Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells
Skeletal muscle-derived stem cells (MDSCs) are the key modulators of muscle regeneration. An inappropriate cellular microenvironment can reduce the regenerative capacity of MDSCs. This study evaluates the effect of microenvironmental alterations on the cell differentiation capacity using either mech...
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MDPI AG
2024-04-01
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| author | Paula Hawlitschek Michele C. Klymiuk Asmaa Eldaey Sabine Wenisch Stefan Arnhold Mohamed I. Elashry |
| author_facet | Paula Hawlitschek Michele C. Klymiuk Asmaa Eldaey Sabine Wenisch Stefan Arnhold Mohamed I. Elashry |
| author_sort | Paula Hawlitschek |
| collection | DOAJ |
| description | Skeletal muscle-derived stem cells (MDSCs) are the key modulators of muscle regeneration. An inappropriate cellular microenvironment can reduce the regenerative capacity of MDSCs. This study evaluates the effect of microenvironmental alterations on the cell differentiation capacity using either mechanical fluid shear stress (FSS) or hypoxic conditions. C2C12 mouse myoblasts were differentiated under cyclic FSS (CFSS), periodic FSS (PFSS) for one hour, and hypoxia (3% O<sub>2</sub>) for up to seven days. Cell proliferation and myogenic differentiation capacities were evaluated using cell viability assays, immunohistochemical staining, and morphometric analysis. The expression of MyoD, myogenin, myosin heavy chain, nitric oxide, hypoxia-inducible factor 1 alpha (HIF1α), vascular endothelial growth factor (VEGF) and mammalian target of rapamycin (mTOR) was quantified by means of RT-qPCR. The data showed that FSS conditions altered cell morphology and increased cell viability and cell distribution compared to static conditions. MyoD and myogenin expression was upregulated under both FSS conditions. CFSS induction improved myogenic differentiation parameters including myotube number, size and fusion capacity. Although hypoxia enhanced cell viability compared to normoxia, it reduced differentiation capacity, as indicated by the downregulation of myogenin and mTOR expression, as well as reducing myotube formation. Under hypoxic conditions, increased nitric oxide production and upregulation of VEGF expression were detected for up to 72 h. The data suggest an improved myogenic differentiation capacity under mechanical FSS; in contrast, the cell differentiation capacity was impaired under hypoxic conditions. The data point out that optimizing the biomechanical and oxidative stressors in the cellular microenvironment could improve stem cell transplantation and enhance their regenerative potential in the context of cell-based therapies. |
| first_indexed | 2024-04-24T10:48:28Z |
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| last_indexed | 2024-04-24T10:48:28Z |
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| spelling | doaj.art-10d99e23763b43ed975b81d181f31f652024-04-12T13:15:32ZengMDPI AGApplied Sciences2076-34172024-04-01147304710.3390/app14073047Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem CellsPaula Hawlitschek0Michele C. Klymiuk1Asmaa Eldaey2Sabine Wenisch3Stefan Arnhold4Mohamed I. Elashry5Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanyInstitute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanyClinic of Small Animals, c/o Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanyClinic of Small Animals, c/o Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanyInstitute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanyInstitute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, GermanySkeletal muscle-derived stem cells (MDSCs) are the key modulators of muscle regeneration. An inappropriate cellular microenvironment can reduce the regenerative capacity of MDSCs. This study evaluates the effect of microenvironmental alterations on the cell differentiation capacity using either mechanical fluid shear stress (FSS) or hypoxic conditions. C2C12 mouse myoblasts were differentiated under cyclic FSS (CFSS), periodic FSS (PFSS) for one hour, and hypoxia (3% O<sub>2</sub>) for up to seven days. Cell proliferation and myogenic differentiation capacities were evaluated using cell viability assays, immunohistochemical staining, and morphometric analysis. The expression of MyoD, myogenin, myosin heavy chain, nitric oxide, hypoxia-inducible factor 1 alpha (HIF1α), vascular endothelial growth factor (VEGF) and mammalian target of rapamycin (mTOR) was quantified by means of RT-qPCR. The data showed that FSS conditions altered cell morphology and increased cell viability and cell distribution compared to static conditions. MyoD and myogenin expression was upregulated under both FSS conditions. CFSS induction improved myogenic differentiation parameters including myotube number, size and fusion capacity. Although hypoxia enhanced cell viability compared to normoxia, it reduced differentiation capacity, as indicated by the downregulation of myogenin and mTOR expression, as well as reducing myotube formation. Under hypoxic conditions, increased nitric oxide production and upregulation of VEGF expression were detected for up to 72 h. The data suggest an improved myogenic differentiation capacity under mechanical FSS; in contrast, the cell differentiation capacity was impaired under hypoxic conditions. The data point out that optimizing the biomechanical and oxidative stressors in the cellular microenvironment could improve stem cell transplantation and enhance their regenerative potential in the context of cell-based therapies.https://www.mdpi.com/2076-3417/14/7/3047skeletal musclestem cellsfluid shear stresshypoxiamyogenic differentiation |
| spellingShingle | Paula Hawlitschek Michele C. Klymiuk Asmaa Eldaey Sabine Wenisch Stefan Arnhold Mohamed I. Elashry Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells Applied Sciences skeletal muscle stem cells fluid shear stress hypoxia myogenic differentiation |
| title | Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells |
| title_full | Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells |
| title_fullStr | Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells |
| title_full_unstemmed | Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells |
| title_short | Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells |
| title_sort | modulation of the cellular microenvironment by mechanical fluid shear stress and hypoxia alters the differentiation capacity of skeletal muscle derived stem cells |
| topic | skeletal muscle stem cells fluid shear stress hypoxia myogenic differentiation |
| url | https://www.mdpi.com/2076-3417/14/7/3047 |
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