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2. Combining fluorescence fluctuations and photobleaching to quantify surface density

Author

Sefkow-Werner, Julius, Migliorini, Elisa, Picart, Catherine, Wahyuni, Dwiria, Wang, Irene, and Delon, Antoine

Subjects
Physics - Biological Physics
Abstract

We establish a method, called pbFFS for photobleaching Fluctuation Fluorescence Spectroscopy, which aims at characterizing molecules or particles labelled with a unknown distribution or fluorophores. Using photobleaching as a control parameter, pbFFS provides information on the distribution of labels and a more reliable estimation of the absolute density or concentration of these molecules. We present a complete theoretical derivation of the pbFFS approach and experimentally apply it to measure the surface density of a monolayer of fluorescently tagged streptavidin molecules that can be used as a base platform for biomimetic systems. The surface density measured by pbFFS is consistent with the results of more standard surface techniques, such as ellipsometry. Compared to those techniques, pbFFS has two main advantages: it enables in situ characterization (no dedicated substrates are required) and is applicable to low masses of adsorbed molecules, as demonstrated here by quantifying the density of biotin-Atto molecules that bind to the streptavidin layer. Besides molecules immobilized on surfaces, we also applied pbFFS to molecules diffusing in solution, to confirm the distribution of labels found on surfaces. Hence, pbFFS provides a set of tools to investigate molecules attached to a variable number of fluorophores, with the aim toquantify either the number of molecules or the distribution of fluorophores, the latter case being especially relevant for oligomerization studies., Comment: 31 pages, 7 figures and supporting information (4 pages, including 2 figures)

Published
2021

5. Differential structural remodelling of heparan sulfate by chemokines: the role of chemokine oligomerization

Author

Dyer, Douglas P, Migliorini, Elisa, Salanga, Catherina L, Thakar, Dhruv, Handel, Tracy M, and Richter, Ralf P

Subjects
Biochemistry and Cell Biology, Biological Sciences, Aetiology, 2.1 Biological and endogenous factors, Underpinning research, 1.1 Normal biological development and functioning, Animals, Binding Sites, Cell Adhesion, Chemokines, Heparitin Sulfate, Humans, Models, Molecular, Mutation, Protein Binding, Protein Conformation, Protein Multimerization, Receptors, Chemokine, Signal Transduction, chemokine, glycosaminoglycan, heparan sulfate, extracellular matrix, glycocalyx, quartz crystal microbalance with dissipation monitoring, Microbiology, Immunology, Biological sciences, Biomedical and clinical sciences
Abstract

Chemokines control the migration of cells in normal physiological processes and in the context of disease such as inflammation, autoimmunity and cancer. Two major interactions are involved: (i) binding of chemokines to chemokine receptors, which activates the cellular machinery required for movement; and (ii) binding of chemokines to glycosaminoglycans (GAGs), which facilitates the organization of chemokines into haptotactic gradients that direct cell movement. Chemokines can bind and activate their receptors as monomers; however, the ability to oligomerize is critical for the function of many chemokines in vivo Chemokine oligomerization is thought to enhance their affinity for GAGs, and here we show that it significantly affects the ability of chemokines to accumulate on and be retained by heparan sulfate (HS). We also demonstrate that several chemokines differentially rigidify and cross-link HS, thereby affecting HS rigidity and mobility, and that HS cross-linking is significantly enhanced by chemokine oligomerization. These findings suggest that chemokine-GAG interactions may play more diverse biological roles than the traditional paradigms of physical immobilization and establishment of chemokine gradients; we hypothesize that they may promote receptor-independent events such as physical re-organization of the endothelial glycocalyx and extracellular matrix, as well as signalling through proteoglycans to facilitate leukocyte adhesion and transmigration.

Published
2017

11. Glycosaminoglycans Exhibit Distinct Interactions and Signaling with Bmp2 According to Their Nature and Localization

Author

Le Pennec, Jean, primary, Makshakova, Olga N., additional, Nevola, Paola, additional, Fouladkar, Farah, additional, Gout, Evelyne, additional, Machillot, Paul, additional, Friedel-Arboleas, Mélanie, additional, Picart, Catherine, additional, Perez, Serge, additional, Vortkamp, Andrea, additional, Vivès, Romain R., additional, and Migliorini, Elisa, additional

Published
2024
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12. BMP-Binding Polysulfonate Brushes to Control Growth Factor Presentation and Regulate Matrix Remodelling

Author

Marchena, Metzli Hernandez, Lambert, Elisa, Bogdanović, Bojana, Quadir, Fauzia, Neri-Cruz, Carlos E., Luo, Jiajun, Nadal, Clemence, Migliorini, Elisa, and Gautrot, Julien E.

