Membrane Surface Nanostructures and Adhesion Property of T Lymphocytes Exploited by AFM

The activation of T lymphocytes plays a very important role in T-cell-mediated immune response. Though there are many related literatures, the changes of membrane surface nanostructures and adhesion property of T lymphocytes at different activation stages have not been reported yet. However, these investigations will help us further understand the biophysical and immunologic function of T lymphocytes in the context of activation. In the present study, the membrane architectures of peripheral blood T lymphocytes were obtained by AFM, and adhesion force of the cell membrane were measured by acquiring force–distance curves. The results indicated that the cell volume increased with the increases of activation time, whereas membrane surface adhesion force decreased, even though the local stiffness for resting and activated cells is similar. The results provided complementary and important data to further understand the variation of biophysical properties of T lymphocytes in the context of in vitro activation

Force and Compliance Measurements on Living Cells Using Atomic Force Microscopy (AFM)

We describe the use of atomic force microscopy (AFM) in studies of cell adhesion and cell compliance. Our studies use the interaction between leukocyte function associated antigen-1 (LFA-1)/intercellular adhesion molecule-1 (ICAM-1) as a model system. The forces required to unbind a single LFA-1/ICAM-1 bond were measured at different loading rates. This data was used to determine the dynamic strength of the LFA-1/ICAM-1 complex and characterize the activation potential that this complex overcomes during its breakage. Force measurements acquired at the multiple- bond level provided insight about the mechanism of cell adhesion. In addition, the AFM was used as a microindenter to determine the mechanical properties of cells. The applications of these methods are described using data from a previous study

Nano-surgery at the leukocyte–endothelial docking site

The endothelium has an important role in controlling the extravasation of leukocytes from blood to tissues. Endothelial permeability for leukocytes is influenced by transmembrane proteins that control inter-endothelial adhesion, as well as steps of the leukocyte transmigration process. In a cascade consisting of leukocyte rolling, adhesion, firm adhesion, and diapedesis, a new step was recently introduced, the formation of a docking structure or “transmigratory cup.” Both terms describe a structure formed by endothelial pseudopods embracing the leukocyte. It has been found associated with both para- and transcellular diapedesis. The aim of this study was to characterize the leukocyte–endothelial contact area in terms of morphology and cell mechanics to investigate how the endothelial cytoskeleton reorganizes to engulf the leukocyte. We used atomic force microscopy (AFM) to selectively remove the leukocyte and then analyze the underlying cell at this specific spot. Firmly attached leukocytes could be removed by AFM nanomanipulation. In few cases, this exposed 8–12 μm wide and 1 μm deep footprints, representing the cup-like docking structure. Some of them were located near endothelial cell junctions. The interaction area did not exhibit significant alterations neither morphologically nor mechanically as compared to the surrounding cell surface. In conclusion, the endothelial invagination is formed without a net depolymerization of f-actin, as endothelial softening at the site of adhesion does not seem to be involved. Moreover, there were no cases of phagocytotic engulfment, but instead the formation of a transmigratory channel could be observed

Surface functionalization of titanium with silver nanoparticles

This study aims to investigate the most efficient ways for metallic samples functionalization with silver nanoparticles (AgNPs). Three different techniques of surface functionalization have been used for the coating of titanium metal, i.e. the sessile drop method (evaporation), dip-coating and electrophoretic deposition (EPD). AgNPs stabilized with polyvinylpyrrolidone had a spherical shape and the metallic core diameter, charge and polydispersity index were 70±20 nm, -15 mV and 0.192, respectively. SEM analysis revealed that AgNPs were homogeneously distributed over the entire surface and did not form the particle agglomerates only in case of EPD. Thus, EPD method and spherical AgNPs can be used for further investigation concerning the preparation of biocomposites with antibacterial and bioactive properties

Quantifying molecular-level cell adhesion on electroactive conducting polymers using electrochemical-single cell force spectroscopy

Single Cell Force Spectroscopy was combined with Electrochemical-AFM to quantify the adhesion between live single cells and conducting polymers whilst simultaneously applying a voltage to electrically switch the polymer from oxidized to reduced states. The cell-conducting polymer adhesion represents the non-specific interaction between cell surface glycocalyx molecules and polymer groups such as sulfonate and dodecylbenzene groups, which rearrange their orientation during electrical switching. Single cell adhesion significantly increases as the polymer is switched from an oxidized to fully reduced state, indicating stronger cell binding to sulfonate groups as opposed to hydrophobic groups. This increase in single cell adhesion is concomitant with an increase in surface hydrophilicity and uptake of cell media, driven by cation movement, into the polymer film during electrochemical reduction. Binding forces between the glycocalyx and polymer surface are indicative of molecular-level interactions and during electrical stimulation there is a decrease in both the binding force and stiffness of the adhesive bonds. The study provides insight into the effects of electrochemical switching on cell adhesion at the cell-conducting polymer interface and is more broadly applicable to elucidating the binding of cell adhesion molecules in the presence of electrical fields and directly at electrode interfaces