Biocompatible, biodegradable, and implantable energy generation materials are expected to play impor- tant roles in the use of transient self-powered systems in tissue engineering and other biomedical ap- plications. In particular, hyaluronic acid (HA) derived from mammals has attracted considerable atten- tion because of its biocompatibility, biodegradability, and tissue regenerating and wound healing abili- ties. Nevertheless, there has been little effort to utilize HA as the core material for energy generation. This study developed a simple but efficient method to prepare pure and cross-linked HA hydrogel films by solvent evaporation, and fabricate biocompatible, biodegradable, and high-performance triboelectric nanogenerators based on the HA hydrogel film (HA-TENGs). The peak-to-peak voltage and current out- puts of these HA-TENGs reached ~20 V and ~0.4 μA, respectively. Such power output was sufficient to light up several blue LEDs connected in series. The HA hydrogel film as a TENG material showed only minor degradation after long-term use. Furthermore, these HA hydrogel films exhibited no cytotoxicity at least within one day and even promoted the proliferation of MC3T3-E1 cells. Therefore, this method can be a powerful strategy to fabricate high-performance biocompatible, biodegradable energy devices for a range of practical tissue engineering and other biomedical applications in the future.