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Triboelectric nanogenerators (TENGs) have become reliable green energy harvesters by converting biomechanical motions into electricity. However, the inevitable charge leakage and poor electric field (EF) of conventional TENG result in inferior tribo-charge density on the active layer. In this paper, TiO 2 -MXene incorporated polystyrene (PS) nanofiber membrane (PTMx NFM) charge trapping interlayer is introduced into single electrode mode TENG (S-TENG) to prevent electron loss at the electrode interface. Surprisingly, this charge-trapping mechanism augments the surface charge density and electric output performance of TENGs. Polyvinylidene difluoride (PVDF) mixed polyurethane (PU) NFM is used as tribo-active layer, which improves the crystallinity and mechanical property of PVDF to prevent delamination during long cycle tests. Herein, the effect of this double-layer capacitive model is explained experimentally and theoretically. With optimization of the PTMx interlayer thickness, S-TENG exhibits a maximum open-circuit voltage of (280 V), short-circuit current of (20 µA) transfer charge of (120 nC), and power density of (25.2 µW cm -2 ). Then, this energy is utilized to charge electrical appliances. In addition, the influence of AC/DC EF simulation in wound healing management (vitro L929 cell migration, vivo tissue regeneration) is also investigated by changing the polarity of trans-epithelial potential (TEP) distribution in the wounded area., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)