Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model
Unsuccessful wound closure in chronic wounds can be linked to altered keratinocyte activation and their inability to re-epithelize. Suggested mechanisms driving this impairment involve unbalanced cytokine signaling. However, the molecular events leading to these aberrant responses are poorly underst...
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MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/7/6210 |
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| author | Sergio Liarte Ángel Bernabé-García Mónica Rodríguez-Valiente José M. Moraleda Gregorio Castellanos Francisco J. Nicolás |
| author_facet | Sergio Liarte Ángel Bernabé-García Mónica Rodríguez-Valiente José M. Moraleda Gregorio Castellanos Francisco J. Nicolás |
| author_sort | Sergio Liarte |
| collection | DOAJ |
| description | Unsuccessful wound closure in chronic wounds can be linked to altered keratinocyte activation and their inability to re-epithelize. Suggested mechanisms driving this impairment involve unbalanced cytokine signaling. However, the molecular events leading to these aberrant responses are poorly understood. Among cytokines affecting keratinocyte responses, Transforming Growth Factor-β (TFG-β) is thought to have a great impact. In this study, we have used a previously characterized skin epidermal in vitro model, HaCaT cells continuously exposed to TGF-β1, to study the wound recovery capabilities of chronified/senescent keratinocytes. In this setting, chronified keratinocytes show decreased migration and reduced activation in response to injury. Amniotic membrane (AM) has been used successfully to manage unresponsive complicated wounds. In our in vitro setting, AM treatment of chronified keratinocytes re-enabled migration in the early stages of wound healing, also promoting proliferation at later stages. Interestingly, when checking the gene expression of markers known to be altered in TGF-β chronified cells and involved in cell cycle regulation, early migratory responses, senescence, and chronic inflammation, we discovered that AM treatment seemed to reset back to keratinocyte status. The analysis of the evolution of both the levels of keratinocyte activation marker cytokeratin 17 and the spatial-temporal expression pattern of the proliferation marker Ki-67 in human in vivo biopsy samples suggests that responses to AM recorded in TGF-β chronified HaCaT cells would be homologous to those of resident keratinocytes in chronic wounds. All these results provide further evidence that sustained TGF-β might play a key role in wound chronification and postulate the validity of our TGF-β chronified HaCaT in vitro model for the study of chronic wound physiology. |
| first_indexed | 2024-03-11T05:36:11Z |
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| id | doaj.art-c6908bcf6c034dd980ed1a945ff3640b |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T05:36:11Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c6908bcf6c034dd980ed1a945ff3640b2023-11-17T16:47:55ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-03-01247621010.3390/ijms24076210Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell ModelSergio Liarte0Ángel Bernabé-García1Mónica Rodríguez-Valiente2José M. Moraleda3Gregorio Castellanos4Francisco J. Nicolás5Laboratorio de Regeneración, Oncología Molecular y TGF-β, IMIB-Arrixaca, El Palmar, 30120 Murcia, SpainLaboratorio de Regeneración, Oncología Molecular y TGF-β, IMIB-Arrixaca, El Palmar, 30120 Murcia, SpainLaboratorio de Regeneración, Oncología Molecular y TGF-β, IMIB-Arrixaca, El Palmar, 30120 Murcia, SpainUnidad de Terapia Celular, Hospital Clínico Universitario Virgen de la Arrixaca, El Palmar, 30120 Murcia, SpainUnidad de Heridas Crónicas y Úlcera de Pie Diabético, Hospital Clínico Universitario Virgen de la Arrixaca, El Palmar, 30120 Murcia, SpainLaboratorio de Regeneración, Oncología Molecular y TGF-β, IMIB-Arrixaca, El Palmar, 30120 Murcia, SpainUnsuccessful wound closure in chronic wounds can be linked to altered keratinocyte activation and their inability to re-epithelize. Suggested mechanisms driving this impairment involve unbalanced cytokine signaling. However, the molecular events leading to these aberrant responses are poorly understood. Among cytokines affecting keratinocyte responses, Transforming Growth Factor-β (TFG-β) is thought to have a great impact. In this study, we have used a previously characterized skin epidermal in vitro model, HaCaT cells continuously exposed to TGF-β1, to study the wound recovery capabilities of chronified/senescent keratinocytes. In this setting, chronified keratinocytes show decreased migration and reduced activation in response to injury. Amniotic membrane (AM) has been used successfully to manage unresponsive complicated wounds. In our in vitro setting, AM treatment of chronified keratinocytes re-enabled migration in the early stages of wound healing, also promoting proliferation at later stages. Interestingly, when checking the gene expression of markers known to be altered in TGF-β chronified cells and involved in cell cycle regulation, early migratory responses, senescence, and chronic inflammation, we discovered that AM treatment seemed to reset back to keratinocyte status. The analysis of the evolution of both the levels of keratinocyte activation marker cytokeratin 17 and the spatial-temporal expression pattern of the proliferation marker Ki-67 in human in vivo biopsy samples suggests that responses to AM recorded in TGF-β chronified HaCaT cells would be homologous to those of resident keratinocytes in chronic wounds. All these results provide further evidence that sustained TGF-β might play a key role in wound chronification and postulate the validity of our TGF-β chronified HaCaT in vitro model for the study of chronic wound physiology.https://www.mdpi.com/1422-0067/24/7/6210chronic woundscell modelsamniotic membranewound healingkeratinocytesTGF-β |
| spellingShingle | Sergio Liarte Ángel Bernabé-García Mónica Rodríguez-Valiente José M. Moraleda Gregorio Castellanos Francisco J. Nicolás Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model International Journal of Molecular Sciences chronic wounds cell models amniotic membrane wound healing keratinocytes TGF-β |
| title | Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model |
| title_full | Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model |
| title_fullStr | Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model |
| title_full_unstemmed | Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model |
| title_short | Amniotic Membrane Restores Chronic Wound Features to Normal in a Keratinocyte TGF-β-Chronified Cell Model |
| title_sort | amniotic membrane restores chronic wound features to normal in a keratinocyte tgf β chronified cell model |
| topic | chronic wounds cell models amniotic membrane wound healing keratinocytes TGF-β |
| url | https://www.mdpi.com/1422-0067/24/7/6210 |
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