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Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells

In the course of the development of the nervous system, neuronal cells extend (grow) axons, which navigate over distances of the order of many cell diameters to reach target dendrites from other neurons and establish neuronal circuits. Some of the central challenges in biophysics today are to develo...

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Autores principales: Kumarasinghe, Udathari, Fox, Lucian N., Staii, Cristian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9624305/
https://www.ncbi.nlm.nih.gov/pubmed/36278714
http://dx.doi.org/10.3390/biomimetics7040157
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author Kumarasinghe, Udathari
Fox, Lucian N.
Staii, Cristian
author_facet Kumarasinghe, Udathari
Fox, Lucian N.
Staii, Cristian
author_sort Kumarasinghe, Udathari
collection PubMed
description In the course of the development of the nervous system, neuronal cells extend (grow) axons, which navigate over distances of the order of many cell diameters to reach target dendrites from other neurons and establish neuronal circuits. Some of the central challenges in biophysics today are to develop a quantitative model of axonal growth, which includes the interactions between the neurons and their growth environment, and to describe the complex architecture of neuronal networks in terms of a small number of physical variables. To address these challenges, researchers need new experimental techniques for measuring biomechanical interactions with very high force and spatiotemporal resolutions. Here we report a unique experimental approach that integrates three different high-resolution techniques on the same platform—traction force microscopy (TFM), atomic force microscopy (AFM) and fluorescence microscopy (FM)—to measure biomechanical properties of cortical neurons. To our knowledge, this is the first literature report of combined TFM/AFM/FM measurements performed for any type of cell. Using this combination of powerful experimental techniques, we perform high-resolution measurements of the elastic modulus for cortical neurons and relate these values with traction forces exerted by the cells on the growth substrate (poly acrylamide hydrogels, or PAA, coated with poly D-lysine). We obtain values for the traction stresses exerted by the cortical neurons in the range 30–70 Pa, and traction forces in the range 5–11 nN. Our results demonstrate that neuronal cells stiffen when axons exert forces on the PAA substrate, and that neuronal growth is governed by a contact guidance mechanism, in which axons are guided by external mechanical cues. This work provides new insights for bioengineering novel biomimetic platforms that closely model neuronal growth in vivo, and it has significant impact for creating neuroprosthetic interfaces and devices for neuronal growth and regeneration.
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spelling pubmed-96243052022-11-02 Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells Kumarasinghe, Udathari Fox, Lucian N. Staii, Cristian Biomimetics (Basel) Article In the course of the development of the nervous system, neuronal cells extend (grow) axons, which navigate over distances of the order of many cell diameters to reach target dendrites from other neurons and establish neuronal circuits. Some of the central challenges in biophysics today are to develop a quantitative model of axonal growth, which includes the interactions between the neurons and their growth environment, and to describe the complex architecture of neuronal networks in terms of a small number of physical variables. To address these challenges, researchers need new experimental techniques for measuring biomechanical interactions with very high force and spatiotemporal resolutions. Here we report a unique experimental approach that integrates three different high-resolution techniques on the same platform—traction force microscopy (TFM), atomic force microscopy (AFM) and fluorescence microscopy (FM)—to measure biomechanical properties of cortical neurons. To our knowledge, this is the first literature report of combined TFM/AFM/FM measurements performed for any type of cell. Using this combination of powerful experimental techniques, we perform high-resolution measurements of the elastic modulus for cortical neurons and relate these values with traction forces exerted by the cells on the growth substrate (poly acrylamide hydrogels, or PAA, coated with poly D-lysine). We obtain values for the traction stresses exerted by the cortical neurons in the range 30–70 Pa, and traction forces in the range 5–11 nN. Our results demonstrate that neuronal cells stiffen when axons exert forces on the PAA substrate, and that neuronal growth is governed by a contact guidance mechanism, in which axons are guided by external mechanical cues. This work provides new insights for bioengineering novel biomimetic platforms that closely model neuronal growth in vivo, and it has significant impact for creating neuroprosthetic interfaces and devices for neuronal growth and regeneration. MDPI 2022-10-08 /pmc/articles/PMC9624305/ /pubmed/36278714 http://dx.doi.org/10.3390/biomimetics7040157 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
Kumarasinghe, Udathari
Fox, Lucian N.
Staii, Cristian
Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title_full Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title_fullStr Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title_full_unstemmed Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title_short Combined Traction Force–Atomic Force Microscopy Measurements of Neuronal Cells
title_sort combined traction force–atomic force microscopy measurements of neuronal cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9624305/
https://www.ncbi.nlm.nih.gov/pubmed/36278714
http://dx.doi.org/10.3390/biomimetics7040157
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