陆军军医大学学报2026，Vol.48Issue(4)：407-419,13.DOI:10.16016/j.2097-0927.202511021
通过系统性阳离子材料筛选优化牛奶外泌体载体以实现mRNA的高效肺部靶向递送
Optimizing milk-derived exosome carriers through systematic cationic material screening for efficient pulmonary-targeted mRNA delivery
摘要
Abstract
Objective To construct an engineered milk-derived exosome(mExos)vector for efficient messenger RNA(mRNA)delivery to the respiratory tract by systematically screening cationic modification materials.Methods mExos were isolated via ultracentrifugation,followed by electroporation-mediated loading of firefly luciferase(Fluc)or enhanced green fluorescent protein(EGFP)mRNA.Chitosan(CS),polyethyleneimine(PEI),poly(beta-amino ester)(PBAE),cationic lipid(DOTAP),and protamine were used to modify mRNA-loaded mExos.Dynamic light scattering,nanoparticle tracking analysis,flow cytometry and luciferase reporter assay were employed to evaluate particle size,zeta potential,colloidal stability,cellular uptake,transfection efficiency,and cytotoxicity.Fifty female BALB/c mice(6 to 8 weeks,about 20 g)received intranasal administration of dye-labeled or Fluc-mRNA-loaded formulations.In vivo distribution and pulmonary transfection kinetics were monitored by IVIS imaging,while histopathology and serum biochemistry of major organs were analyzed at 24 h post-administration.Results Cationic-modified mExos were successfully constructed.CS-modified mExos(CS-mExos)demonstrated surface charge reversal while maintaining optimal size and stability(PDI unchanged within 48 h).In vitro,CS-mExos enhanced cellular uptake in 16HBE and A549 cells by 2.56-fold(P＜0.001)and 4.56-fold(P＜0.001),respectively,and increased Fluc-mRNA transfection efficiency by 28.9-fold(P＜0.001)and 26.5-fold(P＜0.001)versus unmodified mExos,with＞95%cell viability.In vivo imaging revealed CS-mExos specifically accumulated in the lungs with sustained retention＞72 h,mediating 9.5-fold higher peak Fluc-mRNA expression at 6 h post-administration.Histopathology and serum biochemistry confirmed no significant organ damage or abnormal hepatic/renal indices.Conclusion Systematic screening identified chitosan as the optimal cationic material for engineering milk exosomes.The developed CS-mExos vector enables efficient targeted mRNA delivery to the respiratory tract with enhanced pulmonary transfection and favorable biosafety.关键词
牛奶外泌体/壳聚糖/信使RNA/肺部给药/基因治疗Key words
milk-derived exosomes/chitosan/messenger RNA/pulmonary administration/genetic therapy分类
医药卫生引用本文复制引用
罗明星,廖睿,关山,张卫军,罗萍,张怡,程平,李飏,邹全明..通过系统性阳离子材料筛选优化牛奶外泌体载体以实现mRNA的高效肺部靶向递送[J].陆军军医大学学报,2026,48(4):407-419,13.基金项目
国家自然科学基金面上项目(82173764,32370993) （82173764,32370993）
重庆市自然科学基金面上项目(CSTB2025NSCQ-GPX0624) Supported by the General Program of National Natural Science Foundation of China(82173764,32370993)and the General Program of Natural Science Foundation of Chongqing(CSTB2025NSCQ-GPX0624). （CSTB2025NSCQ-GPX0624）
