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7. Nanomechanical and topographical imaging of living cells by Atomic Force Microscopy with colloidal probes

Author

Puricelli, Luca, Galluzzi, Massimiliano, Schulte, Carsten, Podestà, Alessandro, and Milani, Paolo

Subjects
Condensed Matter - Soft Condensed Matter, Physics - Biological Physics, Quantitative Biology - Cell Behavior
Abstract

Atomic Force Microscopy (AFM) has a great potential as a tool to characterize mechanical and morphological properties of living cells; these properties have been shown to correlate with cells' fate and patho-physiological state in view of the development of novel early-diagnostic strategies. Although several reports have described experimental and technical approaches for the characterization of cell elasticity by means of AFM, a robust and commonly accepted methodology is still lacking. Here we show that micrometric spherical probes (also known as colloidal probes) are well suited for performing a combined topographic and mechanical analysis of living cells, with spatial resolution suitable for a complete and accurate mapping of cell morphological and elastic properties, and superior reliability and accuracy in the mechanical measurements with respect to conventional and widely used sharp AFM tips. We address a number of issues concerning the nanomechanical analysis, including the applicability of contact mechanical models and the impact of a constrained contact geometry on the measured elastic modulus (the finite-thickness effect). We have tested our protocol by imaging living PC12 and MDA-MB-231 cells, in order to demonstrate the importance of the correction of the finite-thickness effect and the change in cell elasticity induced by the action of a cytoskeleton-targeting drug., Comment: 51 pages, 12 figures, 3 tables

Published
2014
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11. Correction: Niu, Y.; Galluzzi, M. Hyaluronic Acid/Collagen Nanofiber Tubular Scaffolds Support Endothelial Cell Proliferation, Phenotypic Shape and Endothelialization. Nanomaterials 2021, 11 , 2334.

Author

Niu, Yuqing and Galluzzi, Massimiliano

Subjects
*METAL scaffolding, *FLUORESCENCE microscopy, *CONTACT angle, *HYALURONIC acid, *MANNEQUINS (Figures)
Abstract

This document is a correction notice for an article titled "Hyaluronic Acid/Collagen Nanofiber Tubular Scaffolds Support Endothelial Cell Proliferation, Phenotypic Shape and Endothelialization" published in the journal Nanomaterials. The correction addresses errors in Figures 1B, 2A, and 5A of the original publication. The corrected figures are provided in the document. The authors apologize for any inconvenience caused but state that the scientific conclusions of the article remain unaffected. [Extracted from the article]

Published
2024
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12. Polydopamine-Mediated Protein Adsorption Alters the Epigenetic Status and Differentiation of Primary Human Adipose-Derived Stem Cells (hASCs)

Author

Harati, Javad, primary, Tao, Xuelian, additional, Shahsavarani, Hosein, additional, Du, Ping, additional, Galluzzi, Massimiliano, additional, Liu, Kun, additional, Zhang, Zhen, additional, Shaw, Peter, additional, Shokrgozar, Mohammad Ali, additional, Pan, Haobo, additional, and Wang, Peng-Yuan, additional

Published
2022
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20. Additional file 1 of In vivo performance of electrospun tubular hyaluronic acid/collagen nanofibrous scaffolds for vascular reconstruction in the rabbit model

Author

Niu, Yuqing, Galluzzi, Massimiliano, Fu, Ming, Hu, Jinhua, and Xia, Huimin

Abstract

Additional file 1: Figure S1. Vascular grafts fabricated with cellularized tubular HA/collagen nanofibrous scaffold facilitates vascular reconstruction. H&E staining the cross-section of a retrieved tubular HA/collagen nanofibrous grafts 6 weeks after transplant at 2.5 $$\times $$ �� (upper panel) and 40 $$\times $$ �� (lower panel) compared with rabbit carotid artery (b, d, g). Scale bars, 100 ��m. Table S1. The size of the obtained electrospun tubular HA/collagen nanofibrous scaffolds.

Published
2021
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21. A biomimetic hyaluronic acid‐silk fibroin nanofiber scaffold promoting regeneration of transected urothelium.

