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            <funder>This work was supported by ANR-23-CE19-0025 (FibersForCells) and by ANR-JCJC InfEx (ANR-20-CE15-0023). This work received support from the grant Q-life ANR-17-CONV-0005 and from the grants ANR-11-LABX-0038, ANR-10-IDEX-0001-02 [LabEx Cell(n)Scale] and ANR-10-IDEX-0001-02 PSL. This work was supported by a doctoral grant from the program “Interfaces pour le Vivant,” Sorbonne Université (recipient P.U.), a doctoral grant for PhD 4th year from la Ligue contre le Cancer (recipient P.U.), a doctoral grant from the Oversea Study Program of Guangzhou Elite Project (recipient X.J.), a doctoral grant from Université Paris Sciences et Lettres (PSL), Thèses binômées (recipients I.C. and E.N.). This work has received the support of “Institut Pierre-Gilles de Gennes” laboratoire d’excellence, “Investissements d’avenir” (program ANR-10-IDEX-0001-02, ANR-10-EQPX-34 and ANR-10-LABX-31). This work was supported by the Centre National de la Recherche Scientifique (CNRS), Sorbonne Université, PSL University and Institut Curie. This work has received support under the program France 2030 launched by the French Government.</funder>
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                <title level="j">Proceedings of the National Academy of Sciences of the United States of America</title>
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                <term xml:lang="en">3D traction forces</term>
                <term xml:lang="en">cell contractility</term>
                <term xml:lang="en">Atomic Force Microscopy</term>
                <term xml:lang="en">fibers</term>
                <term xml:lang="en">Two-Photon Polymerization</term>
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              <p>The forces exerted by cells upon the fibers of the extracellular matrix play a decisive role in cell motility in physiopathology. How the local physical properties of the matrix (density, stiffness, orientation) affect cellular forces remains, however, poorly understood. Existing approaches to measure cell three-dimensional (3D) traction forces within fibrous substrates lack control over the local properties and rely on continuum approaches, not suited for measuring forces at the scale of individual fibers. Herein, an approach is proposed to fabricate multilayer arrays of suspended deformable fibers spanning a wide range of fine-tunable geometrical and mechanical properties using two-photon polymerization. Atomic Force Microscopy is used to thoroughly investigate the properties of individual fibers, including Young’s modulus and stiffness. This approach is combined with a reference-free method for measuring traction forces in 3D, which relies on automated segmentation of the fibers coupled with finite element modeling. The force measurement pipeline is applied to study forces exerted by endothelial cells, fibroblasts, or macrophages, and reveals how these forces are influenced by fiber density and stiffness. Additionally, coupling to fast volumetric imaging with lattice light-sheet microscopy enables the measurement of the low-intensity and short-lived tractions exerted by amoeboid cells, such as dendritic cells. Our technology will be instrumental for monitoring and studying cell behavior at the single-fiber level at extracellular matrix density interfaces, which play a crucial role in both physiological and pathological contexts, such as tumor boundaries.</p>
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