not-yet-known not-yet-known not-yet-known unknown Traumatic Brain Injury (TBI) is a severe neurological disorder with an incomplete understanding of its underlying mechanisms, primarily due to the lack of effective strategy for in situ spatiotemporal analysis. Biomarkers associated with TBI, such as glial fibrillary acidic protein (GFAP), are typically detected in vitro rather than in situ, with a notable absence of spatiotemporal dynamics analysis. Herein, we developed a resveratrol-functionalized silver nanowires-doped MXene-based field-effect transistor biopatch (Res-Ag-MFETs) for in-situ spatiotemporal GFAP analysis, aiming to elucidate the TBI’s biomolecular mechanisms. We employed silver nanowires (AgNWs)-doped two-dimensional MXene as the FETs semiconductor and validated the favorable capability of MXene@AgNWs via morphological, elemental characterization, and DFT simulations. Res-Ag-MFETs demonstrated a favourable capability to suppress neuronal damage and inflammation, as evidenced by histological staining and bioactivity tests. Additionally, Res-Ag-MFETs demonstrated remarkable reproducibility (RSD=2.12%), stability, and sensitivity for GFAP quantification, achieving a detection limit as low as 0.47 pg/mL. Ultimately, Res-Ag-MFETs enabled efficient in-situ spatiotemporal analysis of GFAP in a Sprague Dawley (SD) rat with TBI, revealing a progressive diffusion of GFAP from the centre to the periphery over time. This advancement provides a novel platform for spatiotemporal dynamics analysis of biochemical markers in brain disorders, potentially laying the groundwork for further exploration of underlying pathogenic mechanisms.