Author

Niu, Yuqing, Galluzzi, Massimiliano, Deng, Fuming, Zhao, Zhang, Fu, Ming, Su, Liang, Sun, Weitang, Jia, Wei, and Xia, Huimin

Subjects
*UROTHELIUM, *REGENERATION (Biology), *URINARY organs, *SILK fibroin, *HYALURONIC acid, *POLYCAPROLACTONE, *BONE regeneration, KERATINOCYTE differentiation
Abstract

This study was designed to investigate the regulatory effect of hyaluronic acid (HA)—coating silk fibroin (SF) nanofibers during epithelialization of urinary tract for urethral regeneration. The obtained electrospun biomimetic tubular HA‐SF nanofiber scaffold is composed of a dense inner layer and a porous outer layer in order to mimic adhesion and cavernous layers of the native tissue, respectively. A thin layer of HA‐gel coating was fixed in the inner wall to provide SF nanofibers with a dense and smooth surface nano‐topography and higher hydrophilicity. Compared with pure SF nanofibers, HA‐SF nanofibers significantly promoted the adhesion, growth, and proliferation of primary urothelial cells, and up‐regulate the expression of uroplakin‐3 (terminal differentiation keratin protein in urothelium). Using the New Zealand male rabbit urethral injury model, the scaffold composed of tubular HA‐SF nanofibers could recruit lumen and myoepithelial cells from the adjacent area of the host, rapidly reconstructing the urothelial barrier in the wound area in order to keep the urinary tract unobstructed, thereby promoting luminal epithelialization, smooth muscle bundle structural remodeling, and capillary formation. Overall, the synergistic effects of nano‐topography and biophysical cues in a biomimetic scaffold design for effective endogenous regeneration. [ABSTRACT FROM AUTHOR]

Published
2022
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25. Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples

Author

Schillers, Hermann, Rianna, Carmela, Schäpe, Jens, Luque, Tomas, Doschke, Holger, Wälte, Mike, Uriarte, Juan José, Campillo, Noelia, Michanetzis, Georgios P. A., Bobrowska, Justyna, Dumitru, Andra, Herruzo, Elena T., Bovio, Simone, Parot, Pierre, Galluzzi, Massimiliano, Podestà, Alessandro, Puricelli, Luca, Scheuring, Simon, Missirlis, Yannis, Garcia, Ricardo, Odorico, Michael, Teulon, Jean-Marie, Lafont, Frank, Lekka, Malgorzata, Rico, Felix, Rigato, Annafrancesca, Pellequer, Jean-Luc, Oberleithner, Hans, Navajas, Daniel, Radmacher, Manfred, Westfälische Wilhelms-Universität Münster ( WWU ), University of Bremen, University of Barcelona, University of Patras [Patras], Polish Academy of Sciences, Instituto de Ciencia de Materiales de Madrid [Madrid] ( ICMM ), Institut Pasteur de Lille, Réseau International des Instituts Pasteur ( RIIP ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Università degli studi di Milano [Milano], Department Radiation Oncology (Weill Cornell Medicine), Weill Medical College of Cornell University [New York], Institut de Chimie Séparative de Marcoule ( ICSM - UMR 5257 ), Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Ecole Nationale Supérieure de Chimie de Montpellier ( ENSCM ) -Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de biologie structurale ( IBS - UMR 5075 ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ), BIO-AFM-LAB Bio Atomic Force Microscopy Laboratory ( Bio-AFM-Lab ), Aix Marseille Université ( AMU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), ANR-10-EQPX-0004/10-EQPX-0004,Imaginex BioMed,Plateau de microscopie de criblage à haut débit et d'analyse à très haute résolution ( 2010 ), European Project : 340177,EC:FP7:ERC,ERC-2013-ADG,3DNANOMECH ( 2014 ), Westfälische Wilhelms-Universität Münster (WWU), Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Réseau International des Instituts Pasteur (RIIP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Università degli Studi di Milano [Milano] (UNIMI), Etude de la Matière en Mode Environnemental (L2ME), Institut de Chimie Séparative de Marcoule (ICSM - UMR 5257), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-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)-Centre National de la Recherche Scientifique (CNRS), BIO-AFM-LAB Bio Atomic Force Microscopy Laboratory (Bio-AFM-Lab), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), ANR-10-EQPX-0004,Imaginex BioMed,Plateau de microscopie de criblage à haut débit et d'analyse à très haute résolution(2010), European Project: 340177,EC:FP7:ERC,ERC-2013-ADG,3DNANOMECH(2014), Universitat de Barcelona, Ministerio de Ciencia e Innovación (España), International AFMBioMed Conferences Association, Università degli Studi di Milano, Ministerio de Economía y Competitividad (España), German Research Foundation, European Research Council, Agence Nationale de la Recherche (France), National Science Centre (Poland), Westfälische Wilhelms-Universität Münster = University of Münster (WWU), University of Patras, Polska Akademia Nauk = Polish Academy of Sciences (PAN), Università degli Studi di Milano = University of Milan (UNIMI), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Nanotecnologia, Science, technology, industry, and agriculture, Reproducibility of Results, Biomecànica, Hydrogels, Microscòpia de força atòmica, Microscopy, Atomic Force, Article, Madin Darby Canine Kidney Cells, Atomic force microscopy, Dogs, Elastic Modulus, Medicine, Animals, Nanotechnology, Biomechanics, Stress, Mechanical, [ SDV.BBM.BS ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

