Objective: The aim of this study was to evaluate the neurorestorative effects of ultrasound targeted microbubble destruction (UTMD) in conjunction with the co-transplantation of astrocytes (AS) and neural stem cells (NSCs) on a murine model of traumatic brain injury (TBI). Methods: Fifty mice were randomly allocated into five groups: a control group, a TBI group, a UTMD group, a UTMD+NSCs group (UN), and a UTMD+NSCs+AS group (UNA). Treatment commenced one day post-TBI induction, with subsequent administrations every three days for a total of three treatments. Neurological deficits were assessed using the modified neurological severity score (mNSS) at 1, 4, 8, and 12 days post-treatment. On day 12, brain tissue edema was quantified using magnetic resonance imaging (MRI), and the levels of neuroprotective factors, including brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF), and nerve growth factor (NGF), as well as damage-associated factors such as heat shock protein 70 (HSP70), central nervous system-specific protein (S100B), and caspase-3, were measured using enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). Results: The UNA group showed significantly lower mNSS scores compared to the TBI and UTMD groups on day 8 of treatment (P<0.05). By day 12, the UNA group exhibited a significant reduction in mNSS scores compared to all other groups (P<0.05). Additionally, the UNA group demonstrated a significantly reduced brain tissue edema volume and differential expression of BDNF and caspase-3, with statistically significant differences observed compared to other groups (P<0.05). Conclusion: UTMD-mediated co-transplantation of astrocytes and neural stem cells significantly improved the biochemical microenvironment in TBI mice, upregulated the expression of neuroprotective factors, downregulated the expression of damage-associated factors, and enhanced the recovery of neurological function.