Biorefinery aims at designing new virtuous and high-efficiency energy chains, achieving the combined production of biofuels (e.g., bioethanol) and biobased products. This emerging philosophy can represent an important opportunity for the industrial world, exploiting a new kind of nano-smart biomaterials in their production chains. This paper will present the lab experience carried out by the Biomass Research Centre (CRB) in extracting cellulose nanocrystals (NCC) from a pretreated (via Steam Explosion) fraction of Cynara cardunculus. This is a very common and invasive arboreal variety in central Italy. The NCC extraction methodology allows the separation of the crystalline content of cellulose. Such a procedure has been considered in the literature with the exception of one step in which the conditions have been optimized by CRB Lab. This procedure has been applied for the production of NCC from both Cynara cardunculus and microcrystalline cellulose (MCC). The paper will discuss some of the results achieved using the obtained nanocrystals as reinforcing filler in a paper sheet; it was found that the tensile strength increased from 3.69 kg/15 mm to 3.98 kg/15 mm, the durability behavior (measured by bending number) changed from the value 95 to the value 141, and the barrier properties (measured by Gurley porosity) were improved, increasing from 38 s to 45 s.