Agradecimentos: The authors thank the financial support from Fundação de Amparo à pesquisa do Estado de São Paulo (São Paulo Research Foundation FAPESP – process number 2015/26701-0) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) finance code 001 (Coordination for the...

Agradecimentos: The authors thank the financial support from Fundação de Amparo à pesquisa do Estado de São Paulo (São Paulo Research Foundation FAPESP – process number 2015/26701-0) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) finance code 001 (Coordination for the Improvement of Higher Education Personnel). The authors kindly thank Professor Sávio Souza Venâncio Vianna and Arthur Lopes Valente for preparing Fig. 1 using ANSYS software. Authors also thank the Microfabrication Laboratory of Brazilian Center for Research in Energy and Materials (CNPEM) for the infrastructure to perform microfabrication as well as the Brazilian National Laboratory of Nanotechnology of CNPEM for the assistance in acquiring Cryo-TEM images of liposomes. The authors also want to thank the Brazilian Synchrotron Light Laboratory for beam times at SAXS1 beamline under research proposal (No. 20170804). Ismail Eş gratefully acknowledges the financial support of the São Paulo Research Foundation (FAPESP) (Grant # 2015/14468-0). Lucimara Gaziola de la Torre thanks Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq – National Council for Scientific and Technological Development) for the productivity grant (310735/2016-5).

Microfluidics is an emerging technology that allows efficient mixing of fluids on the nanoliter scale and has recently been employed for the production of lipid-based systems. In this study, we investigated the manufacturing of conventional (EPC/DOTAP/DOPE) and stealth...

Microfluidics is an emerging technology that allows efficient mixing of fluids on the nanoliter scale and has recently been employed for the production of lipid-based systems. In this study, we investigated the manufacturing of conventional (EPC/DOTAP/DOPE) and stealth (EPC/DOTAP/DOPE/DSPE-PEG(2000)) cationic liposomes (CLs) in a high-throughput chaotic advection-based microfluidic device (CA-MD) using water and ethanol. We assessed the effects of total flow rate (TFR), flow rate ratio (FRR), and flow configuration on CL formation. We compared the results with properties of CLs produced on a diffusion-based microfluidic device (D-MD). FRR (aqueous/solvent ratio) values were found to have a strong correlation with vesicle formation and values close to 1 led to the production of conventional and stealth CLs with diameter and polydispersity indices close to 200 nm and 0.2, respectively. Both liposomes are unilamellar, and stealth CL production could minimize micelle formation and operate at high flow velocities without altering characteristics of CLs. These findings were supported with dynamic light scattering (DLS), cryo-transmission electron microscopy (Cryo-TEM), and synchrotron small angle X-ray scattering (SAXS) techniques. The high solvent content in liposomes after microfluidic synthesis (FRR = 1) was reduced by employing centrifugal vacuum concentrators (CVC) at low pressure and 43 degrees C as an alternative distillation process for successful ethanol removal without changing the structural properties of these nanoaggregates. Finally, both CLs were able to transfect pDNA into PC-3 cancer cells. The CA-MD coupled to a CVC technique produced conventional and stealth CLs with high productivity, which makes it readily applicable for industrial production

CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ

310735/2016-5

COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES

FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP

2015/26701-0; 2015/14468-0

Fechado