10:30 AM - SB02.07.09
Cell-Biomaterial Interface Visualized at the Nanoscale
Francesca Santoro1,Laura Matino1,Paola Scognamiglio1,Valentina Mollo1
Istituto Italiano di Tecnologia1
Cell instructive biomaterials interact directly with biological systems, modifying their behavior by fine-tuning the crosstalk at cellular level. Cell spreading, polarization and cellular mechanical tension are only few examples of interface dynamics consequences. Great consideration has been given to the role of the cell membrane and the surface tension induced by the topography of the material in contact with it. In this context, many efforts have been focused in the last decade to characterize the cell-membrane interface, at the relevant scale. Beside the standard optical acquisition that allows imaging with major limitations in resolution (~ 100 nm), electron microscopy based-acquisition exceeds that limit. However, high resolution electron microscopy procedures require long specimen preparation processes as well as the imaging and processing of hundreds of specimen sections. Here, we present an advanced microscopy method (scanning electron microscopy/focused ion beam) based on ultra-thin resin plastificization which uniquely allows the visualization of the interface between cells and materials with 5-10 nm resolution (1,2,3). This technique allows for the visualization of a region of interest where the cell is in contact with the biomaterial underneath. The use of focused ion beam allows for etching through a variety of materials. In fact, here we will present relevant adhesion process of cells in contact with organic and inorganic materials, pseudo 3D materials (vertical nanostructures) and 3D scaffolds. Our results could give new insights in designing new efficient 3D structures for tissue engineering purpose and their interaction and effect on cellular ultrastructures at the cell-biomaterial interface.
(1) Li X., Matino L., Zhang W., Klausen L., McGuire A.F., Lubrano C., Zhao W., Santoro F., Cui B., A nanostructure plaform for live-cell manipulation of membrane curvature. Nature Protocols, 2019
(2) Santoro, F., Zhao, W., Joubert, L.-M., Duan, L., Schnitker, J., van de Burgt, Y., Lou, H.-Y., Liu, B., Salleo, A., Cui, L., Cui Y., Cui B., Revealing the Cell–Material Interface with Nanometer Resolution by Focused Ion Beam/Scanning Electron Microscopy, ASC Nano, 2017.
(3) Zhao, W., Hanson, L., Lou, H.-Y., Akamatsu, M., Chowdary, P.D., Santoro, F., Marks, J.R., Grassart, A., Drubin, D.G., Cui, Y., Cui B., Nanoscale manipulation of membrane curvature for probing endocytosis in live cells. Nature Nanotechnology, 2017.
(4) Iandolo D., Pennacchio F. A., Mollo V., Rossi D., Dannhauser D., Cui B., Owens R. M., Santoro F., Electron Microscopy for 3D Scaffolds–Cell Biointerface Characterization, Advanced Biosystems, 2018.