A novel cationic niosome formulation for gene delivery to the retina.
Niosomes represent a recent promising approach for gene delivery purposes. We elaborated on a novel niosome formulation based on the 2,3-di(tetradecyloxy)propan-1-amine cationic lipid, combined with squalene and polysorbate 80 to evaluate the transfection efficiency in rat retinas. Niosomes prepared by the solvent emulsification-evaporation technique were mixed with the pCMSEGFP plasmid to form lipoplexes which were characterized in terms of morphology, size, surface charge, and DNA condensation, protection and release. In vitro studies were conducted to evaluate transfection efficiency, viability and internalization mechanism in HEK-293 and ARPE-19 cells. The efficacy of the most promising formulation was evaluated in rat eyes by monitoring the expression of the EGFP after intravitreal and subretinal injections. Lipoplexes at 15/1 ratio were 200nm in size, 25mV in zeta potential and exhibited spherical morphology. At this ratio, niosomes condensed and protected the DNA from enzymatic digestion. Lipoplexes successfully transfected HEK-293 and specially ARPE-19 cells, without affecting the viability. Whereas lipoplexes entered mainly retinal cells by clathrin-mediated endocytosis, HEK-293 cells showed a higher caveolae-dependent entry. After ocular administration, the expression of EGFP was detected in different cells of the retina depending on the administration route. This novel niosome formulation represents a promising approach to deliver genetic material into the retina to treat inherited retinal diseases.
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain.
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Puras
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain.
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Mashal
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain.
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Zárate
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain.
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Agirre
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain.
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Ojeda
Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain; Institute of Advanced Chemistry of Catalonia, IQAC-CSIC, Barcelona, Spain.
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Grijalvo
Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain; Institute of Advanced Chemistry of Catalonia, IQAC-CSIC, Barcelona, Spain.
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Eritja
Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain; Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University, Elche, Spain.
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Diaz-Tahoces
Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain; Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University, Elche, Spain.
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Martínez Navarrete
Laboratory of Experimental Ophthalmology, Faculty of Medicine, University of Murcia, Regional Campus of International Excellence "Campus Mare Nostrum", Murcia, Spain.
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Avilés-Trigueros
Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain; Neuroprothesis and Neuroengineering Research Group, Miguel Hernández University, Elche, Spain.
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Fernández
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Centre of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza Spain. Electronic address: joseluis.pedraz@ehu.es.
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Pedraz
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D018014:Gene Transfer Techniques; D005995:Glyceryl Ethers / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008081:Liposomes; D008297:Male; D011437:Propylamines / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000378:metabolism*
D005995:Glyceryl Ethers; D008081:Liposomes; D011437:Propylamines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA
Cationic lipids; Niosomes; Non-ionic surfactant; Retina; Squalene; Transfection