Efficient intracellular siRNA delivery strategy through rapid and simple two steps mixing involving noncovalent post-PEGylation.
Two different and well-defined methacrylate-based (co)polymers were employed as a polymeric siRNA delivery system. siRNA, poly(2-(dimethylamino) ethyl methacrylate) homopolymers (PDMAEMA) and poly(2-(dimethylamino) ethyl methacrylate)-b-poly (ethyleneglycol) alpha-methoxy, omega-methacrylate (PDMAEMA-b-PMAPEG) palm-tree-like copolymer ternary complexes were prepared using a rapid and simple two-step mixing protocol involving noncovalent post-PEGylation, and physicochemical properties including hydrodynamic diameter, zeta-potential and siRNA condensation efficiency were characterized. Transfection efficiency, intracellular uptake, and cytotoxicity of ternary complexes were also evaluated. Ternary complexes provide efficient condensation and compaction of siRNA within the cationic core of complexes. Noncovalent post-PEGylation provides the ternary complexes with enzymatic and serum stability without harming complex formation and condensation of siRNA. Thereby, under an optimal N/P ratio, ternary complexes exhibited brilliant gene silencing efficiency with low cytotoxicity in media containing 10% serum. Confocal microscopy clearly showed efficient and even intracellular uptake of complexes by cells via endocytosis. This study highlights the excellent properties of noncovalent post-PEGylated ternary complexes produced by rapid and simple mixing. Accordingly, these findings suggest that the formation of ternary complexes could be utilized as a safe and effective polymeric siRNA delivery strategy.
Kong
Won-Ho
WH
Biomedical Engineering Interdisciplinary Course, Sungkyunkwan University School of Medicine, 50 Irwon-Dong, Gangnam-Gu, Seoul 135-710, Republic of Korea.
Sung
Dong-Kyung
DK
Shim
Yong-Ho
YH
Bae
Ki Hyun
KH
Dubois
Philippe
P
Park
Tae Gwan
TG
Kim
Jin-Hoon
JH
Seo
Soo-Won
SW
D000180:Acrylic Resins / Q000737:chemistry*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry*; D016503:Drug Delivery Systems / Q000379:methods; D004355:Drug Stability; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D008297:Male; D008689:Methacrylates / Q000737:chemistry*; D018613:Microscopy, Confocal; D008958:Models, Molecular; D009757:Nylons / Q000737:chemistry*; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
D000180:Acrylic Resins; D004337:Drug Carriers; D008689:Methacrylates; D009757:Nylons; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; C524500:poly(2-(dimethylamino) ethyl methacrylate)-b-poly(polyethylene glycol methacrylate); C407037:poly(2-(dimethylamino)ethyl methacrylate); D011092:Polyethylene Glycols