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Sample records for nanoparticles transfection study

  1. Thiolated chitosan nanoparticles: transfection study in the Caco-2 differentiated cell culture

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    Martien, Ronny [Department of Pharmaceutical Technology, Institute of Pharmacy, Leopold-Franzens-University of Innsbruck, Innrain 52, Josef Moeller Haus, A-6020 Innsbruck (Austria); Loretz, Brigitta [Department of Pharmaceutical Technology, Institute of Pharmacy, Leopold-Franzens-University of Innsbruck, Innrain 52, Josef Moeller Haus, A-6020 Innsbruck (Austria); Sandbichler, Adolf Michael [Institute of Zoology, Leopold-Franzens-University of Innsbruck, Technikerstrasse 25, A-6020 Innsbruck (Austria); Schnuerch, Andreas Bernkop [Department of Pharmaceutical Technology, Institute of Pharmacy, Leopold-Franzens-University of Innsbruck, Innrain 52, Josef Moeller Haus, A-6020 Innsbruck (Austria)

    2008-01-30

    The aim of this study was to monitor the expression of secreted protein in differentiated Caco-2 cells after transfection with nanoparticles, in order to improve gene delivery. Based on unmodified chitosan and thiolated chitosan conjugates, nanoparticles with the gene reporter pSEAP (recombinant Secreted Alkaline Phosphatase) were generated at pH 4.0. Transfection studies of thiolated chitosan in Caco-2 cells during the exponential growth phase and differentiation growth phase of the cells led to a 5.0-fold and 2.0-fold increase in protein expression when compared to unmodified chitosan nanoparticles. The mean particle size for both unmodified chitosan and cross-linked thiolated chitosan nanoparticles is 212.2 {+-} 86 and 113.6 {+-} 40 nm, respectively. The zeta potential of nanoparticles was determined to be 7.9 {+-} 0.38 mV for unmodified chitosan nanoparticles and 4.3 {+-} 0.74 mV for cross-linked thiolated chitosan nanoparticles. Red blood cell lysis evaluation was used to evaluate the membrane damaging properties of unmodified and thiolated chitosan nanoparticles and led to 4.61 {+-} 0.36% and 2.29 {+-} 0.25% lysis, respectively. Additionally, cross-linked thiolated chitosan nanoparticles were found to exhibit higher stability toward degradation in gastric juices. Furthermore the reversible effect of thiolated chitosan on barrier properties was monitored by measuring the transepithelial electrical resistance (TEER) and is supported by immunohistochemical staining for the tight junction protein claudin. According to these results cross-linked thiolated chitosan nanoparticles have the potential to be used as a non-viral vector system for gene therapy.

  2. Thiolated chitosan nanoparticles: transfection study in the Caco-2 differentiated cell culture

    International Nuclear Information System (INIS)

    Martien, Ronny; Loretz, Brigitta; Sandbichler, Adolf Michael; Schnuerch, Andreas Bernkop

    2008-01-01

    The aim of this study was to monitor the expression of secreted protein in differentiated Caco-2 cells after transfection with nanoparticles, in order to improve gene delivery. Based on unmodified chitosan and thiolated chitosan conjugates, nanoparticles with the gene reporter pSEAP (recombinant Secreted Alkaline Phosphatase) were generated at pH 4.0. Transfection studies of thiolated chitosan in Caco-2 cells during the exponential growth phase and differentiation growth phase of the cells led to a 5.0-fold and 2.0-fold increase in protein expression when compared to unmodified chitosan nanoparticles. The mean particle size for both unmodified chitosan and cross-linked thiolated chitosan nanoparticles is 212.2 ± 86 and 113.6 ± 40 nm, respectively. The zeta potential of nanoparticles was determined to be 7.9 ± 0.38 mV for unmodified chitosan nanoparticles and 4.3 ± 0.74 mV for cross-linked thiolated chitosan nanoparticles. Red blood cell lysis evaluation was used to evaluate the membrane damaging properties of unmodified and thiolated chitosan nanoparticles and led to 4.61 ± 0.36% and 2.29 ± 0.25% lysis, respectively. Additionally, cross-linked thiolated chitosan nanoparticles were found to exhibit higher stability toward degradation in gastric juices. Furthermore the reversible effect of thiolated chitosan on barrier properties was monitored by measuring the transepithelial electrical resistance (TEER) and is supported by immunohistochemical staining for the tight junction protein claudin. According to these results cross-linked thiolated chitosan nanoparticles have the potential to be used as a non-viral vector system for gene therapy