Abstract

Bone morphogenetic proteins (BMPs) are important targets to incorporate in biomaterial scaffolds to orchestrate tissue repair. Glycosaminoglycans (GAGs) such as heparin allow the capture of BMPs and their retention at the surface of biomaterials at safe concentrations. Although heparin has strong affinities for BMP2 and BMP4, two important types of growth factors regulating bone and tissue repair, it remains difficult to embed stably at the surface of a broad range of biomaterials and degrades rapidly in vitro and in vivo. In this report, biomimetic poly(sulfopropyl methacrylate) (PSPMA) brushes are proposed as sulfated GAG mimetic interfaces for the stable capture of BMPs. The growth of PSPMA brushes via a surface-initiated activator regenerated by electron transfer polymerization is investigated via ellipsometry, prior to characterization of swelling and surface chemistry via X-ray photoelectron spectroscopy and Fourier transform infrared. The capacity of PSPMA brushes to bind BMP2 and BMP4 is then characterized via surface plasmon resonance. BMP2 is found to anchor particularly stably and at high density at the surface of PSPMA brushes, and a strong impact of the brush architecture on binding capacity is observed. These results are further confirmed using a quartz crystal microbalance with dissipation monitoring, providing some insights into the mode of adsorption of BMPs at the surface of PSPMA brushes. Primary adsorption of BMP2, with relatively little infiltration, is observed on thick dense brushes, implying that this growth factor should be accessible for further binding of corresponding cell membrane receptors. Finally, to demonstrate the impact of PSPMA brushes for BMP2 capture, dermal fibroblasts were then cultured at the surface of functionalized PSPMA brushes. The presence of BMP2 and the architecture of the brush are found to have a significant impact on matrix deposition at the corresponding interfaces. Therefore, PSPMA brushes emerge as attractive coatings for scaffold engineering and stable capture of BMP2 for regenerative medicine applications.

Published
2024
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19. Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Author

Sefkow-Werner, Julius, primary, Le Pennec, Jean, additional, Machillot, Paul, additional, Ndayishimiye, Bertin, additional, Castro-Ramirez, Elaine, additional, Lopes, João, additional, Licitra, Christophe, additional, Wang, Irene, additional, Delon, Antoine, additional, Picart, Catherine, additional, and Migliorini, Elisa, additional

Published
2022
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25. High throughput measurements of bone morphogenetic protein (BMP)/BMP receptors interactions using bio-layer interferometry

Author

Khodr, Valia, Machillot, Paul, Migliorini, Elisa, Reiser, Jean-Baptiste, and Picart, Catherine

Subjects
Bone Morphogenetic Protein 7, Bone Morphogenetic Protein 2, Biosensing Techniques, Bone Morphogenetic Protein 4, Bone Morphogenetic Protein Receptors, Surface Plasmon Resonance, Bone Morphogenetic Protein Receptors, Type II, Article, Kinetics, Interferometry, Bone Morphogenetic Proteins, Growth Differentiation Factor 2, Humans, Dimerization, Bone Morphogenetic Protein Receptors, Type I, Protein Binding
Abstract

