Preparation, characterization and evaluation of solid lipid nanoparticles and niosomes for ING4 gene delivery to MCF-7 cells

WOS: 000485773700017In this study, we aimed to develop lipid-based delivery systems for ING4 gene. For this purpose, we chose solid lipid nanoparticle (SLN) and niosome formulations for their advantages in several ways. SLNs were prepared via hot microemulsion method with slight modifications. Niosomes were prepared by using thin film hydration method. Both of those formulations contain cationic lipids for the purpose of gaining positive charge. Thus, complexation with ING4 gene carrier plasmid DNA is conducted via electrostatic interactions. DLS measurements showed us both of those formulations have appropriate zeta potential for complexation which is above 30 mV. Average particle size of SLNs is 9.76 +/- 0.13 nm. Vesicle size of niosomes is 483.2 +/- 20.19 nm. Both of those formulations can be considered nano-sized delivery systems. Reliability of those values were supported with TEM imaging. DNA protection ability of those formulations were observed in serum stability study. After characterization studies, cytotoxic effect of those delivery systems was evaluated on MCF-7 cell lines.Scientific Research Fund of Ege UniversityEge University [16ECZ025]This study was supported by the Scientific Research Fund of Ege University (project No: 16ECZ025)

In situ production of cationic lipid coated magnetic nanoparticles in multiple emulsions for gene delivery

WOS: 000379969900003Magnetic nanoparticles are effective delivery systems to target therapeutic genes by the attractive forces of magnetic fields. Curative effects depending on dose of nucleic acids or drugs increased, while cytotoxic effects minimized with these systems. In this study, a novel magnetic nanoparticle synthesis method was developed by combining advantages of microemulsion and multiple emulsion methods. Particle size, zeta potential, magnetization, complex formation with nucleic acids, DNase I protection ability, and cytotoxicity levels were examined. At last, magnetic nanoparticles were obtained with a promising synthesis method and it is determined that they are sufficiently small, non-toxic and have optimal surface properties for systemic delivery of nucleic acids.TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TUBITAK-SBAG-112S294]This study has been financially supported by TUBITAK under grant code TUBITAK-SBAG-112S294

Determining in vitro anticancer activity of mitomycin C loaded microemulsion

European Biotechnology Congress -- SEP 28-OCT 01, 2011 -- Istanbul, TURKEYWOS: 000295310800409European Biotechnol Themat Network Asso

PREPARATION AND CHARACTERISATION OF CATIONIC SOLID LIPID NANOPARTICLES AS A POTENTIAL DNA DELIVERY SYSTEM

WOS: 00020934590007

Development and in vitro evaluation of positive-charged solid lipid nanoparticles as nucleic acid delivery system in glioblastoma treatment

WOS: 000438786000019In this study, we aimed to develop a novel positive charged nucleic acid delivery system for the treatment of glioblastoma. For this purpose, Epidermal Growth Factor Receptor (EGFR) which plays a prominent role in glioblastoma was selected as a target. Cationic solid lipid nanoparticles (cSLN) were developed by microemulsion dilution method using cetyl palmitate as matrix lipid, Cremephor RH40 and Peceol as surfactants, and ethanol as cosurfactant. Characterization studies showed that obtained nanoparticles are positively charged and has an appropriate particle size for nucleic acid delivery (<20 nm). Gel retardation assay revealed that cSLNs have complexation ability with siRNA EGFR (cSLN:siRNAEGFR) and this complex is able to protect siRNA from serum- mediated degradation up to 6 h. The cytotoxicity evaluation of nanoparticles performed on U87 Human Glioblastoma Cell Line. Furthermore, in vitro delivery of siRNA-EGFR via cSLN inhibited EGFR expression significantly at 50nM siRNA dose compared to free siRNA-EGFR at the same dose on U87 cell line (p<0.05). Based on these findings, we propose that the developed cSLN system may has a potential as a siRNA delivery system for glioblastoma.Scientific Human Resources Development Program (OYP) of the Turkish Higher Education Council (YOK); TUBITAK-SBAGTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108S083]This work has been supported by Scientific Human Resources Development Program (OYP) of the Turkish Higher Education Council (YOK). We appreciate the project 108S083 (TUBITAK-SBAG) which the Malvern Zetasizer Instrument was purchased. Dr. Bilge Debelec Butuner from the Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Ege University is greatly appreciated for kindly providing the U87 cells

The effects of lyophilization and cryoprotectants on solid lipid nanoparticle-DNA systems

WOS: 000423282700020The development of optimized plasmid DNA delivery systems is necessary to deliver genetic material for effective gene therapy. To achieve this aim, a novel SLN: pDNA system was developed and the influence of the lyophilization procedure and the role of cryoprotectants in establishing the system's characteristics and stability were examined. SLNs were prepared by modified microemulsion dilution method. Afterward, a green fluorescent protein expression plasmid was loaded into the SLN system via electrostatic interactions. To examine the effects of cryoprotectants on the lyophilization process, sucrose, mannose, and trehalose were added at different ratios into the dilution medium; Tween 80 was also studied as an outer surfactant without cryoprotectants by incorporating it into the dilution medium. The system was characterized before and after the lyophilization procedure and tested with pDNA-loaded and unloaded SLNs. The compact form of the SLN: pDNA complexes were dissociated with cryoprotectants except %5 sucrose. Redispersing lyophilized SLNs and forming complexes with pDNA in an aseptic environment would be suitable at the application stage.Ege University, TurkeyEge University [09-ECZ-011]We are thankful to the Research Fund of Ege University, Turkey for financial support (Project No: 09-ECZ-011)

Development and evaluation of antisense shRNA-encoding plasmid loaded solid lipid nanoparticles against 5-alpha reductase activity

WOS: 000428980600030In this study, we aimed to develop and evaluate an effective system for nucleic acid delivery. For this purpose, shRNA-encoding plasmid against 5-alpha reductase (p5 alpha-Red) activity was selected as a model. In order to achieve the delivery of p5 alpha-Red and to obtain lower cytotoxicity, higher transfection and silencing efficiency, we developed solid lipid nanoparticles (SLNs) as a delivery vector by the melt-emulsification process. As a first step, microemulsions were prepared by using Compritol ATO 888 as internal oil phase; Tween 80 as a surfactant (S); ethanol as co-surfactant (CoS) and ultra-pure water as the continuous water phase. Then, obtained o/w microemulsion was dispersed in cold ultra-pure distilled water (0-4 degrees C) to form SLNs. The formulated nanoparticles were electrostatically bound to p5 alpha-Red to form SLN: p5 alpha-Red vectors. The SLN: p5 alpha-Red vectors have particle sizes of 62.65 nm, and zeta potential values of 12.9 mV. DNase I protection analysis showed that developed formulation is able to protect the p5 alpha-Red from degradation. The permeation study revealed that SLN: p5 alpha-Red vectors are able to pass the cellulose membrane and 80.2% of the SLN: p5 alpha-Red vectors were dialyzed after 6 h. According to cytotoxicity test results, no significant cytotoxicity was observed on DU-145 cells in the concentration range of 0.3-0.6 mu g/well. Furthermore, in vitro gene silencing experiment demonstrated that SLN: p5 alpha-Red vector effectively reduced 5 alpha-Red enzyme level 48 h after administration in DU-145 cell line. Considering the role of 5-alpha reductase in related diseases such as benign prostatic hyperplasia, androgenic alopecia and prostate cancer, the developed SLN: p5 alpha-Red vector system may have promises in future therapy.Ege University Research Fund [BAP] [14-ECZ-030]This work was supported by Ege University Research Fund [BAP, 14-ECZ-030, 2016]