Cargando…
Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope
Mechanical properties of healthy and Dupuytren fibroblasts were investigated by atomic force microscopy (AFM). In addition to standard force curves, rheological properties were assessed using an oscillatory testing methodology, in which the frequency was swept from 1 Hz to 1 kHz, and data were analy...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917339/ https://www.ncbi.nlm.nih.gov/pubmed/36768366 http://dx.doi.org/10.3390/ijms24032043 |
_version_ | 1784886341961515008 |
---|---|
author | Pérez-Domínguez, Sandra López-Alonso, Javier Lafont, Frank Radmacher, Manfred |
author_facet | Pérez-Domínguez, Sandra López-Alonso, Javier Lafont, Frank Radmacher, Manfred |
author_sort | Pérez-Domínguez, Sandra |
collection | PubMed |
description | Mechanical properties of healthy and Dupuytren fibroblasts were investigated by atomic force microscopy (AFM). In addition to standard force curves, rheological properties were assessed using an oscillatory testing methodology, in which the frequency was swept from 1 Hz to 1 kHz, and data were analyzed using the structural damping model. Dupuytren fibroblasts showed larger apparent Young’s modulus values than healthy ones, which is in agreement with previous results. Moreover, cell mechanics were compared before and after ML-7 treatment, which is a myosin light chain kinase inhibitor (MLCK) that reduces myosin activity and hence cell contraction. We employed two different concentrations of ML-7 inhibitor and could observe distinct cell reactions. At 1 µM, healthy and scar fibroblasts did not show measurable changes in stiffness, but Dupuytren fibroblasts displayed a softening and recovery after some time. When increasing ML-7 concentration (3 µM), the majority of cells reacted, Dupuytren fibroblasts were the most susceptible, not being able to recover from the drug and dying. These results suggested that ML-7 is a potent inhibitor for MLCK and that myosin II is essential for cytoskeleton stabilization and cell survival. |
format | Online Article Text |
id | pubmed-9917339 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99173392023-02-11 Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope Pérez-Domínguez, Sandra López-Alonso, Javier Lafont, Frank Radmacher, Manfred Int J Mol Sci Article Mechanical properties of healthy and Dupuytren fibroblasts were investigated by atomic force microscopy (AFM). In addition to standard force curves, rheological properties were assessed using an oscillatory testing methodology, in which the frequency was swept from 1 Hz to 1 kHz, and data were analyzed using the structural damping model. Dupuytren fibroblasts showed larger apparent Young’s modulus values than healthy ones, which is in agreement with previous results. Moreover, cell mechanics were compared before and after ML-7 treatment, which is a myosin light chain kinase inhibitor (MLCK) that reduces myosin activity and hence cell contraction. We employed two different concentrations of ML-7 inhibitor and could observe distinct cell reactions. At 1 µM, healthy and scar fibroblasts did not show measurable changes in stiffness, but Dupuytren fibroblasts displayed a softening and recovery after some time. When increasing ML-7 concentration (3 µM), the majority of cells reacted, Dupuytren fibroblasts were the most susceptible, not being able to recover from the drug and dying. These results suggested that ML-7 is a potent inhibitor for MLCK and that myosin II is essential for cytoskeleton stabilization and cell survival. MDPI 2023-01-20 /pmc/articles/PMC9917339/ /pubmed/36768366 http://dx.doi.org/10.3390/ijms24032043 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Pérez-Domínguez, Sandra López-Alonso, Javier Lafont, Frank Radmacher, Manfred Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title | Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title_full | Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title_fullStr | Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title_full_unstemmed | Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title_short | Comparison of Rheological Properties of Healthy versus Dupuytren Fibroblasts When Treated with a Cell Contraction Inhibitor by Atomic Force Microscope |
title_sort | comparison of rheological properties of healthy versus dupuytren fibroblasts when treated with a cell contraction inhibitor by atomic force microscope |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917339/ https://www.ncbi.nlm.nih.gov/pubmed/36768366 http://dx.doi.org/10.3390/ijms24032043 |
work_keys_str_mv | AT perezdominguezsandra comparisonofrheologicalpropertiesofhealthyversusdupuytrenfibroblastswhentreatedwithacellcontractioninhibitorbyatomicforcemicroscope AT lopezalonsojavier comparisonofrheologicalpropertiesofhealthyversusdupuytrenfibroblastswhentreatedwithacellcontractioninhibitorbyatomicforcemicroscope AT lafontfrank comparisonofrheologicalpropertiesofhealthyversusdupuytrenfibroblastswhentreatedwithacellcontractioninhibitorbyatomicforcemicroscope AT radmachermanfred comparisonofrheologicalpropertiesofhealthyversusdupuytrenfibroblastswhentreatedwithacellcontractioninhibitorbyatomicforcemicroscope |