Protamine/DNA/Niosome Ternary Nonviral Vectors for Gene Delivery to the Retina: The Role of Protamine.
The present study aimed to evaluate the incorporation of protamine into niosome/DNA vectors to analyze the potential application of this novel ternary formulation to deliver the pCMS-EGFP plasmid into the rat retina. Binary vectors based on niosome/DNA and ternary vectors based on protamine/DNA/niosomes were prepared and physicochemically characterized. In vitro experiments were performed in ARPE-19 cells. At 1:1:5 protamine/DNA/niosome mass ratio, the resulted ternary vectors had 150 nm size, positive charge, spherical morphology, and condensed, released, and protected the DNA against enzymatic digestion. The presence of protamine in the ternary vectors improved transfection efficiency, cell viability, and DNA condensation. After ocular administration, the EGFP expression was detected in different cell layers of the retina depending on the administration route without any sign of toxicity associated with the formulations. While subretinal administration transfected mainly photoreceptors and retinal pigment epithelial cells at the site of injection, intravitreal administration produced a more uniform distribution of the protein expression through the inner layers of the retina. The protein expression in the retina persisted for at least one month after both administrations. Our study highlights the flattering properties of protamine/DNA/niosome ternary vectors for efficient and safe gene delivery to the rat retina.
G
G
Puras
Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University , 03202 Alicante, Spain.
G
G
Martínez-Navarrete
M
M
Mashal
J
J
Zárate
M
M
Agirre
E
E
Ojeda
Institute for Advanced Chemistry of Catalonia, IQAC-CSIC, Spanish Council for Scientific Research , 08034 Barcelona, Spain.
S
S
Grijalvo
Institute for Advanced Chemistry of Catalonia, IQAC-CSIC, Spanish Council for Scientific Research , 08034 Barcelona, Spain.
R
R
Eritja
Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University , 03202 Alicante, Spain.
A
A
Diaz-Tahoces
Laboratory of Experimental Ophthalmology, Faculty of Medicine, University of Murcia , Regional Campus of International Excellence "Campus Mare Nostrum", E-30100 Murcia, Spain.
M
M
Avilés-Trigueros
Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University , 03202 Alicante, Spain.
E
E
Fernández
J L
JL
Pedraz
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D018014:Gene Transfer Techniques; D066298:In Vitro Techniques; D008081:Liposomes / Q000494:pharmacology / Q000627:therapeutic use*; D008297:Male; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D011479:Protamines / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D041623:Tomography, Optical Coherence; D014162:Transfection / Q000379:methods
D008081:Liposomes; D011479:Protamines; D004247:DNA
gene therapy; nanotechnology; niosomes; nonviral vectors; protamine; retina