Various cellulose nanofibrils (CNFs) created by refining and microfluidization, in combination with enzymatic or 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) oxidized pretreatment were compared. The morphological properties, degree of polymerization, and crystallinity for the obtained nanofibrils, as well as physical and mechanical properties of the corresponding films were evaluated. Compared to refining, intense microfluidization contributed greater separation of nanofibril bundles, which led to an enhancement of mechanical strength and transparency for the resultant film. The selected enzymatic pre-treatments produced shortened fibers due to preferential hydrolysis of amorphous cellulose and, in combination with mechanical treatments, resulted in short and stiff cellulose nanocrystal (CNC)-like materials. Despite films from these CNC-like fibrils having inferior tensile strength, their tensile modulus and transparency were significantly improved compared to CNFs prepared without pre-treatment. The unique fiber morphology and high crystallinity potentially offer a green and ecologically friendly alternative for the preparation of CNCs and CNFs as part of an integrated biorefinery approach. Published by Elsevier Ltd.