Triboelectric nanogenerator (TENG) has shown great potential in real-time monitoring of individual physiological state. Conductive metal nanofibers can maintain good stability and mechanical strain as electrodes. Herein, a simple and safe strategy was provided to prepare electrospun gelatin membranes with in-situ coated silver layer for flexible TENG devices. Electrospun gelatin membranes were prepared by a one-pot method of dopamine-grafted gelatinmodification and physical modification by blending with polyvinyl alcohol (PVA). The silver coated electrospun membrane was constructed using the reductive immobilization of silver ions by dopamine. The silver content and hence the output signal of the TENG can be further controlled using bio-crosslinked gelatin membranes. The electrospun gelatin membranes of with immobilized silver layers with ease of preparation were not significantly cytotoxic to fibroblasts. The tensile strength of the membranes crosslinked reached 4MPa, which was more flexible than the membranes uncrosslinked. The membranes showed significant inhibition against Staphylococcus Aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) and the triboelectric output performance of the membrane can be controlled by adjusting the crosslinking method, which providing an effective strategy for implementing TENG in the monitoring of the wound healing process.