Bone morphogenetic proteins (BMP) are an important family of growth factors playing a role in a large number of physiological and pathological processes, including bone homeostasis, tissue regeneration and cancers. In vivo, BMPs bind successively to both BMP receptors (BMPR) of type I and type II, and a promiscuity has been reported. In this study, we used bio-layer interferometry to perform parallel real-time biosensing and to deduce the kinetic parameters (k(a), k(d)) and the equilibrium constant (K(D)) for a large range of BMPs/BMPR combinations in similar experimental conditions. We selected four members of the BMP family (BMP-2, 4, 7, 9) known for their physiological relevance and studied their interactions with five type-I BMP receptors (ALK1, 2, 3, 5, 6) and three type-II BMP receptors (BMPR-II, ACTR-IIA, ACTR-IIB). We reveal that BMP-2 and BMP-4 behave differently, especially regarding their kinetic interactions and affinities with the type-II BMPR. We found that BMP-7 has a higher affinity for the type-II BMPR receptor ACTR-IIA and a tenfold lower affinity with the type-I receptors. While BMP-9 has a high and similar affinity for all type-II receptors, it can interact with ALK5 and ALK2, in addition to ALK1. Interestingly, we also found that all BMPs can interact with ALK5. The interaction between BMPs and both type-I and type II receptors in a ternary complex did not reveal further cooperativity. Our work provides a synthetic view of the interactions of these BMPs with their receptors and paves the way for future studies on their cell-type and receptor specific signaling pathways.

Published
2021

30. Citation

Author

Cavalcanti-Adam, Elisabetta Ada, Migliorini, Elisa, Laboratoire des matériaux et du génie physique (LMGP ), and Institut National Polytechnique de Grenoble (INPG)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, [CHIM.POLY]Chemical Sciences/Polymers, [SDV]Life Sciences [q-bio], [SDV.BDD.MOR]Life Sciences [q-bio]/Development Biology/Morphogenesis, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019
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35. Nanostructured substrates to control the Embryonic Stem cells differentiation into neuronal lineage

Author

Migliorini, Elisa, Lazzarino, Marco, Janmey, Paul, and Cecchini, Marco

Subjects
FIS/01 FISICA SPERIMENTALE, mechanotrasduction, Nanopatterned substrates, cell adhesion, SCUOLA DI DOTTORATO DI RICERCA IN NANOTECNOLOGIE, Embyonic stem cells, neuronal differentiation
Abstract

2010/2011 The objective of this project was to develop new nanotechnology-based strategies to increase embryonic stem cells (ESCs) differentiation into neuronal lineage. In particular it was chosen to investigate a nanostructured physical support for in vitro stem cell culture in which both the nanometrical topography and mechanical properties are well controlled and characterized. Nanopatterned substrates were designed to have physical properties as close as possible to the in vivo microenvironment where stem cells normally grow and differentiate based on the assumption that mimicking the natural niche equilibrium is of fundamental importance for stem cell fate. First, an original nanotechnological approach to fabricate the substrates for in vitro neuronal precursors culture was developed. Secondly the substrate geometrical and mechanical parameters were optimized in order to achieve the maximum differentiation yield of ESCs-derived neuronal precursors (NPs). It was reached a neuronal yield of 74±7% at 48 hours after NPs differentiation induction, which represents the highest yield ever published using nanopatterned substrates with controlled and highthroughput reproducible nanometrical features for cell culture. Moreover it was demonstrated that the mechanical properties of the substrate play a major role with respect to other parameters, such as substrate composition and geometry. A time-dependent analysis showed that the first hours after cell seeding are crucial in the determination of the final differentiation yield. A further control of ESCs differentiation by manipulating the substrates physical parameters, required a deep understanding of the cell-substrate interaction, therefore it was studied the behavior of neuronal precursors when placed and grown on different artificial substrates using atomic force microscope, scanning electron microscope, and single cell force spectroscopy measurements. The latter lead to a quantification of the forces that develop between neuronal precursors and substrate and provided a clear relationship between adhesion forces and differentiation. My results suggested the importance of the physical parameter involved in the regulation of the neuronal differentiation and to new guidelines for future applications in regenerative medicine. XXIV Ciclo 1984

Published
2012

36. Cytokines and growth factors cross-link heparan sulfate

Author

Migliorini, Elisa, primary, Thakar, Dhruv, additional, Kühnle, Jens, additional, Sadir, Rabia, additional, Dyer, Douglas P., additional, Li, Yong, additional, Sun, Changye, additional, Volkman, Brian F., additional, Handel, Tracy M., additional, Coche-Guerente, Liliane, additional, Fernig, David G., additional, Lortat-Jacob, Hugues, additional, and Richter, Ralf P., additional

Published
2015
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46. Differential bioactivity of four BMP-family members as function of biomaterial stiffness

