Polymer gel-tethered lipid bilayers (PGTB) represent attractive cell surface-mimicking substrates for the study of cellular mechanosensitivity. A hallmark of these polymer-lipid composites is that the properties of their polymer component and the nature of the polymer-lipid linkages may have a profound impact on the organization and dynamics of the polymer gel-attached lipid bilayer. To better understand the role of the polymer gel attachment on lipid bilayer properties, here we explore the membrane organization and lipid dynamics in two complementary PGTB architectures. In the first case, a PGTB is assembled in a sandwiched architecture between two glass slides by bringing together a glass-supported lipid monolayer with another lipid monolayer, which is attached to the surface of functionalized polyacrylamide (PAA) gel layer via stable maleimide-thiol linkages. In the second case, a PGTB is built in an open geometry, which would be suitable as artificial cell substrate. Here, a lipid monolayer is first attached to the surface of a functionalized PAA gel followed by addition of giant unilamellar vesicles (GUV), which are caused to rupture and fuse into a PGTB via osmotic stress. The membrane organization and dynamics of both PGTB architectures is analyzed using epifluorescence microscopy, fluorescence recovery after photobleaching (FRAP), and confocal fluorescence correlation spectroscopy (FCS).

Membrane organization and dynamics in polymer gel-tethered lipid bilayer (PGTB) systems

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