Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro
<i>Background and Objectives</i>: Impaired wound healing represents an unsolved medical issue with a high impact on patients’ quality of life and global health care. Even though hypoxia is a significant limiting factor for wound healing, it reveals stimulating effects in gene and protein...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | Medicina |
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| Online Access: | https://www.mdpi.com/1648-9144/59/4/706 |
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| author | Benedikt Fuchs Alexandra Birt Nicholas Moellhoff Constanze Kuhlmann Riccardo E. Giunta Paul Severin Wiggenhauser |
| author_facet | Benedikt Fuchs Alexandra Birt Nicholas Moellhoff Constanze Kuhlmann Riccardo E. Giunta Paul Severin Wiggenhauser |
| author_sort | Benedikt Fuchs |
| collection | DOAJ |
| description | <i>Background and Objectives</i>: Impaired wound healing represents an unsolved medical issue with a high impact on patients’ quality of life and global health care. Even though hypoxia is a significant limiting factor for wound healing, it reveals stimulating effects in gene and protein expression at cellular levels. In particular, hypoxically treated human adipose tissue-derived stem cells (ASCs) have previously been used to stimulate tissue regeneration. Therefore, we hypothesized that they could promote lymphangiogenesis or angiogenesis. <i>Materials and Methods:</i> Dermal regeneration matrices were seeded with human umbilical vein endothelial cells (HUVECs) or human dermal lymphatic endothelial cells (LECs) that were merged with ASCs. Cultures were maintained for 24 h and 7 days under normoxic or hypoxic conditions. Finally, gene and protein expression were measured regarding subtypes of VEGF, corresponding receptors, and intracellular signaling pathways, especially hypoxia-inducible factor-mediated pathways using multiplex-RT-qPCR and ELISA assays. <i>Results:</i> All cell types reacted to hypoxia with an alteration of gene expression. In particular, vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor B (VEGFB), vascular endothelial growth factor C (VEGFC), vascular endothelial growth factor receptor 1 (VEGFR1/FLT1), vascular endothelial growth factor receptor 2 (VEGFR2/KDR), vascular endothelial growth factor receptor 3 (VEGFR3/FLT4), and prospero homeobox 1 (PROX1) were overexpressed significantly depending on upregulation of hypoxia-inducible factor 1 alpha (HIF-1a). Moreover, co-cultures with ASCs showed a more intense change in gene and protein expression profiles and gained enhanced angiogenic and lymphangiogenic potential. In particular, long-term hypoxia led to continuous stimulation of HUVECs by ASCs. <i>Conclusions:</i> Our findings demonstrated the benefit of hypoxic conditioned ASCs in dermal regeneration concerning angiogenesis and lymphangiogenesis. Even a short hypoxic treatment of 24 h led to the stimulation of LECs and HUVECs in an ASC-co-culture. Long-term hypoxia showed a continuous influence on gene expressions. Therefore, this work emphasizes the supporting effects of hypoxia-conditioned-ASC-loaded collagen scaffolds on wound healing in dermal regeneration. |
| first_indexed | 2024-03-11T04:46:23Z |
| format | Article |
| id | doaj.art-95fb5ba79480489eb18ba8328ae1f585 |
| institution | Directory Open Access Journal |
| issn | 1010-660X 1648-9144 |
| language | English |
| last_indexed | 2024-03-11T04:46:23Z |
| publishDate | 2023-04-01 |
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| spelling | doaj.art-95fb5ba79480489eb18ba8328ae1f5852023-11-17T20:20:49ZengMDPI AGMedicina1010-660X1648-91442023-04-0159470610.3390/medicina59040706Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In VitroBenedikt Fuchs0Alexandra Birt1Nicholas Moellhoff2Constanze Kuhlmann3Riccardo E. Giunta4Paul Severin Wiggenhauser5Division of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, GermanyDivision of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, GermanyDivision of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, GermanyDivision of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, GermanyDivision of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, GermanyDivision of Hand, Plastic and Aesthetic Surgery, University Hospital Ludwig-Maximilians-Universität, Ziemssenstraße 5, 80336 Munich, Germany<i>Background and Objectives</i>: Impaired wound healing represents an unsolved medical issue with a high impact on patients’ quality of life and global health care. Even though hypoxia is a significant limiting factor for wound healing, it reveals stimulating effects in gene and protein expression at cellular levels. In particular, hypoxically treated human adipose tissue-derived stem cells (ASCs) have previously been used to stimulate tissue regeneration. Therefore, we hypothesized that they could promote lymphangiogenesis or angiogenesis. <i>Materials and Methods:</i> Dermal regeneration matrices were seeded with human umbilical vein endothelial cells (HUVECs) or human dermal lymphatic endothelial cells (LECs) that were merged with ASCs. Cultures were maintained for 24 h and 7 days under normoxic or hypoxic conditions. Finally, gene and protein expression were measured regarding subtypes of VEGF, corresponding receptors, and intracellular signaling pathways, especially hypoxia-inducible factor-mediated pathways using multiplex-RT-qPCR and ELISA assays. <i>Results:</i> All cell types reacted to hypoxia with an alteration of gene expression. In particular, vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor B (VEGFB), vascular endothelial growth factor C (VEGFC), vascular endothelial growth factor receptor 1 (VEGFR1/FLT1), vascular endothelial growth factor receptor 2 (VEGFR2/KDR), vascular endothelial growth factor receptor 3 (VEGFR3/FLT4), and prospero homeobox 1 (PROX1) were overexpressed significantly depending on upregulation of hypoxia-inducible factor 1 alpha (HIF-1a). Moreover, co-cultures with ASCs showed a more intense change in gene and protein expression profiles and gained enhanced angiogenic and lymphangiogenic potential. In particular, long-term hypoxia led to continuous stimulation of HUVECs by ASCs. <i>Conclusions:</i> Our findings demonstrated the benefit of hypoxic conditioned ASCs in dermal regeneration concerning angiogenesis and lymphangiogenesis. Even a short hypoxic treatment of 24 h led to the stimulation of LECs and HUVECs in an ASC-co-culture. Long-term hypoxia showed a continuous influence on gene expressions. Therefore, this work emphasizes the supporting effects of hypoxia-conditioned-ASC-loaded collagen scaffolds on wound healing in dermal regeneration.https://www.mdpi.com/1648-9144/59/4/706hypoxiaco-cultureangiogenesislymphangiogenesisprotein expressiongene expression |
| spellingShingle | Benedikt Fuchs Alexandra Birt Nicholas Moellhoff Constanze Kuhlmann Riccardo E. Giunta Paul Severin Wiggenhauser Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro Medicina hypoxia co-culture angiogenesis lymphangiogenesis protein expression gene expression |
| title | Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro |
| title_full | Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro |
| title_fullStr | Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro |
| title_full_unstemmed | Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro |
| title_short | Adipose-Derived Stem Cells Improve Angiogenesis and Lymphangiogenesis in a Hypoxic Dermal Regeneration Model In Vitro |
| title_sort | adipose derived stem cells improve angiogenesis and lymphangiogenesis in a hypoxic dermal regeneration model in vitro |
| topic | hypoxia co-culture angiogenesis lymphangiogenesis protein expression gene expression |
| url | https://www.mdpi.com/1648-9144/59/4/706 |
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