{"doi":"10.1016/j.xops.2025.100900","title":"Small Extracellular Vesicle–Mediated Peptide Delivery to the Mouse Corneal Endothelium In Vivo","abstract":"Purpose To investigate the utility of human corneal endothelial cell (hCEC)-derived small extracellular vesicles (sEV) as a delivery system to the mouse corneal endothelium (CE) in vivo . Methods hCEC-derived sEV were isolated from culture media using differential ultracentrifugation. Size, number, and morphology were characterized using nanoparticle tracking analysis and transmission electron microscopy (TEM). sEV markers were confirmed using an sEV detection antibody panel. PKH26-labeled sEV were injected into the anterior chamber of wild-type mice, and corneal endothelial uptake of sEV was assessed by colocalization of DAPI and PKH26 fluorescence on corneal whole mounts. Saponin permeabilization and sonication were used to load FITC-conjugated peptide into sEV. Delivery to mouse CE in vivo following intracameral injection was assessed by colocalization analysis of PKH26-labeled sEV with FITC-conjugated peptide. An average of 5 x 10 8 – 1 x 10 9 particles/plate with a peak size of 120 nm sEV were isolated. Results Morphology and size of sEV was confirmed by TEM, and the presence of sEV markers, FLOT1, ICAM, ALIX, CD81, CD63, ANXA5, and TSG101, was confirmed by immunoblotting. Confocal microscopy of whole-mounted mouse corneas, performed two days after intracameral injection of PKH26-labled sEV containing FITC-conjugated peptide, showed localization of sEV within the CE. Colocalization of PKH26 and FITC signals in the mouse CE confirmed delivery of the peptide cargo via hCEC-derived sEV in the mouse CE in vivo . Conclusion We demonstrate proof of principle of using hCEC-derived sEV for delivery of potential therapeutic agents to the mouse CE in vivo via intracameral injection.","journal":"Ophthalmology Science","year":2025,"id":545694,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9658,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":538091,"name":"Sun Young Lee","orcid":"0000-0003-3771-8799","position":1,"is_corresponding":false},{"id":857622,"name":"Dimitrios Pollalis","orcid":"0000-0001-9636-1967","position":2,"is_corresponding":false},{"id":331864,"name":"Martin Heur","orcid":"0000-0002-8965-6158","position":3,"is_corresponding":false},{"id":1144243,"name":"JeongGoo Lee","orcid":null,"position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-19T02:53:27.751050Z","pmid":"41049108","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}