Cationic core-shell liponanoparticles for ocular gene delivery.
To achieve enhanced gene transfection efficiency with ocular eye-drop therapy, a cationic core-shell liponanoparticle (DLCS-NP) was designed by enveloping the plasmid-laden chitosan nanoparticle (CS-NP) into a cationic lipid shell. The cellular uptake of DLCS-NP was up to 1.25-fold and 5-fold higher than that of CS-NP and lipid-coated chitosan nanoparticles (LCS-NP), respectively. Further endocytosis inhibition investigation discovered that facilitated by the cationic outer lipid layer, several other distinct pathways (besides clathrin-mediated endocytosis) were involved in the endocytosis of DLCS-NP. Endolysosome trafficking experiment verified that cationic lipid coating could facilitate the endolysosome escape of DLCS-NP. Consequently, using enhanced green fluorescence protein (EGFP) as a reporter gene, DLCS-NP-treated human conjunctival epithelial cells exhibited 3.1- and 3.5-fold more intense EGFP expression than that of LCS-NP and CS-NP, respectively. Finally, in vivo transfection experiments on rabbits revealed that EGFP expression exhibited 2.52-fold increase in DLCS-NP group than that of CS-NP group. In summary, this type of cationic core-shell liponanoparticle, possessing multiple functions including better DNA protecting effect, superior cellular uptake efficiency, utilization of multiple endocytic pathways, and endolysosome escaping ability, may represent a promising strategy for ocular gene delivery.
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Min
M
Jiang
Li
L
Gan
Chunliu
C
Zhu
Yang
Y
Dong
Jianping
J
Liu
Yong
Y
Gan
D000818:Animals; D002412:Cations; D016923:Cell Death; D002470:Cell Survival; D024202:Electrophoretic Mobility Shift Assay; D004705:Endocytosis; D005123:Eye / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008247:Lysosomes / Q000378:metabolism; D008297:Male; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011817:Rabbits; D055672:Static Electricity; D014162:Transfection
D002412:Cations; D008055:Lipids; D049452:Green Fluorescent Proteins; D016650:Fluorescein-5-isothiocyanate