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MICROFLUIDIC SYSTEMS FOR INCORPORATION OF SMALL INTERFERING RNA (siRNA) IN CATIONIC LIPOSOMES AND FOR IN VITRO ANIMAL CELL TRANSFECTION TARGETING GENE THERAPY

Grant number: 15/14468-0
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): October 01, 2015
Effective date (End): August 31, 2020
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Lucimara Gaziola de la Torre
Grantee:Ismail Es
Host Institution: Faculdade de Engenharia Química (FEQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated scholarship(s):18/23895-7 - Microfluidic design for in vitro transfection of multicellular spheroids of HeLa cells using lipid-based gene delivery system, BE.EP.DR

Abstract

This research project aims the technological development of microfluidic systems to incorporate small interfering RNA (siRNA-silencer) inside of nano-scaled cationic liposomes (CLs), in order to get non-viral vectors for applications in gene therapy. Gene therapy is a promising technique that introduces a correct copy of the defective gene for the treatment of genetic diseases. The production of CLs-siRNA complexes by using microfluidics technology, which processes fluids in small amounts, enables the continuous operation as a single-phase system. The project will be divided into three stages: In the first stage, microfluidic device will be designed and the electrostatic complexation between conventional CLs and siRNA in microchannels will be studied and compared to bulk process, which is a conventional technique to produce CLs/siRNA complexes. In the second stage, the liposome composition will be modified with the inclusion of lipid derivatives that contain polyethylene glycol-distearoyl phosphatidylethanolamine with folate (DSPE-PEG-Folate) in order to generate Stealth liposomes with specific targeting to cells with folate receptors on their surface. The efficiency of the new liposome composition will be compared with the conventional ones. At the end of the first two stages, the physicochemical and biochemical properties of the complexes will be evaluated. In the final stage, the most appropriate lipid structures containing siRNA that will have been produced in previous stages will be transfected as a dynamic transfection analysis within microfluidic systems into HeLa Reporter Cells (HeLa) that stably express Green Fluorescent Protein (GFP). The expression level of GFP will be visualized and evaluated by using fluorescence microscope and afterwards, the transfection efficiency will be compared with the conventional method carried out in wells.

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Scientific publications (9)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MOAZZEN, MOJTABA; KHANEGHAH, AMIN MOUSAVI; SHARIATIFAR, NABI; AHMADLOO, MAHSA; ES, ISMAIL; BAGHANI, ABBAS NOROUZIAN; YOUSEFINEJAD, SAEED; ALIMOHAMMADI, MAHMOOD; AZARI, ALI; DOBARADARAN, SINA; et al. Multi-walled carbon nanotubes modified with iron oxide and silver nanoparticles (MWCNT-Fe3O4/Ag) as a novel adsorbent for determining PAEs in carbonated soft drinks using magnetic SPE-GC/MS method. ARABIAN JOURNAL OF CHEMISTRY, v. 12, n. 4, p. 476-488, . (15/14468-0)
ES, ISMAIL; MALFATTI-GASPERINI, ANTONIO A.; DE LA TORRE, LUCIMARA GAZIOLA. The diffusion-driven microfluidic process to manufacture lipid-based nanotherapeutics with stealth properties for siRNA delivery. COLLOIDS AND SURFACES B-BIOINTERFACES, v. 215, p. 11-pg., . (15/14468-0, 18/19537-8)
HASHEMI, SEYED MOHAMMAD BAGHER; KHANEGHAH, AMIN MOUSAVI; GHAHFARROKHI, MARYAM GHADERI; ES, ISMAIL. Basil-seed gum containing Origanum vulgare subsp viride essential oil as edible coating for fresh cut apricots. Postharvest Biology and Technology, v. 125, p. 26-34, . (15/14468-0)
ES, ISMAIL; KHANEGHAH, AMIN MOUSAVI; HASHEMI, SEYED MOHAMMAD BAGHER; KOUBAA, MOHAMED. Current advances in biological production of propionic acid. Biotechnology Letters, v. 39, n. 5, p. 635-645, . (15/14468-0)
ES, ISMAIL; OK, MERYEM TYRRASCH; PUENTES-MARTINEZ, XIMENA E.; SZYMANSKI DE TOLEDO, MARCELO AUGUSTO; DE PINHO FAVARO, MARIANNA TEIXEIRA; CAVALCANTI, LEIDE PASSOS; CASSAGO, ALEXANDRE; PORTUGAL, RODRIGO VILLARES; AZZONI, ADRIANO RODRIGUES; DE LA TORRE, LUCIMARA GAZIOLA. Evaluation of siRNA and cationic liposomes complexes as a model for in vitro siRNA delivery to cancer cells. COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS, v. 555, p. 280-289, . (15/14468-0)
KHANEGHAH, AMIN MOUSAVI; HASHEMI, SEYED MOHAMMAD BAGHER; ES, ISMAIL; FRACASSETTI, DANIELA; LIMBO, SARA. Efficacy of Antimicrobial Agents for Food Contact Applications: Biological Activity, Incorporation into Packaging, and Assessment Methods: A Review. JOURNAL OF FOOD PROTECTION, v. 81, n. 7, p. 1142-1156, . (15/14468-0)
ES, ISMAIL; MONTEBUGNOLI, LEONARDO JOSE; FILIPPI, MARIA FERNANDA P.; MALFATTI-GASPERINI, ANTONIO A.; RADAIC, ALLAN; DE JESUS, MARCELO BISPO; DE LA TORRE, LUCIMARA GAZIOLA. High-throughput conventional and stealth cationic liposome synthesis using a chaotic advection-based microfluidic device combined with a centrifugal vacuum concentrator. CHEMICAL ENGINEERING JOURNAL, v. 382, . (15/14468-0, 15/26701-0)
HASHEMI, SEYEDMOHAMMAD BAGHER; KHANEGHAH, AMIN MOUSAVI; KONTOMINAS, MICHAEL G.; ES, ISMAIL; SANT'ANA, ANDERSON S.; MARTINEZ, RAFAEL R.; DRIDER, DJAMEL. Fermentation of sarshir (kaymak) by lactic acid bacteria: antibacterial activity, antioxidant properties, lipid and protein oxidation and fatty acid profile. Journal of the Science of Food and Agriculture, v. 97, n. 13, p. 4595-4603, . (15/14468-0)
ES, ISMAIL; KHANEGHAH, AMIN MOUSAVI; BARBA, FRANCISCO J.; SARAIVA, JORGE A.; SANT'ANA, ANDERSON S.; BAGHER HASHEMI, SEYED MOHAMMAD. Recent advancements in lactic acid production - a review. Food Research International, v. 107, p. 763-770, . (15/14468-0)

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Filed patent(s) as a result of this research project

PROCESSO MICROFLUÍDICO DE OBTENÇÃO DE LIPOSSOMAS STEALTH CATIÔNICOS BR1320180744827 - Universidade Estadual de Campinas (UNICAMP) . LUCIMARA GAZIOLA DE LA TORRE / ISMAIL ES / LEONARDO JOSE MONTEBUGNOLI / ALLAN RADAIC / MARCELO BISPO DE JESUS - November 2018, 27