent stem cells, which are isolated from various sources. Currently, there is a worldwide interest for dental MSC to be used against neurodegenerative diseases, since they derive from the neural crest and express embryonic stem cell markers. This fact prompted us to explore their potential for neural trans-differentiation in culture. We employed all-trans-retinoic acid (ATRA) and 2-(3-ethylureido)-6-methylpyridine (UDP-4) to induce neural differentiation of human MSC from the dental apical papilla (SCAP). The SCAP were exposed to either agent separately and assessed for proliferation, viability, morphology, and gene expression of the following neural-specific markers: neuron-specific enolase (<i>ENO2</i>), neurofibromin 1 (<i>NF1</i>), choline acetyltransferase (<i>CHAT</i>), tyrosine hydroxylase (<i>TH</i>), and the vesicular GABA transporter (<i>SLC32A1</i>). They were also assessed for the expression of glial fibrillary acidic protein (GFAP) and neuronal nuclear antigen (NeuN) by immunofluorescence. ATRA or UDP-4 treatment inhibited the cell growth and promoted limited cell death, but to a different extent. The addition of the neuroprotective agent recombinant human erythropoietin-alpha (rhEPO-α) enhanced the UDP-4-inducing capacity for more than three weeks. ATRA or UDP-4 treatment significantly upregulated <i>ENO2</i> and <i>NF1</i> expression, indicating neuronal differentiation. Moreover, the ATRA treatment significantly induced the upregulation of the GABAergic-specific <i>SLC32A1</i>, while the UDP-4 treatment led to the significant upregulation of the adrenergic-specific <i>TH</i>. The UDP-4 treatment induced the expression of NeuN and GFAP after four and three weeks, respectively, while the ATRA-treatment did not. Our findings indicate that SCAP can be differentiated into neural-like cells after treatment with ATRA or UDP-4 by exhibiting a disparate pattern of differentiation. Therefore, UDP-4 is suggested here as a new potent neural-differentiation-inducing compound, which, when combined with rhEPO-α, could lay the foundation for robust stem-cell-based therapies of neurodegeneration. |
| first_indexed | 2024-03-09T22:29:54Z |
| format | Article |
| id | doaj.art-7281ac65c91149ada0b6e747ec298f1f |
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| language | English |
| last_indexed | 2024-03-09T22:29:54Z |
| publishDate | 2022-01-01 |
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| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-7281ac65c91149ada0b6e747ec298f1f2023-11-23T18:58:23ZengMDPI AGBiomolecules2218-273X2022-01-0112221810.3390/biom12020218Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative StudyAnastasios E. Koutsoumparis0Anastasia Patsiarika1Anastasia Tsingotjidou2Ioannis Pappas3Asterios S. Tsiftsoglou4Laboratory of Pharmacology, Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 54124 Thessaloniki, GreeceLaboratory of Pharmacology, Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 54124 Thessaloniki, GreeceLaboratory of Anatomy, Histology and Embryology, Faculty of Health Sciences, School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, GreeceLaboratory of Pharmacology and Toxicology, Faculty of Veterinary Science, University of Thessaly, 43100 Karditsa, GreeceLaboratory of Pharmacology, Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 54124 Thessaloniki, GreeceHuman mesenchymal stem cells (MSC) are multipotent stem cells, which are isolated from various sources. Currently, there is a worldwide interest for dental MSC to be used against neurodegenerative diseases, since they derive from the neural crest and express embryonic stem cell markers. This fact prompted us to explore their potential for neural trans-differentiation in culture. We employed all-trans-retinoic acid (ATRA) and 2-(3-ethylureido)-6-methylpyridine (UDP-4) to induce neural differentiation of human MSC from the dental apical papilla (SCAP). The SCAP were exposed to either agent separately and assessed for proliferation, viability, morphology, and gene expression of the following neural-specific markers: neuron-specific enolase (<i>ENO2</i>), neurofibromin 1 (<i>NF1</i>), choline acetyltransferase (<i>CHAT</i>), tyrosine hydroxylase (<i>TH</i>), and the vesicular GABA transporter (<i>SLC32A1</i>). They were also assessed for the expression of glial fibrillary acidic protein (GFAP) and neuronal nuclear antigen (NeuN) by immunofluorescence. ATRA or UDP-4 treatment inhibited the cell growth and promoted limited cell death, but to a different extent. The addition of the neuroprotective agent recombinant human erythropoietin-alpha (rhEPO-α) enhanced the UDP-4-inducing capacity for more than three weeks. ATRA or UDP-4 treatment significantly upregulated <i>ENO2</i> and <i>NF1</i> expression, indicating neuronal differentiation. Moreover, the ATRA treatment significantly induced the upregulation of the GABAergic-specific <i>SLC32A1</i>, while the UDP-4 treatment led to the significant upregulation of the adrenergic-specific <i>TH</i>. The UDP-4 treatment induced the expression of NeuN and GFAP after four and three weeks, respectively, while the ATRA-treatment did not. Our findings indicate that SCAP can be differentiated into neural-like cells after treatment with ATRA or UDP-4 by exhibiting a disparate pattern of differentiation. Therefore, UDP-4 is suggested here as a new potent neural-differentiation-inducing compound, which, when combined with rhEPO-α, could lay the foundation for robust stem-cell-based therapies of neurodegeneration.https://www.mdpi.com/2218-273X/12/2/218mesenchymal stem cells (MSC)stem cells from the apical papilla (SCAP)all-trans-retinoic acid (ATRA)2-(3-ethylureido)-6-methylpyridine (UDP-4)neural trans-differentiationimmunofluorescence |
| spellingShingle | Anastasios E. Koutsoumparis Anastasia Patsiarika Anastasia Tsingotjidou Ioannis Pappas Asterios S. Tsiftsoglou Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study Biomolecules mesenchymal stem cells (MSC) stem cells from the apical papilla (SCAP) all-trans-retinoic acid (ATRA) 2-(3-ethylureido)-6-methylpyridine (UDP-4) neural trans-differentiation immunofluorescence |
| title | Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study |
| title_full | Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study |
| title_fullStr | Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study |
| title_full_unstemmed | Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study |
| title_short | Neural Differentiation of Human Dental Mesenchymal Stem Cells Induced by ATRA and UDP-4: A Comparative Study |
| title_sort | neural differentiation of human dental mesenchymal stem cells induced by atra and udp 4 a comparative study |
| topic | mesenchymal stem cells (MSC) stem cells from the apical papilla (SCAP) all-trans-retinoic acid (ATRA) 2-(3-ethylureido)-6-methylpyridine (UDP-4) neural trans-differentiation immunofluorescence |
| url | https://www.mdpi.com/2218-273X/12/2/218 |
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