β-Cyclodextrin (β-CD)/cellulose hydrogels were prepared in NaOH/urea aqueous solution by crosslinking with epichlorohydrin. The structure and morphology of the hydrogels were characterized with Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM). The swelling test, 5-fluorouracil (5-FU) and bovine serum albumin (BSA), and aniline blue (AnB) were used to investigate the swelling capability, drug release behavior and the fluorescent property of the hydrogels. The results indicated that the swelling degree and water uptake of the hydrogels decreased with an increase of the β-CD content. The in vitro release of 5-FU and BSA of the hydrogels showed an inclusion complex formed between 5-FU and β-CD. β-CD/cellulose hydrogels adsorbed AnB lead to a fluorescence enhancement attributing to the formation of the host-guest complex between β-CD and AnB.