Author

Khodr, Paul Machillot, Guevara Garcia A, Laure Fourel, Albiges Rizo C, Sales Ramos A, Catherine Picart, Elisa Migliorini, Biomimétisme et Médecine Régénératrice (BRM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-BioSanté (UMR BioSanté), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Laboratoire des matériaux et du génie physique (LMGP ), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-Université Grenoble Alpes (UGA), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), ANR-17-CE13-0022,CODECIDE,Coincidence de detection dans l'identité cellulaire(2017), ANR-19-CE13-0031,GlyCON,Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique(2019), European Project: 259370,EC:FP7:ERC,ERC-2010-StG_20091028,BIOMIM(2011), European Project: 692924,H2020,ERC-2015-PoC,BIOACTIVECOATINGS(2016), Migliorini, Elisa, Coincidence de detection dans l'identité cellulaire - - CODECIDE2017 - ANR-17-CE13-0022 - AAPG2017 - VALID, Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique - Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique - - GlyCON2019 - ANR-19-CE13-0031 - AAPG2019 - VALID, Biomimetic films and membranes as advanced materials for studies on cellular processes - BIOMIM - - EC:FP7:ERC2011-06-01 - 2016-05-31 - 259370 - VALID, and BioActive Coatings in multi-well cell culture plates - BIOACTIVECOATINGS - - H20202016-04-01 - 2017-09-30 - 692924 - VALID

Subjects
animal structures, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Integrin, Biophysics, Bioengineering, Biocompatible Materials, Smad Proteins, SMAD, Article, Biomaterials, 03 medical and health sciences, 0302 clinical medicine, Transforming Growth Factor beta, Progenitor cell, Cell adhesion, Beta (finance), [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, 030304 developmental biology, 0303 health sciences, biology, Chemistry, 030302 biochemistry & molecular biology, Cell Differentiation, Bone Morphogenetic Protein Receptors, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Cell biology, [SDV.IB.BIO] Life Sciences [q-bio]/Bioengineering/Biomaterials, Mechanics of Materials, 030220 oncology & carcinogenesis, Bone Morphogenetic Proteins, embryonic structures, biology.protein, Ceramics and Composites, Phosphorylation, Alkaline phosphatase, Function (biology), Signal Transduction
Abstract

International audience; While a soft film itself is not able to induce cell spreading, BMP-2 presented via such soft film (so called“matrix-bound BMP-2”) was previously shown to trigger cell spreading, migration and downstream BMP-2signaling. Here, we used thin films of controlled stiffness presenting matrix-bound BMPs to study the effectof four BMP members (BMP-2, 4, 7, 9) on cell adhesion and differentiation of skeletal progenitors. Weperformed automated high content screening of cellular responses, including cell number, cell spreading area,SMAD phosphorylation and alkaline phosphatase activity. We revealed that the cell response to bBMPs isBMP-type specific and involved certain BMP receptors and beta chain integrins. In addition, this response isstiffness-dependent for several receptors. The basolateral presentation of the BMPs allowed us to discriminatethe specificity of cellular response and the role of type I and II BMP receptors and of β integrins in a BMPtypeand stiffness-dependent manner. Notably, the role of BMP-2 and BMP-4 were found to have distinctroles, while ALK5, previously known as a TGF-β receptor was revealed to be involved in the BMP-pathway.

Published
2022
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47. 3D-printed scaffold combined to 2D osteoinductive coatings to repair a critical-size mandibular bone defect

Author

Jean Boutonnat, Catherine Picart, Charlotte Garot, Sanela Morand, Georges Bettega, Vincent Fitzpatrick, Julien Vollaire, Paul Machillot, Véronique Josserand, M. Bouyer, Biomimétisme et Médecine Régénératrice (BRM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-BioSanté (UMR BioSanté), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Laboratoire des matériaux et du génie physique (LMGP ), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-Université Grenoble Alpes (UGA), Hôpital la Tronche, ANR-18-CE17-0016,OBOE,Régénération de défaut osseux induite par la surface d'un biomatériau(2018), European Project: 259370,EC:FP7:ERC,ERC-2010-StG_20091028,BIOMIM(2011), Migliorini, Elisa, APPEL À PROJETS GÉNÉRIQUE 2018 - Régénération de défaut osseux induite par la surface d'un biomatériau - - OBOE2018 - ANR-18-CE17-0016 - AAPG2018 - VALID, and Biomimetic films and membranes as advanced materials for studies on cellular processes - BIOMIM - - EC:FP7:ERC2011-06-01 - 2016-05-31 - 259370 - VALID

