Control of Cell Surface and Functions by Layer-by-Layer Nanofilms

Abstract

Various nanometer-sized multilayers were directly prepared onto the surface of mouse L929 fibroblast cells by a layer-by-layer (LbL) assembly technique to control the cell surface microenvironment and cell functions, such as viability, morphology, and proliferation. The species of LbL nanofilms strongly affected the cell morphology and growth. Polyelectrolyte (PE) multilayers induced a round-shaped morphology of the adhered cells, although each component of the multilayers had high cytocompatibility, whereas fibronectin (FN)-gelatin (G) and -dextran sulfate (DS) multilayers with FN-binding domain interactions (FN films) showed extended morphologies of the cells similar to that of control cells (without films). A clear difference in cell proliferation was observed for PE and FN films. The cells with FN films on their surfaces showed good proliferation profiles independent of the film thickness, but cell proliferation was not observed using the PE films although the cells survived during the culture period. Fluorescence microscopic and scanning electron microscopic observations clearly suggested a nanometer-sized meshwork morphology of the FN films on the cell surface after 24 h of incubation, whereas the PE films showed homogeneous film morphologies on the cell surface. These nanomeshwork morphologies seemed to be similar to the fibrous structure of the natural extracellular matrix. The results of this study demonstrated that the components, charge, and morphology of LbL nanofilms prepared directly on the cell surface strongly affected cell functions, and the effects of these LbL nanofilms on cell functions differed vastly as compared to PE films prepared on a substrate. The preparation of LbL nanofilms onto a cell surface might be a novel and interesting technique to control cell functions.

Development of Three-Dimensional Tissue Models Based on Hierarchical Cell Manipulation Using Nanofilms

“…The nanometer-sized meshwork morphology of the FN films on the cell surface was found after 24 h of incubation, whereas the PE films showed homogeneous film morphologies on the cell surface (Figures 5h5j). 26 These nanomeshwork morphologies seemed to be similar to the fibrous structure of the natural ECM.…”

“…These results suggested that 4L-NHDF provided a more favorable environment for HUVECs than a cell culture plastic disk to induce high thermosensitivity and to suppress the inflammatory response from the substrate. Since the FNG nanofilms prepared on the cell surface did not affect cellular functions, 26 we concluded that the layered construct would be an analogous environment to natural tissues. These findings could be important for not only tissue engineering, but also for basic cell biology.…”

“…2325 Accordingly, various LbL films consisting of synthetic polymers, polysaccharides, poly(amino acid)s, and proteins were prepared onto the surface of mouse L929 fibroblasts, and the cell viability, morphology, and proliferation were investigated in relation to the LbL films, because we would like to evaluate the effect of LbL films on basic cellular functions to clarify the universal effect of the LbL films prepared on the cell surface. 26 Step Number with the 7-step-assembled, (e, f ) without a 7-step-assembled, or (g, h) with a 1-step-assembled FNG nanofilms on the surface of the fist cell layer. i) Ph image of a L929 cell monolayer as a control.…”

Rapid Construction of Three‐Dimensional Multilayered Tissues with Endothelial Tube Networks by the Cell‐Accumulation Technique

“…The FN-G nanofi lms, negatively charged under physiological conditions, were prepared by interaction through the collagen-binding domain of FN without a cytotoxic cationic component, and can provide high cyto-compatibility like native ECM. [ 21 ] The adhesion and proliferation properties of cells coated with the FN-G fi lms were comparable to non-coated cells ( Figure S3). We tried to perform the rapid construction of multilayered tissues using hFCs coated with FN-G nanofi lms.…”

Three-Dimensional Tissue Models Constructed by Cells with Nanometer- or Micrometer-Sized Films on the Surfaces

“…FN and DS had negative charges under physiological conditions. [55] Furthermore, we also successfully applied polyelectrolyte (PE) films, for example, FN-E-Lys and E-Lys-PSS, on the cell surfaces based on their common electrostatic interactions between the positive and negative components. To evaluate the LbL nanometer-sized film coating effects on cell functions, mouse L929 cells with and without different kinds of LbL films were used for proliferation experiments.…”