We present a procedure that allows a reliable determination of the elastic (Young's) modulus of soft samples, including living cells, by atomic force microscopy (AFM). The standardized nanomechanical AFM procedure (SNAP) ensures the precise adjustment of the AFM optical lever system, a prerequisite for all kinds of force spectroscopy methods, to obtain reliable values independent of the instrument, laboratory and operator. Measurements of soft hydrogel samples with a well-defined elastic modulus using different AFMs revealed that the uncertainties in the determination of the deflection sensitivity and subsequently cantilever's spring constant were the main sources of error. SNAP eliminates those errors by calculating the correct deflection sensitivity based on spring constants determined with a vibrometer. The procedure was validated within a large network of European laboratories by measuring the elastic properties of gels and living cells, showing that its application reduces the variability in elastic moduli of hydrogels down to 1%, and increased the consistency of living cells elasticity measurements by a factor of two. The high reproducibility of elasticity measurements provided by SNAP could improve significantly the applicability of cell mechanics as a quantitative marker to discriminate between cell types and conditions., This work was supported by grants to D.N., T.L., N.C., and J.J.U. (Spanish Ministry of Economy and Competitiveness FIS-PI14/00280), M.L. and J.B. (National Science Center NCN DEC-2011/01/M/ST3/00711), H.O. (Deutsche Forschungsgemeinschaft, Koselleck 68/1 and Centre of Excellence, Cells in Motion (CIM), University of Münster, Germany), S.B. and F.L. (ANR-10-EQPX-04-01) and A.D., E.T.H., R.G. (CSD2010-00024, ERC-AdG-340177). A.P. thanks the Dept. of Physics of the University of Milano for financial support under the project “Piano di Sviluppo dell’Ateneo per la Ricerca 2014”, actions: “A. Upgrade of instrumentation”, and “B. Support to young researchers”. We acknowledge the support of the International AFMBioMed Conference Association.

Published
2017

27. Standardized nanomechanical atomic force microscopy procedure (SNAP) for measuring soft and biological samples

Author

Ministerio de Ciencia e Innovación (España), International AFMBioMed Conferences Association, Università degli Studi di Milano, Ministerio de Economía y Competitividad (España), German Research Foundation, European Research Council, Agence Nationale de la Recherche (France), National Science Centre (Poland), Schillers, Hermann, Rianna, Carmela, Schäpe, Jens, Luque, Tomás, Doschke, Holger, Wälte, Mike, Uriarte, Juan José, Campillo, Noelia, Michanetzis, Georgios P. A., Bobrowska, Justyna, Dumitru, Andrea, Herruzo, Elena T., Bovio, Simone, Parot, Pierre, Galluzzi, Massimiliano, Podestà, Alessandro, Puricelli, Luca, Scheuring, Simon, Missirlis, Yannis, García García, Ricardo, Odorico, Michael, Teulon, Jean-Marie, Lafont, Frank, Lekka, Malgorzata, Rico, Felix, Rigato, Annafrancesca, Pellequer, Jean-Luc, Oberleithner, Hans, Navajas, Daniel, Radmacher, Manfred, Ministerio de Ciencia e Innovación (España), International AFMBioMed Conferences Association, Università degli Studi di Milano, Ministerio de Economía y Competitividad (España), German Research Foundation, European Research Council, Agence Nationale de la Recherche (France), National Science Centre (Poland), Schillers, Hermann, Rianna, Carmela, Schäpe, Jens, Luque, Tomás, Doschke, Holger, Wälte, Mike, Uriarte, Juan José, Campillo, Noelia, Michanetzis, Georgios P. A., Bobrowska, Justyna, Dumitru, Andrea, Herruzo, Elena T., Bovio, Simone, Parot, Pierre, Galluzzi, Massimiliano, Podestà, Alessandro, Puricelli, Luca, Scheuring, Simon, Missirlis, Yannis, García García, Ricardo, Odorico, Michael, Teulon, Jean-Marie, Lafont, Frank, Lekka, Malgorzata, Rico, Felix, Rigato, Annafrancesca, Pellequer, Jean-Luc, Oberleithner, Hans, Navajas, Daniel, and Radmacher, Manfred

