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1. Actomyosin, vimentin and LINC complex pull on osteosarcoma nuclei to deform on micropillar topography

Author

Jean-Louis Milan, Florent Badique, Sebastian Anders, Jean-Noël Freund, Jürgen Rühe, Laurent Pieuchot, Nayana Tusamda Wakhloo, Patricia M. Davidson, Maxime Vassaux, Isabelle Brigaud, Melanie Eichhorn, Tatiana Petithory, Karine Anselme, Institut de Science des Matériaux de Mulhouse (IS2M), Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Department of Microsystems Engineering [Freiburg] (IMTEK), University of Freiburg [Freiburg], Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences du Mouvement Etienne Jules Marey (ISM), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Interface de Recherche Fondamentale et Appliquée en Cancérologie (IRFAC - Inserm U1113), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Paul Strauss : Centre Régional de Lutte contre le Cancer (CRLCC)-Fédération de Médecine Translationelle de Strasbourg (FMTS), Laboratoire Physico-Chimie Curie [Institut Curie] (PCC), Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
LINC complex, Cell, Biophysics, Bone Neoplasms, Bioengineering, Vimentin, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], 02 engineering and technology, Deformation (meteorology), Nucleus, LINC, Biomaterials, 03 medical and health sciences, In vivo, Organelle, medicine, Humans, Cytoskeleton, Migration, 030304 developmental biology, Cell Nucleus, Osteosarcoma, 0303 health sciences, biology, Chemistry, Actomyosin, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, Micropillars, medicine.anatomical_structure, Mechanics of Materials, Ceramics and Composites, biology.protein, 0210 nano-technology, Lamin
Abstract

Cell deformation occurs in many critical biological processes, including cell extravasation during immune response and cancer metastasis. These cells deform the nucleus, its largest and stiffest organelle, while passing through narrow constrictions in vivo and the underlying mechanisms still remain elusive. It is unclear which biochemical actors are responsible and whether the nucleus is pushed or pulled (or both) during deformation. Herein we use an easily-tunable poly-L-lactic acid micropillar topography, mimicking in vivo constrictions to determine the mechanisms responsible for nucleus deformation. Using biochemical tools, we determine that actomyosin contractility, vimentin and nucleo-cytoskeletal connections play essential roles in nuclear deformation, but not A-type lamins. We chemically tune the adhesiveness of the micropillars to show that pulling forces are predominantly responsible for the deformation of the nucleus. We confirm these results using an in silico cell model and propose a comprehensive mechanism for cellular and nuclear deformation during confinement. These results indicate that microstructured biomaterials are extremely versatile tools to understand how forces are exerted in biological systems and can be useful to dissect and mimic complex in vivo behaviour.

Published
2019

2. Tuning InAs Nanowire Density for HEK293 Cell Viability, Adhesion, and Morphology: Perspectives for Nanowire-Based Biosensors

Author

Xiaomei Li, Florent Badique, Karine Anselme, Tor Kristian Andersen, Lei Guo, Nina Buch-Månson, Morten Hannibal Madsen, Jesper Nygård, Sara Bonde, Trine Berthing, and Karen L. Martinez

Subjects
Materials science, Cell Survival, Nanowires, Surface Properties, Nanowire, Nanotechnology, Biosensing Techniques, Adhesion, Cell morphology, Indium, Arsenicals, chemistry.chemical_compound, HEK293 Cells, chemistry, Cell Adhesion, Humans, General Materials Science, Nanorod, Nanotopography, Indium arsenide, Cell adhesion, Nanopillar
Abstract

Arrays of nanowires (NWs) are currently being established as vehicles for molecule delivery and electrical- and fluorescence-based platforms in the development of biosensors. It is conceivable that NW-based biosensors can be optimized through increased understanding of how the nanotopography influences the interfaced biological material. Using state-of-the-art homogenous NW arrays allow for a systematic investigation of how the broad range of NW densities used by the community influences cells. Here it is demonstrated that indium arsenide NW arrays provide a cell-promoting surface, which affects both cell division and focal adhesion up-regulation. Furthermore, a systematic variation in NW spacing affects both the detailed cell morphology and adhesion properties, where the latter can be predicted based on changes in free-energy states using the proposed theoretical model. As the NW density influences cellular parameters, such as cell size and adhesion tightness, it will be important to take NW density into consideration in the continued development of NW-based platforms for cellular applications, such as molecule delivery and electrical measurements.

