Integrated microfluidic devices for the synthesis of nanoscale liposomes and lipoplexes.
In this work, pDNA/cationic liposome (CL) lipoplexes for gene delivery were prepared in one-step using multiple hydrodynamic flow-focusing regions. The microfluidic platform was designed with two distinct regions for the synthesis of liposomes and the subsequent assembly with pDNA, forming lipoplexes. The obtained lipoplexes exhibited appropriate physicochemical characteristics for gene therapy applications under varying conditions of flow rate-ratio (FRR), total volumetric flow rate (QT) and pDNA content (molar charge ratio, R±). The CLs were able to condense and retain the pDNA in the vesicular structures with sizes ranging from 140nm to 250nm. In vitro transfection assays showed that the lipoplexes prepared in one step by the two-stage configuration achieved similar efficiencies as lipoplexes prepared by conventional bulk processes, in which each step comprises a series of manual operations. The integrated microfluidic platform generates lipoplexes with liposome formation combined in-line with lipoplex assembly, significantly reducing the number of steps usually required to form gene carrier systems.
Department of Materials Engineering and Bioprocess, School of Chemical Engineering, University of Campinas, UNICAMP, São Paulo, SP, 13083-970, Brazil.
Tiago A
TA
Balbino
Department of Materials Engineering and Bioprocess, School of Chemical Engineering, University of Campinas, UNICAMP, São Paulo, SP, 13083-970, Brazil.
Juliana M
JM
Serafin
Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, UNICAMP, São Paulo, SP, 13083-862, Brazil.
Allan
A
Radaic
Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, UNICAMP, São Paulo, SP, 13083-862, Brazil.
Marcelo B
MB
de Jesus
Department of Materials Engineering and Bioprocess, School of Chemical Engineering, University of Campinas, UNICAMP, São Paulo, SP, 13083-970, Brazil. Electronic address: latorre@feq.unicamp.br.
Lucimara G
LG
de la Torre
D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D008081:Liposomes / Q000737:chemistry*; D044085:Microfluidics / Q000379:methods*; D010957:Plasmids / Q000737:chemistry
D008081:Liposomes; D004247:DNA
DNA; Gene delivery; Lipoplexes; Liposomes; Microfluidics; Nanocarriers