Lignocellulosic resources have been recognized to be the most promising sustainable and renewable source of materials for the elaboration of composites (Thakur et al, 2014). Notably, plant cell walls have are multilayered materials resulting from the assembly of different biopolymers: cellulose, hemicelluloses and lignins. These components are interconnected through a variety of covalent and non-covalent bonds that make a highly organized network. This structural organization plays a key role in the regulation of plant cell growth, mechanical properties and biodegradation. Chemical and structural variations within cell wall layers will not only affect the mechanical properties of the fibers but also the different preparation steps such as the defibration, and the interface properties between the fibers and the matrix in the composites (Salmen and Burgert, 2009, Burgert and Keplinger, 2013). Therefore, anticipating and controlling the mechanical properties of the composites implies to evaluate a multi-scale analysis of the morphological and physicochemical properties of the fibers (Le Digou et al, 2014). Nanoscale investigation of the chemical and physical properties of the cell wall could provide a more comprehensive view on the molecular and supramolecular mechanisms that promote their physicochemical properties and reactivity is still challenging In order to get information of the fiber properties at nanoscale level, the potentialities of atomic force microscopy (AFM) were assessed in different modes while comparing plant cell walls and of bioinspired polymer films. To this end, non-lignified and lignified systems were obtained using cellulose nanocrystals, hemicelluloses and lignin as single, binary and ternary nanocomposites (Muraille et al, 2015). Thanks to the use of adhesion measurements via tip functionalization with lignocellulosic polymers (Estephan et al, 2011), of nanoInfrared absorption, nanomechanical measurements, and by comparison between native cell walls (poplar, hemp) and lignocellulosic bioinspired films, we will try to understand the relationship between the composition and the supramolecular organization of lignocellulosic polymers and their nanoscale properties.