摘要Cardiac arrest leads to global cerebral ischemia,causing significant neurological deficits with few effective treatments currently available.Neural stem cell(NSC)transplantation has shown therapeutic promise;however,challenges remain to achieve optimal and consistent delivery while maximizing cell functionality post-transplantation.Metabolic glycoengineering(MGE)has emerged as a novel technique to improve NSC viability and therapeutic efficacy by modifying their glycans.This report compares the effects of different delivery routes for this innovative approach of using MGE-modified NSCs(MGE-NSCs)to treat ischemic brain injury post-cardiac arrest.A total of 16 rats were subjected to 9-minute asphyxia cardiac arrest(4 weeks)and 21 rats subjected to 11-minute asphyxia cardiac arrest(3 days)to model severe and very severe brain injury,respectively.The MGE-NSCs were administered either intranasally or intracerebroventricularly(ICV)3 hours after resuscitation.Neurological deficits and neurobehavior tests were assessed periodically after resuscitation.Neuronal damage was examined with Fluro-Jade C staining and cresyl violet staining.The transplanted MGE-NSCs were tracked using immunofluorescence staining.In the early phase of recovery,the ICV administration resulted in better neurological deficit scores shortly after resuscitation compared with the intranasal route.Three days post-resuscitation,MGE-NSCs were primarily located in the cortex and hippocampus,with a higher percentage in the ICV-treated group.However,by 4 weeks,rats treated with intranasal MGE-NSCs exhibited superior locomotor function and reduced anxiety-/depression-related behaviors compared with those receiving ICV-NSC therapy.This was accompanied by greater survival and distribution of MGE-NSCs to the cerebellum and brainstem,along with reduced microglia recruitment.MGE-NSCs administered via both delivery methods enhanced vascular angiogenesis,neuron differentiation,and synaptic plasticity.Overall,intranasal delivery was as effective or potentially superior to ICV for long-term recovery,whereas ICV conferred short-term advantages.These results highlight the potential of MGE-NSC therapy to enhance post-cardiac arrest recovery,advocating for tailored treatment strategies based on specific recovery phase goals.