Subjects
Medicine (General), 3d printed, Scaffold, Materials science, QH301-705.5, Biomedical Engineering, Repair Kinetics, Bioengineering, 02 engineering and technology, Bone healing, Biomaterials, 03 medical and health sciences, chemistry.chemical_compound, R5-920, Polylactic acid, Tissue engineering, Full Length Article, Bone morphogenetic protein 2 (BMP-2), Biology (General), Bone regeneration, Molecular Biology, 030304 developmental biology, [SDV.IB] Life Sciences [q-bio]/Bioengineering, 0303 health sciences, 3D printing, Cell Biology, 021001 nanoscience & nanotechnology, Bone defect, Critical-size bone defect, 3. Good health, chemistry, [SDV.IB]Life Sciences [q-bio]/Bioengineering, 0210 nano-technology, Biotechnology, Biomedical engineering
Abstract

The reconstruction of large bone defects (12 cm3) remains a challenge for clinicians. We developed a new critical-size mandibular bone defect model on a minipig, close to human clinical issues. We analyzed the bone reconstruction obtained by a 3D-printed scaffold made of clinical-grade polylactic acid (PLA), coated with a polyelectrolyte film delivering an osteogenic bioactive molecule (BMP-2). We compared the results (computed tomography scans, microcomputed tomography scans, histology) to the gold standard solution, bone autograft. We demonstrated that the dose of BMP-2 delivered from the scaffold significantly influenced the amount of regenerated bone and the repair kinetics, with a clear BMP-2 dose-dependence. Bone was homogeneously formed inside the scaffold without ectopic bone formation. The bone repair was as good as for the bone autograft. The BMP-2 doses applied in our study were reduced 20- to 75-fold compared to the commercial collagen sponges used in the current clinical applications, without any adverse effects. Three-dimensional printed PLA scaffolds loaded with reduced doses of BMP-2 may be a safe and simple solution for large bone defects faced in the clinic., Graphical abstract Image 1, Highlights • We created a critical-size bone defect model in the mandible of mature minipigs, close to multiple clinical applications. • We engineered a new 3D-printed scaffold made of clinical-grade PLA and manufactured by FDM to fill the defect. • The BMP-2 dose released from the implants significantly influenced the amount of regenerated bone and the repair kinetics. • Bone reconstructed using this osteoinductive implant was homogeneous. • 3D-printed PLA scaffolds coated with 2D osteoinductive coatings lead to similar bone regeneration as bone autograft.

Published
2021
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48. Coarse-grained elastic network modelling: A fast and stable numerical tool to characterize mesenchymal stem cells subjected to AFM nanoindentation measurements

Author

Michele Gattullo, Michele Fiorentino, Elisa Migliorini, Elisabetta Ada Cavalcanti-Adam, Antonio Boccaccio, Vito Modesto Manghisi, Lorenzo Vaiani, A. E. Uva, Polytechnic University of Bari, Biomimétisme et Médecine Régénératrice (BRM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-BioSanté (UMR BioSanté), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Max Planck Institute for Medical Research [Heidelberg], Max-Planck-Gesellschaft, European Project: 658334,H2020,H2020-MSCA-IF-2014,OsteoNano(2015), Biomimetism and regenerative medicine (BRM), Migliorini, Elisa, and Spatial nanoscale control of growth and adhesion factors to enhance the osteogenic differentiation of mesenchymal stem cells - OsteoNano - - H20202015-05-01 - 2017-04-30 - 658334 - VALID

Subjects
Materials science, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [PHYS.MPHY]Physics [physics]/Mathematical Physics [math-ph], Modulus, Bioengineering, 02 engineering and technology, 010402 general chemistry, Microscopy, Atomic Force, 01 natural sciences, Quantitative Biology::Cell Behavior, Biomaterials, Mechanobiology, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Elastic Modulus, medicine, Meshfree methods, Humans, Computer Simulation, [MATH.MATH-RT] Mathematics [math]/Representation Theory [math.RT], Mechanical Phenomena, Cell Material Characterization, Microscopy, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [MATH.MATH-RT]Mathematics [math]/Representation Theory [math.RT], Atomic Force, Stiffness, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Elastic network modelAtomic force microscopyCell material characterizationMeshless methods, Mesenchymal Stem Cells, Nanoindentation, Atomic force microscopy, Cell material characterization, Elastic network model, Meshless methods, 021001 nanoscience & nanotechnology, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Finite element method, 0104 chemical sciences, Atomic Force Microscopy, Contact mechanics, Mechanics of Materials, medicine.symptom, 0210 nano-technology, Biological system, Material properties, Meshless Methods, Elastic Network Model
Abstract