Published
2013

3. Biomimetic Cryptic Site Surfaces for Reversible Chemo- and Cyto-Mechanoresponsive Substrates

Author

Marie-France Vallat, Benoît Frisch, Florent Badique, Ahmad Fahs, Grégory Francius, Jalal Bacharouche, Maria Vittoria Spanedda, Joseph Hemmerlé, Pierre Schaaf, Jean-Pierre Malval, Karine Anselme, Alexandre Geissler, Vincent Roucoules, Institut de Science des Matériaux de Mulhouse (IS2M), Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie Physique et Microbiologie pour l'Environnement (LCPME), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Biomatériaux et Bioingénierie (BB), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Streptavidin, Materials science, Surface Properties, General Physics and Astronomy, 02 engineering and technology, Cyto-responsive surface, 010402 general chemistry, 01 natural sciences, Mechanotransduction, Cellular, Responsive PEG brush, chemistry.chemical_compound, Biotin, Biomimetic Materials, Mechanochemistry, Elastic Modulus, PEG ratio, Materials Testing, [CHIM]Chemical Sciences, General Materials Science, Mechanoresponsive surface, Mechanotransduction, Ligand, General Engineering, Proteins, Membranes, Artificial, Adhesion, 021001 nanoscience & nanotechnology, 0104 chemical sciences, chemistry, Biochemistry, Biophysics, Stress, Mechanical, 0210 nano-technology, Cryptic site surface, Ethylene glycol, Protein Binding
Abstract

International audience; Chemo-mechanotransduction, the way by which mechanical forces are transformed into chemical signals, plays a fundamental role in many biological processes. The first step of mechanotransduction often relies on exposure, under stretching, of cryptic sites buried in adhesion proteins. Likewise, here we report the first example of synthetic surfaces allowing for specific and fully reversible adhesion of proteins or cells promoted by mechanical action. Silicone sheets are first plasma treated and then functionalized by grafting sequentially under stretching poly(ethylene glycol) (PEG) chains and biotin or arginine-glycine-aspartic acid (RGD) peptides. At unstretched position, these ligands are not accessible for their receptors. Under a mechanical deformation, the surface becomes specifically interactive to streptavidin, biotin antibodies, or adherent for cells, the interactions both for proteins and cells being fully reversible by stretching/unstretching, revealing a reversible exposure process of the ligands. By varying the degree of stretching, the amount of interacting proteins can be varied continuously.

Published
2013

4. Directing nuclear deformation on micropillared surfaces by substrate geometry and cytoskeleton organization

Author

Mathieu Veuillet, Florent Badique, Patricia M. Davidson, Clemens M. Franz, Günter Reiter, Jean-Noël Freund, Dimitar R. Stamov, Karine Anselme, Laboratoire de Bioimagerie et Pathologies (LBP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de Science des Matériaux de Mulhouse (IS2M), Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
Materials science, Cancer cells, Cytoskeleton organization, Polymers, Surface Properties, Polyesters, Biophysics, Bioengineering, Geometry, [SDV.CAN]Life Sciences [q-bio]/Cancer, 02 engineering and technology, Deformation (meteorology), Microscopy, Atomic Force, Biomaterials, 03 medical and health sciences, Cell deformation, Cell Line, Tumor, medicine, Humans, Dimethylpolysiloxanes, Lactic Acid, Cytoskeleton, AFM (atomic force microscopy), Actin, 030304 developmental biology, 0303 health sciences, Force spectroscopy, technology, industry, and agriculture, 021001 nanoscience & nanotechnology, Actin cytoskeleton, Cell biology, Confocal microscopy, Cell nucleus, medicine.anatomical_structure, Mechanics of Materials, Ceramics and Composites, 0210 nano-technology, Nucleus
Abstract

