The rapid development of wearable and self-powered technologies has led to high demand for sustainable, low-cost energy-harvesting solutions. In this study, a triboelectric nanogenerator (TENG) was fabricated using discarded disposable plates as the positive triboelectric layer and waste nitrile gloves as the negative layer, demonstrating an effective method for converting waste into functional devices. The TENG worked in the single-electrode mode, producing a maximum output voltage of 170 V and a current of 240 nA. A peak power output of 32 µW with an external load resistance of 500 MΩ was achieved using the fabricated TENG. Because of its consistent electrical performance, the TENG successfully powered small electronic devices such as a digital watch and a calculator. In addition to energy harvesting, the generated electrical output was used to stimulate biological activity in vitro, thereby increasing the migration of NIH/3T3 fibroblasts. Furthermore, the TENG could convert bodily movements into electrical energy. The combination of biomechanical sensing and electrically aided cell migration demonstrates the TENG’s potential as a long-term platform for self-powered biomedical devices.