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The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy

The connection between cytoskeleton alterations and diseases is well known and has stimulated research on cell mechanics, aiming to develop reliable biomarkers. In this study, we present results on rheological, adhesion, and morphological properties of primary rat cardiac fibroblasts, the cytoskelet...

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Autores principales: Braidotti, Nicoletta, do R. B. F. Lima, Maria Augusta, Zanetti, Michele, Rubert, Alessandro, Ciubotaru, Catalin, Lazzarino, Marco, Sbaizero, Orfeo, Cojoc, Dan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9029771/
https://www.ncbi.nlm.nih.gov/pubmed/35456926
http://dx.doi.org/10.3390/ijms23084108
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author Braidotti, Nicoletta
do R. B. F. Lima, Maria Augusta
Zanetti, Michele
Rubert, Alessandro
Ciubotaru, Catalin
Lazzarino, Marco
Sbaizero, Orfeo
Cojoc, Dan
author_facet Braidotti, Nicoletta
do R. B. F. Lima, Maria Augusta
Zanetti, Michele
Rubert, Alessandro
Ciubotaru, Catalin
Lazzarino, Marco
Sbaizero, Orfeo
Cojoc, Dan
author_sort Braidotti, Nicoletta
collection PubMed
description The connection between cytoskeleton alterations and diseases is well known and has stimulated research on cell mechanics, aiming to develop reliable biomarkers. In this study, we present results on rheological, adhesion, and morphological properties of primary rat cardiac fibroblasts, the cytoskeleton of which was altered by treatment with cytochalasin D (Cyt-D) and nocodazole (Noc), respectively. We used two complementary techniques: quartz crystal microbalance (QCM) and digital holographic microscopy (DHM). Qualitative data on cell viscoelasticity and adhesion changes at the cell–substrate near-interface layer were obtained with QCM, while DHM allowed the measurement of morphological changes due to the cytoskeletal alterations. A rapid effect of Cyt-D was observed, leading to a reduction in cell viscosity, loss of adhesion, and cell rounding, often followed by detachment from the surface. Noc treatment, instead, induced slower but continuous variations in the rheological behavior for four hours of treatment. The higher vibrational energy dissipation reflected the cell’s ability to maintain a stable attachment to the substrate, while a cytoskeletal rearrangement occurs. In fact, along with the complete disaggregation of microtubules at prolonged drug exposure, a compensatory effect of actin polymerization emerged, with increased stress fiber formation.
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spelling pubmed-90297712022-04-23 The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy Braidotti, Nicoletta do R. B. F. Lima, Maria Augusta Zanetti, Michele Rubert, Alessandro Ciubotaru, Catalin Lazzarino, Marco Sbaizero, Orfeo Cojoc, Dan Int J Mol Sci Article The connection between cytoskeleton alterations and diseases is well known and has stimulated research on cell mechanics, aiming to develop reliable biomarkers. In this study, we present results on rheological, adhesion, and morphological properties of primary rat cardiac fibroblasts, the cytoskeleton of which was altered by treatment with cytochalasin D (Cyt-D) and nocodazole (Noc), respectively. We used two complementary techniques: quartz crystal microbalance (QCM) and digital holographic microscopy (DHM). Qualitative data on cell viscoelasticity and adhesion changes at the cell–substrate near-interface layer were obtained with QCM, while DHM allowed the measurement of morphological changes due to the cytoskeletal alterations. A rapid effect of Cyt-D was observed, leading to a reduction in cell viscosity, loss of adhesion, and cell rounding, often followed by detachment from the surface. Noc treatment, instead, induced slower but continuous variations in the rheological behavior for four hours of treatment. The higher vibrational energy dissipation reflected the cell’s ability to maintain a stable attachment to the substrate, while a cytoskeletal rearrangement occurs. In fact, along with the complete disaggregation of microtubules at prolonged drug exposure, a compensatory effect of actin polymerization emerged, with increased stress fiber formation. MDPI 2022-04-07 /pmc/articles/PMC9029771/ /pubmed/35456926 http://dx.doi.org/10.3390/ijms23084108 Text en © 2022 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
Braidotti, Nicoletta
do R. B. F. Lima, Maria Augusta
Zanetti, Michele
Rubert, Alessandro
Ciubotaru, Catalin
Lazzarino, Marco
Sbaizero, Orfeo
Cojoc, Dan
The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title_full The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title_fullStr The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title_full_unstemmed The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title_short The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy
title_sort role of cytoskeleton revealed by quartz crystal microbalance and digital holographic microscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9029771/
https://www.ncbi.nlm.nih.gov/pubmed/35456926
http://dx.doi.org/10.3390/ijms23084108
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