International audience; The knowledge of the mechanical properties is the starting point to study the mechanobiology of mesenchymal stem cells and to understand the relationships linking biophysical stimuli to the cellular differentiation process. In experimental biology, Atomic Force Microscopy (AFM) is a common technique for measuring these mechanical properties. In this paper we present an alternative approach for extracting common mechanical parameters, such as the Young's modulus of cell components, starting from AFM nanoindentation measurements conducted on human mesenchymal stem cells. In a virtual environment, a geometrical model of a stem cell was converted in a highly deformable Coarse-Grained Elastic Network Model (CG-ENM) to reproduce the real AFM experiment and retrieve the related force-indentation curve. An ad-hoc optimization algorithm perturbed the local stiffness values of the springs, subdivided in several functional regions, until the computed force-indentation curve replicated the experimental one. After this curve matching, the extraction of global Young's moduli was performed for different stem cell samples. The algorithm was capable to distinguish the material properties of different subcellular components such as the cell cortex and the cytoskeleton. The numerical results predicted with the elastic network model were then compared to those obtained from hertzian contact theory and Finite Element Method (FEM) for the same case studies, showing an optimal agreement and a highly reduced computational cost. The proposed simulation flow seems to be an accurate, fast and stable method for understanding the mechanical behavior of soft biological materials, even for subcellular levels of detail. Moreover, the elastic network modelling allows shortening the computational times to approximately 33% of the time required by a traditional FEM simulation performed using elements with size comparable to that of springs.

Published
2021
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49. Additive Manufacturing of Material Scaffolds for Bone Regeneration: Toward Application in the Clinics

Author

Catherine Picart, Charlotte Garot, Georges Bettega, Biomimétisme et Médecine Régénératrice (BRM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-BioSanté (UMR BioSanté), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), ANR-18-CE17-0016,OBOE,Régénération de défaut osseux induite par la surface d'un biomatériau(2018), European Project: 259370,EC:FP7:ERC,ERC-2010-StG_20091028,BIOMIM(2011), Migliorini, Elisa, APPEL À PROJETS GÉNÉRIQUE 2018 - Régénération de défaut osseux induite par la surface d'un biomatériau - - OBOE2018 - ANR-18-CE17-0016 - AAPG2018 - VALID, and Biomimetic films and membranes as advanced materials for studies on cellular processes - BIOMIM - - EC:FP7:ERC2011-06-01 - 2016-05-31 - 259370 - VALID

Subjects
Pore size, [SDV.IB] Life Sciences [q-bio]/Bioengineering, Materials science, Material type, business.industry, 0206 medical engineering, Implantation Site, 3D printing, 02 engineering and technology, 021001 nanoscience & nanotechnology, Condensed Matter Physics, 020601 biomedical engineering, Article, Bone tissue engineering, Electronic, Optical and Magnetic Materials, Biomaterials, Tissue engineering, 3d image, Electrochemistry, [SDV.IB]Life Sciences [q-bio]/Bioengineering, 0210 nano-technology, Bone regeneration, business, ComputingMilieux_MISCELLANEOUS, Biomedical engineering
Abstract

Additive manufacturing (AM) allows the fabrication of customized bone scaffolds in terms of shape, pore size, material type and mechanical properties. Combined with the possibility to obtain a precise 3D image of the bone defects using computed tomography or magnetic resonance imaging, it is now possible to manufacture implants for patient-specific bone regeneration. This paper reviews the state-of-the-art of the different materials and AM techniques used for the fabrication of 3D-printed scaffolds in the field of bone tissue engineering. Their advantages and drawbacks are highlighted. For materials, specific criteria, were extracted from a literature study: biomimetism to native bone, mechanical properties, biodegradability, ability to be imaged (implantation and follow-up period), histological performances and sterilization process. AM techniques can be classified in three major categories: extrusion-based, powder-based and liquid-base. Their price, ease of use and space requirement are analyzed. Different combinations of materials/AM techniques appear to be the most relevant depending on the targeted clinical applications (implantation site, presence of mechanical constraints, temporary or permanent implant). Finally, some barriers impeding the translation to human clinics are identified, notably the sterilization process.