  3. Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection

    Science.gov (United States)

    Kalies, Stefan; Heinemann, Dag; Schomaker, Markus; Gentemann, Lara; Meyer, Heiko; Ripken, Tammo

    2014-01-01

    Abstract. In comparison to standard transfection methods, gold nanoparticle-mediated laser transfection has proven to be a versatile alternative. This is based on its minor influence on cell viability and its high efficiency, especially for the delivery of small molecules like small interfering RNA. However, in order to transfer it to routine usage, a safety aspect is of major concern: The avoidance of nanoparticle uptake by the cells is desired. The immobilization of the gold nanoparticles on cell culture surfaces can address this issue. In this study, we achieved this by silanization of the appropriate surfaces and the binding of gold nanoparticles to them. Comparable perforation efficiencies to the previous approaches of gold nanoparticle-mediated laser transfection with free gold nanoparticles are demonstrated. The uptake of the immobilized particles by the cells is unlikely. Consequently, these investigations offer the possibility of bringing gold nanoparticle-mediated laser transfection closer to routine usage. PMID:25069006

  4. Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection.

    Science.gov (United States)

    Kalies, Stefan; Heinemann, Dag; Schomaker, Markus; Gentemann, Lara; Meyer, Heiko; Ripken, Tammo

    2014-01-01

    In comparison to standard transfection methods, gold nanoparticle-mediated laser transfection has proven to be a versatile alternative. This is based on its minor influence on cell viability and its high efficiency, especially for the delivery of small molecules like small interfering RNA. However, in order to transfer it to routine usage, a safety aspect is of major concern: The avoidance of nanoparticle uptake by the cells is desired. The immobilization of the gold nanoparticles on cell culture surfaces can address this issue. In this study, we achieved this by silanization of the appropriate surfaces and the binding of gold nanoparticles to them. Comparable perforation efficiencies to the previous approaches of gold nanoparticle-mediated laser transfection with free gold nanoparticles are demonstrated. The uptake of the immobilized particles by the cells is unlikely. Consequently, these investigations offer the possibility of bringing gold nanoparticle-mediated laser transfection closer to routine usage.

  5. Comparison of nanoparticle-mediated transfection methods for DNA expression plasmids: efficiency and cytotoxicity

    Science.gov (United States)

    2011-01-01

    Background Reproducibly high transfection rates with low methodology-induced cytotoxic side effects are essential to attain the required effect on targeted cells when exogenous DNA is transfected. Different approaches and modifications such as the use of nanoparticles (NPs) are being evaluated to increase transfection efficiencies. Several studies have focused on the attained transfection efficiency after NP-mediated approaches. However, data comparing toxicity of these novel approaches with conventional methods is still rare. Transfection efficiency and methodology-induced cytotoxicity were analysed after transfection with different NP-mediated and conventional approaches. Two eukaryotic DNA-expression-plasmids were used to transfect the mammalian cell line MTH53A applying six different transfection protocols: conventional transfection reagent (FuGENE HD, FHD), FHD in combination with two different sizes of stabilizer-free laser-generated AuNPs (PLAL-AuNPs_S1,_S2), FHD and commercially available AuNPs (Plano-AuNP), and two magnetic transfection protocols. 24 h post transfection efficiency of each protocol was analysed using fluorescence microscopy and GFP-based flow cytometry. Toxicity was assessed measuring cell proliferation and percentage of propidium iodide (PI%) positive cells. Expression of the respective recombinant proteins was evaluated by immunofluorescence. Results The addition of AuNPs to the transfection protocols significantly increased transfection efficiency in the pIRES-hrGFPII-eIL-12 transfections (FHD: 16%; AuNPs mean: 28%), whereas the magnet-assisted protocols did not increase efficiency. Ligand-free PLAL-AuNPs had no significant cytotoxic effect, while the ligand-stabilized Plano-AuNPs induced a significant increase in the PI% and lower cell proliferation. For pIRES-hrGFPII-rHMGB1 transfections significantly higher transfection efficiency was observed with PLAL-AuNPs (FHD: 31%; PLAL-AuNPs_S1: 46%; PLAL-AuNPs_S2: 50%), while the magnet

