Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels
It has been suggested that mechanical strain may elicit cell differentiation in adult somatic cells through activation of epithelial sodium channels (ENaC). However, such phenomenon has not been previously demonstrated in mesenchymal stromal cells (MSCs). The present study was thus conducted to inve...
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| Format: | Article |
| Language: | English |
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Wiley
2020-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2020/5385960 |
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| author | Hui Yin Nam Malliga Raman Murali Raja Elina Ahmad Belinda Pingguan-Murphy Hanumantha Rao Balaji Raghavendran Tunku Kamarul |
| author_facet | Hui Yin Nam Malliga Raman Murali Raja Elina Ahmad Belinda Pingguan-Murphy Hanumantha Rao Balaji Raghavendran Tunku Kamarul |
| author_sort | Hui Yin Nam |
| collection | DOAJ |
| description | It has been suggested that mechanical strain may elicit cell differentiation in adult somatic cells through activation of epithelial sodium channels (ENaC). However, such phenomenon has not been previously demonstrated in mesenchymal stromal cells (MSCs). The present study was thus conducted to investigate the role of ENaC in human bone marrow-derived MSCs (hMSCs) tenogenic differentiation during uniaxial tensile loading. Passaged-2 hMSCs were seeded onto silicone chambers coated with collagen I and subjected to stretching at 1 Hz frequency and 8% strain for 6, 24, 48, and 72 hours. Analyses at these time points included cell morphology and alignment observation, immunocytochemistry and immunofluorescence staining (collagen I, collagen III, fibronectin, and N-cadherin), and gene expression (ENaC subunits, and tenogenic markers). Unstrained cells at similar time points served as the control group. To demonstrate the involvement of ENaC in the differentiation process, an ENaC blocker (benzamil) was used and the results were compared to the noninhibited hMSCs. ENaC subunits’ (α, β, γ, and δ) expression was observed in hMSCs, although only α subunit was significantly increased during stretching. An increase in tenogenic genes’ (collagen1, collagen3, decorin, tenascin-c, scleraxis, and tenomodulin) and proteins’ (collagen I, collagen III, fibronectin, and N-cadherin) expression suggests that hMSCs underwent tenogenic differentiation when subjected to uniaxial loading. Inhibition of ENaC function resulted in decreased expression of these markers, thereby suggesting that ENaC plays a vital role in tenogenic differentiation of hMSCs during mechanical loading. |
| format | Article |
| id | doaj-art-615859169ee341d9a10e74dac2194642 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-615859169ee341d9a10e74dac21946422025-02-03T06:04:36ZengWileyStem Cells International1687-966X1687-96782020-01-01202010.1155/2020/53859605385960Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium ChannelsHui Yin Nam0Malliga Raman Murali1Raja Elina Ahmad2Belinda Pingguan-Murphy3Hanumantha Rao Balaji Raghavendran4Tunku Kamarul5Tissue Engineering Group, Department of Orthopaedic Surgery (NOCERAL), Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaTissue Engineering Group, Department of Orthopaedic Surgery (NOCERAL), Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaDepartment of Physiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaDepartment of Biomedical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, MalaysiaTissue Engineering Group, Department of Orthopaedic Surgery (NOCERAL), Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaTissue Engineering Group, Department of Orthopaedic Surgery (NOCERAL), Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaIt has been suggested that mechanical strain may elicit cell differentiation in adult somatic cells through activation of epithelial sodium channels (ENaC). However, such phenomenon has not been previously demonstrated in mesenchymal stromal cells (MSCs). The present study was thus conducted to investigate the role of ENaC in human bone marrow-derived MSCs (hMSCs) tenogenic differentiation during uniaxial tensile loading. Passaged-2 hMSCs were seeded onto silicone chambers coated with collagen I and subjected to stretching at 1 Hz frequency and 8% strain for 6, 24, 48, and 72 hours. Analyses at these time points included cell morphology and alignment observation, immunocytochemistry and immunofluorescence staining (collagen I, collagen III, fibronectin, and N-cadherin), and gene expression (ENaC subunits, and tenogenic markers). Unstrained cells at similar time points served as the control group. To demonstrate the involvement of ENaC in the differentiation process, an ENaC blocker (benzamil) was used and the results were compared to the noninhibited hMSCs. ENaC subunits’ (α, β, γ, and δ) expression was observed in hMSCs, although only α subunit was significantly increased during stretching. An increase in tenogenic genes’ (collagen1, collagen3, decorin, tenascin-c, scleraxis, and tenomodulin) and proteins’ (collagen I, collagen III, fibronectin, and N-cadherin) expression suggests that hMSCs underwent tenogenic differentiation when subjected to uniaxial loading. Inhibition of ENaC function resulted in decreased expression of these markers, thereby suggesting that ENaC plays a vital role in tenogenic differentiation of hMSCs during mechanical loading.http://dx.doi.org/10.1155/2020/5385960 |
| spellingShingle | Hui Yin Nam Malliga Raman Murali Raja Elina Ahmad Belinda Pingguan-Murphy Hanumantha Rao Balaji Raghavendran Tunku Kamarul Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels Stem Cells International |
| title | Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels |
| title_full | Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels |
| title_fullStr | Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels |
| title_full_unstemmed | Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels |
| title_short | Mechanical Strain-Mediated Tenogenic Differentiation of Mesenchymal Stromal Cells Is Regulated through Epithelial Sodium Channels |
| title_sort | mechanical strain mediated tenogenic differentiation of mesenchymal stromal cells is regulated through epithelial sodium channels |
| url | http://dx.doi.org/10.1155/2020/5385960 |
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