Fullerene is a spherical hollow structure composed of carbon atoms. It has emerged in the new development of numerous fullerene-based compounds with a variety of biological targets such as anticancer, cytoprotection, tissue regeneration, and controlled drug delivery. Because of its biodegradable property of chitosan hydrogels, fullerine can function as a drug delivery system for implants with optimal degradation profiles that result in the proper rates of drug release. In this study, a chitosan-fullerene hybrid hydrogel was fabricated, and chemical and biological tests were performed to characterize it. We have shown that the incorpration of fullerene into the hydrogel decreased the degradation and swelling rate of the hydrogel. A rheology test showed that the addition of fullerene increased the elastic modulus of the hydrogel. Chitosan hydrogel containing fullerene reduced the releasing rate of methotrexate (MTX) loaded in the hydrogel. A cell viability assay showed that fullerene in the hydrogel was not toxic to the cultured cells. The fullerene in the chitosan hydrogel significantly enhanced the anti-tumor cell effect. This study implies that the combined application of fullerene and chitosan hydrogel is a promising strategy for drug delivery in tumor therapy.