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Image-based Characterization of 3D Collagen Networks and the Effect of Embedded Cells

Published online by Cambridge University Press:  18 June 2019

Vanesa Olivares ,
Mar Cóndor ,
Cristina Del Amo ,
Jesús Asín ,
Carlos Borau  and
José Manuel García-Aznar Open the ORCID record for José Manuel García-Aznar [Opens in a new window]
Vanesa Olivares
Affiliation:
Multiscale in Mechanical and Biological Engineering (Department of Mechanical Engineering), University of Zaragoza, Zaragoza, Spain Aragon Institute of Engineering Research, University of Zaragoza, Zaragoza, Spain
Mar Cóndor
Affiliation:
Multiscale in Mechanical and Biological Engineering (Department of Mechanical Engineering), University of Zaragoza, Zaragoza, Spain Aragon Institute of Engineering Research, University of Zaragoza, Zaragoza, Spain Biomechanics Section, Department of Mechanical Engineering, KU Leuven, Belgium
Cristina Del Amo
Affiliation:
Multiscale in Mechanical and Biological Engineering (Department of Mechanical Engineering), University of Zaragoza, Zaragoza, Spain Aragon Institute of Engineering Research, University of Zaragoza, Zaragoza, Spain
Jesús Asín
Affiliation:
Department of Statistical Methods, University of Zaragoza, Zaragoza, Spain
Carlos Borau
Affiliation:
Multiscale in Mechanical and Biological Engineering (Department of Mechanical Engineering), University of Zaragoza, Zaragoza, Spain Aragon Institute of Engineering Research, University of Zaragoza, Zaragoza, Spain University Center for Defense, Zaragoza, Spain
José Manuel García-Aznar*
Affiliation:
Multiscale in Mechanical and Biological Engineering (Department of Mechanical Engineering), University of Zaragoza, Zaragoza, Spain Aragon Institute of Engineering Research, University of Zaragoza, Zaragoza, Spain
*
*Author for correspondence: José Manuel García-Aznar, E-mail: [email protected]
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Abstract

Collagen microstructure is closely related to the mechanical properties of tissues and affects cell migration through the extracellular matrix. To study these structures, three-dimensional (3D) in vitro collagen-based gels are often used, attempting to mimic the natural environment of cells. Some key parameters of the microstructure of these gels are fiber orientation, fiber length, or pore size, which define the mechanical properties of the network and therefore condition cell behavior. In the present study, an automated tool to reconstruct 3D collagen networks is used to extract the aforementioned parameters of gels of different collagen concentration and determine how their microstructure is affected by the presence of cells. Two different experiments are presented to test the functionality of the method: first, collagen gels are embedded within a microfluidic device and collagen fibers are imaged by using confocal fluorescence microscopy; second, collagen gels are directly polymerized in a cell culture dish and collagen fibers are imaged by confocal reflection microscopy. Finally, we investigate and compare the collagen microstructure far from and in the vicinities of MDA-MB 23 cells, finding that cell activity during migration was able to strongly modify the orientation of the collagen fibers and the porosity-related values.

Keywords

collagen gelconfocal microscopyextracellular matrixmicrostructurescaffold morphology3D reconstruction
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 25 , Issue 4 , August 2019 , pp. 971 - 981
Copyright
Copyright © Microscopy Society of America 2019 

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