The glycocalyx affects force loading-dependent mechanotransductive topography sensing at the nanoscale

The cell/microenvironment interface is the starting point of integrin-mediated mechanotransduction, but many details of mechanotransductive signal integration remain elusive due to the complexity of the involved (extra)cellular structures, such as the glycocalyx. We used engineered nano-bio interfaces with extracellular matrix nanotopography-mimicking features to analyse the impact of the glycocalyx on nano-mechanosensing. Our data demonstrates that the glycocalyx configuration affects spatiotemporal nanotopography-sensitive mechanotransductive events at the cell/microenvironment interface. Opposing effects of glycocalyx removal were observed, when comparing flat and specific nanotopographical conditions (i.e., 15 nm root-mean-square (rms) roughness). In fact, the excessive force loading and retrograde actin flow speed, characteristic for the 15 nm rms nanotopography in the presence of native glycocalyx, are strongly reduced in its absence. Conversely, on the flat substrate, these parameters increased upon glycocalyx-targeting enzymatic treatment. Our results highlight the importance of the glycocalyx configuration in a molecular clutch force loading-dependent cellular mechanism for nano-mechanosensing of the topography.