Published
2021
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50. Learning from BMPs and their biophysical extracellular matrix microenvironment for biomaterial design

Author

Corinne Albiges-Rizo, Elisa Migliorini, Amaris Guevara-Garcia, Catherine Picart, Biomimétisme et Médecine Régénératrice (BRM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-BioSanté (UMR BioSanté), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Laboratoire des matériaux et du génie physique (LMGP ), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-Université Grenoble Alpes (UGA), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), ANR-17-CE13-0022,CODECIDE,Coincidence de detection dans l'identité cellulaire(2017), ANR-19-CE13-0031,GlyCON,Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique(2019), European Project: 259370,EC:FP7:ERC,ERC-2010-StG_20091028,BIOMIM(2011), European Project: 334966,EC:FP7:ERC,ERC-2012-PoC,OSCODI(2013), European Project: 692924,H2020,ERC-2015-PoC,BIOACTIVECOATINGS(2016), Biomimetism and regenerative medicine (BRM), Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Universitaire [Grenoble] (CHU)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Migliorini, Elisa, Coincidence de detection dans l'identité cellulaire - - CODECIDE2017 - ANR-17-CE13-0022 - AAPG2017 - VALID, Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique - Décrypter la base biophysique par laquelle les glycosaminoglycanes contrôlent la signalisation des facteurs de croissance pendant le développement : une approche biomimétique - - GlyCON2019 - ANR-19-CE13-0031 - AAPG2019 - VALID, Biomimetic films and membranes as advanced materials for studies on cellular processes - BIOMIM - - EC:FP7:ERC2011-06-01 - 2016-05-31 - 259370 - VALID, Osteo-inductive coating of orthopedic and dental implants - OSCODI - - EC:FP7:ERC2013-07-01 - 2014-12-31 - 334966 - VALID, and BioActive Coatings in multi-well cell culture plates - BIOACTIVECOATINGS - - H20202016-04-01 - 2017-09-30 - 692924 - VALID

Subjects
0301 basic medicine, Histology, animal structures, Biomaterial design, Physiology, Endocrinology, Diabetes and Metabolism, media_common.quotation_subject, [SDV]Life Sciences [q-bio], BMPs, 030209 endocrinology & metabolism, Biocompatible Materials, [SDV.BC.IC] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Article, Extracellular matrix, 03 medical and health sciences, [SPI]Engineering Sciences [physics], 0302 clinical medicine, Bmp signaling, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Animals, Humans, Spatial localization, Mechanotransduction, Bone regeneration, Internalization, ComputingMilieux_MISCELLANEOUS, media_common, mechanotransduction, Extracellular Matrix Proteins, Chemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell biology, 030104 developmental biology, bioinspiration, Bone Morphogenetic Proteins, embryonic structures, Bioinspiration, Signal Transduction, biomaterials
Abstract

International audience; It is nowadays well-accepted that the extracellular matrix (ECM) is not a simple reservoir for growth factors but is an organization center of their biological activity. In this review, we focus on the ability of the ECM to regulate the biological activity of BMPs. In particular, we survey the role of the ECM components, notably the glycosaminoglycans and fibrillary ECM proteins, which can be promoters or repressors of the biological activities mediated by the BMPs. We examine how a process called mechano-transduction induced by the ECM can affect BMP signaling, including BMP internalization by the cells. We also focus on the spatio-temporal regulation of the BMPs, including their release from the ECM, which enables to modulate their spatial localization as well as their local concentration. We highlight how biomaterials can recapitulate some aspects of the BMPs/ECM interactions and help to answer fundamental questions to reveal previously unknown molecular mechanisms. Finally, the design of new biomaterials inspired by the ECM to better present BMPs is discussed, and their use for a more efficient bone regeneration in vivo is also highlighted.

Published
2020
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