This study aimed to produce, purify, and characterize SCOBY nanocellulose for biomedical use. The material was obtained through a biotechnological fermentation process of tea broth using a symbiotic culture of bacteria and yeast (SCOBY). The tea broth consisted of black tea extract supplemented with sucrose. The SCOBY pellicle formed at the air-liquid interface after 10 days of fermentation was collected. Pellicle purification involved washing with Milli-Q water, sonication combined with ultraviolet-C (UV-C) treatment, followed by washing with hot 1.5 M NaOH and 1.5% H₂O₂ solutions. Fourier transform infrared (FTIR) spectroscopy confirmed the effectiveness of the purification process, leading to the acquisition of a transparent nanocellulose hydrogel. Scanning electron microscopy (SEM) revealed the characteristic nanofibrous morphology of SCOBY nanocellulose, with nanofibers exhibiting a mean diameter of 88 ± 19 nm. The nanofibers formed a highly porous internal structure, whereas the outer layer consisted predominantly of densely packed nanofibers, effectively protecting the culture from the external environment. The pores within the structure exhibited diameters ranging from 4 to 6 µm. The mean density of the structure was 0.024 ± 0.006 g/cm³. In vitro studies using L929 murine fibroblasts demonstrated that SCOBY nanocellulose was not cytotoxic, as confirmed by the Alamar Blue assay and live-dead fluorescence staining.