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Translating EVs\u2010based therapy from bench to bedside remains challenging due to inefficient delivery of EVs to the injured area and lack of techniques to visualize the entire targeting process. Here we developed a dopamine surface functionalization platform that facilitates easy and simultaneous conjugation of targeting peptide and multi\u2010mode imaging probes to the surface of EVs. Utilizing this platform we concurrently modified M2 microglia\u2010derived EVs (M2\u2010EVs) with neuronal targeting peptide rabies virus glycoprotein peptide 29 (RVG29) and multi\u2010modal imaging tracers, resulting in the targeted delivery of M2\u2010EVs to stroke mice brain and enabled the dynamic visualization of the targeting process from whole\u2010body to cellular levels. We determined that intra\u2010arterial injection achieved the highest efficiency of targeted delivery of engineered EVs to the stroke mice brain, improved therapeutic efficacy by reducing neuronal apoptosis. Mechanistically, EVs miRNA array revealed that a number of anti\u2010apoptosis related miRNAs were significantly up\u2010regulated, including miR\u2010221\u20103p and miR\u2010423\u20103p, both exerted anti\u2010apoptotic effects through p38\/ERK signalling pathways in stroke. Overall, this platform provides a facile and powerful tool for multifunctional engineering of EVs for multiscale therapeutic evaluation and enhancement of EV\u2010based therapy, with valuable prospects for clinical translation.<\/jats:p>","DOI":"10.1002\/jev2.70031","type":"journal-article","created":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T06:55:18Z","timestamp":1736405718000},"source":"Crossref","is-referenced-by-count":32,"title":["A simple polydopamine\u2010based platform for engineering extracellular vesicles with brain\u2010targeting peptide and imaging probes to improve stroke outcome"],"prefix":"10.1002","volume":"14","author":[{"given":"Xiaojing","family":"Shi","sequence":"first","affiliation":[{"name":"Shanghai Jiao Tong University Affiliated Sixth People's Hospital, School of Biomedical Engineering Shanghai Jiao Tong University  Shanghai China"},{"name":"McGovern Institute for Brain Research Massachusetts Institute of Technology  Cambridge Massachusetts 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