Recently, nanosized cellulose materials extraction is extensively interesting from the sources of sustainable materials. Cellulose nano brils (CNF) extraction through green bio-based materials featured as promising interest in the eld of science. In this study, dimethyl sulfoxide (DMSO) was applied to examine its effectiveness in pretreating the Ficus natalensis barkcloth cellulose (FNBC) for CNF production perior to 2,2,6,6,-tetramethylpiperidine-1-oxyl (TEMPO) oxidation. The pretreatment performance of DMSO was evaluated based on the structural and morphological changes. DMSO pretreated FNBC attained the most dramatic morphological changes as compared to untreated cellulose samples. The results of scanning electron microscope (SEM) and transmission electron microscope (TEM) shows that there is an extentive structural disruption of FNBC during the pretreatment process, could be because of outstanding ability to eliminate non-cellulosic materials and amorphous regions from the FNBC, con rmed by the X-ray diffractometry (XRD) showing higher crystallinity values as well as higher thermal stablilites values of pretreated FNBC samples were also noted. Overall, this study revealed tremendously effective and pioneer pretreatment method for fractionating FNBC, to stimulate the successive extraction of cellulose nano brils. Furthermore, on the basis of the cellulose and CNF characterizations, this study showed that Ficus natalensis barkcloth could be considered as an alternative source of cellulose for potential value-added industrial applications such as food industry, paper making, and biomedicines.