We have recently demonstrated strong nuclear deformation of SaOs-2 osteosarcoma cells on poly-L-lactic acid (PLLA) micropillar substrates. In the present study, we first demonstrated that chemical and mechanical properties of the micropillar substrates have no dominant effect on deformation. However, SaOs-2 nucleus deformation could be strongly modulated by varying the pillar size and spacing, highlighting the importance of geometric constraints for shaping the nucleus. Furthermore, comparing the capacity for nuclear deformation in three different osteosarcoma cell lines (SaOs-2, MG-63 and OHS-4) revealed strong cell-type specific differences. Surprisingly, the highly-deformable SaOs-2 cell line displayed the highest cell stiffness as assessed by AFM-based colloidal force spectroscopy and featured a more prominent array of actin fibres above the nucleus, suggesting a link between actin-mediated cell stiffness and cell nucleus deformation. In contrast, in MG-63 and OHS-4 cells dense microtubule and vimentin networks seem to facilitate some nuclear deformation even in the absence of a prominent actin cytoskeleton. Together these results suggest that an interaction of all three cytoskeletal elements is needed for efficient nuclear deformation. In conclusion, the dominant parameters influencing nuclear deformation on micropillar substrates are not their material properties but the substrate geometry together with cell phenotype and cytoskeleton organization.

Published
2013

5. Nuclear plasticity of cancerous cells on micropillared surfaces: interest for understanding mechanotransduction of metastatic cells

Author

Nayana Tusamda Wakhlo, Florent Badique, Anders, S., Mélanie Eichhorn, Stamov, Dimitar R., Patricia Davidson, Isabelle Brigaud, Laurent Pieuchot, Freund, J. N., Franz, C. M., Jürgen Rühe, Karine Anselme, Institut de Science des Matériaux de Mulhouse (IS2M), Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and univOAK, Archive ouverte

Subjects
[CHIM.MATE] Chemical Sciences/Material chemistry, [CHIM.MATE]Chemical Sciences/Material chemistry
Abstract

Intervention aussi présentée par Karine Anselme à la Journée MEV de l’Institut Jean Lamour le 13 septembre 2016 Affiliations : a. Institut de Sciences des Matériaux de Mulhouse, Université de Strasbourg, Université de Haute-Alsace, CNRS UMR7361, 68057 Mulhouse, France, b. Department of Microsystems Engineering (IMTEK), Albert-Ludwigs-Universität Freiburg, 79110 Freiburg, Germany, c. DFG-Center for Functional Nanostructures, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany, d. Laboratoire Physico-Chimie Curie, Institut Curie, CNRS UMR168, 75248 Paris, France, e. INSERM UMR-S1113, 3 Avenue Molière, 67200 Strasbourg, France; Several years ago, we have shown that strong deformations of cancerous cell bodies can be obtained when osteosarcoma cells are cultured on micropillared substrates and that these alterations also affect the nucleus shape. These strong deformations of cells and of their organelles which don't affect viability and proliferation draw an analogy with the metastatic process. That is why the understanding of cells mechanics and of the related inner forces in this model is essential. Here, we analyse the nuclear deformation on micropillared surfaces of cancer cells. As the deformability of cells is strongly connected with their cytoskeleton dynamics, we combine live imaging and AFM elasticity measurements with treatment with inhibitors of cytoskeleton and LINC complex molecules to decipher the involvement of each cytoskeleton component in the nucleus deformation. In parallel the kinetics of deformation of cancer cells on micropillars is analysed in relation with the chromatin condensation. Our findings confirm that actomyosin microfilaments associated with intermediate filaments and LINC molecules play a major role in this deformation. Microtubules are not always involved depending of the cell type. We also demonstrate that pulling down forces imparted by the cytoskeleton are necessary to promote deformation by changing the chemistry on top or in between interpillar space of pillars by spin coating cell adhesive (PnBA) and repellent polymers (PDMAA).

6. Nucleo-mechanosensing crosstalk between 3-D pillar topography and cyto-nucleoskeleton

Author

Nayana Tusamda Wakhlo, Florent Badique, Anders, S., Mélanie Eichhorn, Isabelle Brigaud, Laurent Pieuchot, Jürgen Ruhe, Karine Anselme, Institut de Science des Matériaux de Mulhouse (IS2M), Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and univOAK, Archive ouverte

Subjects
[CHIM.MATE] Chemical Sciences/Material chemistry, [CHIM.MATE]Chemical Sciences/Material chemistry
Abstract

Nucleo-mechanosensing crosstalk between 3-D pillar topography and cyto-nucleoskeleton

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