Abstract

We present a procedure that allows a reliable determination of the elastic (Young's) modulus of soft samples, including living cells, by atomic force microscopy (AFM). The standardized nanomechanical AFM procedure (SNAP) ensures the precise adjustment of the AFM optical lever system, a prerequisite for all kinds of force spectroscopy methods, to obtain reliable values independent of the instrument, laboratory and operator. Measurements of soft hydrogel samples with a well-defined elastic modulus using different AFMs revealed that the uncertainties in the determination of the deflection sensitivity and subsequently cantilever's spring constant were the main sources of error. SNAP eliminates those errors by calculating the correct deflection sensitivity based on spring constants determined with a vibrometer. The procedure was validated within a large network of European laboratories by measuring the elastic properties of gels and living cells, showing that its application reduces the variability in elastic moduli of hydrogels down to 1%, and increased the consistency of living cells elasticity measurements by a factor of two. The high reproducibility of elasticity measurements provided by SNAP could improve significantly the applicability of cell mechanics as a quantitative marker to discriminate between cell types and conditions.

Published
2017

31. Conversion of nanoscale topographical information of cluster-assembled zirconia surfaces into mechanotransductive events promotes neuronal differentiation

Author

Schulte, Carsten, primary, Rodighiero, Simona, additional, Cappelluti, Martino Alfredo, additional, Puricelli, Luca, additional, Maffioli, Elisa, additional, Borghi, Francesca, additional, Negri, Armando, additional, Sogne, Elisa, additional, Galluzzi, Massimiliano, additional, Piazzoni, Claudio, additional, Tamplenizza, Margherita, additional, Podestà, Alessandro, additional, Tedeschi, Gabriella, additional, Lenardi, Cristina, additional, and Milani, Paolo, additional

Published
2016
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33. Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples.

Author

Schillers H, Rianna C, Schäpe J, Luque T, Doschke H, Wälte M, Uriarte JJ, Campillo N, Michanetzis GPA, Bobrowska J, Dumitru A, Herruzo ET, Bovio S, Parot P, Galluzzi M, Podestà A, Puricelli L, Scheuring S, Missirlis Y, Garcia R, Odorico M, Teulon JM, Lafont F, Lekka M, Rico F, Rigato A, Pellequer JL, Oberleithner H, Navajas D, and Radmacher M

Subjects
Animals, Dogs, Elastic Modulus, Madin Darby Canine Kidney Cells, Nanotechnology, Reproducibility of Results, Stress, Mechanical, Hydrogels chemistry, Microscopy, Atomic Force methods
Abstract

We present a procedure that allows a reliable determination of the elastic (Young's) modulus of soft samples, including living cells, by atomic force microscopy (AFM). The standardized nanomechanical AFM procedure (SNAP) ensures the precise adjustment of the AFM optical lever system, a prerequisite for all kinds of force spectroscopy methods, to obtain reliable values independent of the instrument, laboratory and operator. Measurements of soft hydrogel samples with a well-defined elastic modulus using different AFMs revealed that the uncertainties in the determination of the deflection sensitivity and subsequently cantilever's spring constant were the main sources of error. SNAP eliminates those errors by calculating the correct deflection sensitivity based on spring constants determined with a vibrometer. The procedure was validated within a large network of European laboratories by measuring the elastic properties of gels and living cells, showing that its application reduces the variability in elastic moduli of hydrogels down to 1%, and increased the consistency of living cells elasticity measurements by a factor of two. The high reproducibility of elasticity measurements provided by SNAP could improve significantly the applicability of cell mechanics as a quantitative marker to discriminate between cell types and conditions.

Published
2017
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