Abstract
This chapter describes the synthesis of stealth and cationic liposomes and their complexation with plasmid DNA to generate lipoplexes for gene delivery applications. Two techniques are presented: a top-down approach which requires a second step of processing for downsizing the liposomes (i.e., ethanol injection method) and a microfluidic technique that explores the diffusion of ethanol in water to allow the proper lipid self-assembly. The synthesis of stealth liposomes is also a challenge since the use of poly(ethylene glycol) favors the formation of oblate micelles. In this protocol, the stealth cationic liposome synthesis by exploring the high ionic strength to overcome the formation of secondary structures like micelles is described. Finally, the electrostatic complexation between cationic liposomes and DNA is described, indicating important aspects that guarantee the formation of uniform lipoplexes.
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Kaleko M, Garcia JV, Miller AD (1991) Persistent gene expression after retroviral gene transfer into liver cells in vivo. Hum Gene Ther 32:27–32
de la Torre LG, Rosada RS, Trombone APF et al (2009) The synergy between structural stability and DNA-binding controls the antibody production in EPC/DOTAP/DOPE liposomes and DOTAP/DOPE lipoplexes. Colloids Surf B Biointerfaces 73:175–184
Verma IM, Weitzman MD (2005) Gene therapy: twenty-first century medicine. Annu Rev Biochem 74:711–738
Naldini L (2015) Gene therapy returns to centre stage. Nature 526:351–360
Zylberberg C, Gaskill K, Pasley S et al (2017) Engineering liposomal nanoparticles for targeted gene therapy. Gene Ther 24:441–452
Audouy S a L, de Leij LFMH, Hoekstra D et al (2002) In vivo characteristics of cationic liposomes as delivery vectors for gene therapy. Pharm Res 19:1599–1605
Sternberg B, Sorgi FL, Huang L (1994) New structures in complex formation between DNA and cationic liposomes visualized by freeze-fracture electron microscopy. FEBS Lett 356:361–366
Mönkkönen J, Urtti A (1998) Lipid fusion in oligonucleotide and gene delivery with cationic lipids. Adv Drug Deliv Rev 34:37–49
Parker AL, Newman C, Briggs S et al (2003) Nonviral gene delivery: techniques and implications for molecular medicine. Expert Rev Mol Med 5:1–15
Allen TM, Hansen C (1991) Pharmacokinetics of stealth versus conventional liposomes: effect of dose. BBA-Biomembranes 1068:133–141
Suk JS, Xu Q, Kim N et al (2016) PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev 99:28–51
Trevisan JE, Cavalcanti LP, Oliveira CL et al (2011) Technological aspects of scalable processes for the production of functional liposomes for gene therapy. In: Non-viral gene therapy. IntechOpen, Crotia. https://doi.org/10.5772/17869
Maulucci G, De Spirito M, Arcovito G et al (2005) Particle distribution in DMPC vesicles solutions undergoing different sonication times. Biophys J 88:3545–3550
De Paula Rigoletto T, Silva CL, Santana MHA et al (2012) Effects of extrusion, lipid concentration and purity on physico-chemical and biological properties of cationic liposomes for gene vaccine applications. J Microencapsul 29:759–769
Pupo E, Padrón A, Santana E et al (2005) Preparation of plasmid DNA-containing liposomes using a high-pressure homogenization-extrusion technique. J Control Release 104:379–396
Meure LA, Foster NR, Dehghani F (2008) Conventional and dense gas techniques for the production of liposomes: a review. AAPS PharmSciTech 9:798–809
Balbino TA, Aoki NT, Gasperini AAM et al (2013) Continuous flow production of cationic liposomes at high lipid concentration in microfluidic devices for gene delivery applications. Chem Eng J 226:423–433
Perli G, Pessoa ACSN, Balbino TA et al (2019) Ionic strength for tailoring the synthesis of monomodal stealth cationic liposomes in microfluidic devices. Colloids Surf B Biointerfaces 179:233–241
Eş I, Ok MT, Puentes-Martinez XE et al (2018) Evaluation of siRNA and cationic liposomes complexes as a model for in vitro siRNA delivery to cancer cells. Colloids Surf A Physicochem Eng Asp 555:280–289
Balbino TA, Gasperini AAM, Oliveira CLP et al (2012) Correlation of the physicochemical and structural properties of pDNA/cationic liposome complexes with their in vitro transfection. Langmuir 28:11535–11545
Vitor MT, Bergami-Santos PC, Piedade Cruz KS et al (2016) Dendritic cells stimulated by cationic liposomes. J Nanosci Nanotechnol 16:270–279
Gómez-Mascaraque LG, Casagrande Sipoli C, de La Torre LG et al (2017) Microencapsulation structures based on protein-coated liposomes obtained through electrospraying for the stabilization and improved bioaccessibility of curcumin. Food Chem 233:343–350
Wetzer B, Gerardo BYK, Frederic M et al (2001) Reducible cationic lipids for gene transfer. Biochem J 356:747–756
Balbino TA, Azzoni AR, de La Torre LG (2013) Microfluidic devices for continuous production of pDNA/cationic liposome complexes for gene delivery and vaccine therapy. Colloids Surf B Biointerfaces 111:203–210
Moreira NH, de Almeida AL d J, de Oliveira Piazzeta MH et al (2009) Fabrication of a multichannel PDMS/glass analytical microsystem with integrated electrodes for amperometric detection. Lab Chip 9:115–121
de Paula Rigoletto T, Zaniquelli MED, Santana MHA et al (2011) Surface miscibility of EPC/DOTAP/DOPE in binary and ternary mixed monolayers. Colloids Surf B Biointerfaces 83:260–269
Vitor MT, Bergami-Santos PC, Zômpero RHF et al (2017) Cationic liposomes produced via ethanol injection method for dendritic cell therapy. J Liposome Res 27:249–263
Toledo MAS, Janissen R, Favaro MTP et al (2012) Development of a recombinant fusion protein based on the dynein light chain LC8 for non-viral gene delivery. J Control Release 159:222–231
Eş I, Montebugnoli LJ, Filippi MFP et al (2020) High-throughput conventional and stealth cationic liposome synthesis using a chaotic advection-based microfluidic device combined with a centrifugal vacuum concentrator. Chem Eng J 382:122821
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de la Torre, L.G., Pessoa, A.C.S.N., de Carvalho, B.G., Taketa, T.B., Eş, I., Perli, G. (2021). Bulk and Microfluidic Synthesis of Stealth and Cationic Liposomes for Gene Delivery Applications. In: Sousa, Â. (eds) DNA Vaccines. Methods in Molecular Biology, vol 2197. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0872-2_14
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DOI: https://doi.org/10.1007/978-1-0716-0872-2_14
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