  6. DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons.

    Science.gov (United States)

    Vernon, Matthew M; Dean, David A; Dobson, Jon

    2015-08-17

    Efficient non-viral plasmid DNA transfection of most stem cells, progenitor cells and primary cell lines currently presents an obstacle for many applications within gene therapy research. From a standpoint of efficiency and cell viability, magnetic nanoparticle-based DNA transfection is a promising gene vectoring technique because it has demonstrated rapid and improved transfection outcomes when compared to alternative non-viral methods. Recently, our research group introduced oscillating magnet arrays that resulted in further improvements to this novel plasmid DNA (pDNA) vectoring technology. Continued improvements to nanomagnetic transfection techniques have focused primarily on magnetic nanoparticle (MNP) functionalization and transfection parameter optimization: cell confluence, growth media, serum starvation, magnet oscillation parameters, etc. Noting that none of these parameters can assist in the nuclear translocation of delivered pDNA following MNP-pDNA complex dissociation in the cell's cytoplasm, inclusion of a cassette feature for pDNA nuclear translocation is theoretically justified. In this study incorporation of a DNA targeting sequence (DTS) feature in the transfecting plasmid improved transfection efficiency in model neurons, presumably from increased nuclear translocation. This observation became most apparent when comparing the response of the dividing SH-SY5Y precursor cell to the non-dividing and differentiated SH-SY5Y neuroblastoma cells.

  7. Gold nanoparticle mediated laser transfection for efficient siRNA mediated gene knock down.

    Directory of Open Access Journals (Sweden)

    Dag Heinemann

    Full Text Available Laser based transfection methods have proven to be an efficient and gentle alternative to established molecule delivery methods like lipofection or electroporation. Among the laser based methods, gold nanoparticle mediated laser transfection bears the major advantage of high throughput and easy usability. This approach uses plasmon resonances on gold nanoparticles unspecifically attached to the cell membrane to evoke transient and spatially defined cell membrane permeabilization. In this study, we explore the parameter regime for gold nanoparticle mediated laser transfection for the delivery of molecules into cell lines and prove its suitability for siRNA mediated gene knock down. The developed setup allows easy usage and safe laser operation in a normal lab environment. We applied a 532 nm Nd:YAG microchip laser emitting 850 ps pulses at a repetition rate of 20.25 kHz. Scanning velocities of the laser spot over the sample of up to 200 mm/s were tested without a decline in perforation efficiency. This velocity leads to a process speed of ∼8 s per well of a 96 well plate. The optimal particle density was determined to be ∼6 particles per cell using environmental scanning electron microscopy. Applying the optimized parameters transfection efficiencies of 88% were achieved in canine pleomorphic adenoma ZMTH3 cells using a fluorescent labeled siRNA while maintaining a high cell viability of >90%. Gene knock down of d2-EGFP was demonstrated and validated by fluorescence repression and western blot analysis. On basis of our findings and established mathematical models we suppose a mixed transfection mechanism consisting of thermal and multiphoton near field effects. Our findings emphasize that gold nanoparticle mediated laser transfection provides an excellent tool for molecular delivery for both, high throughput purposes and the transfection of sensitive cells types.

  8. Gold Nanoparticle Mediated Laser Transfection for Efficient siRNA Mediated Gene Knock Down

    Science.gov (United States)

    Heinemann, Dag; Schomaker, Markus; Kalies, Stefan; Schieck, Maximilian; Carlson, Regina; Escobar, Hugo Murua; Ripken, Tammo; Meyer, Heiko; Heisterkamp, Alexander

    2013-01-01

    Laser based transfection methods have proven to be an efficient and gentle alternative to established molecule delivery methods like lipofection or electroporation. Among the laser based methods, gold nanoparticle mediated laser transfection bears the major advantage of high throughput and easy usability. This approach uses plasmon resonances on gold nanoparticles unspecifically attached to the cell membrane to evoke transient and spatially defined cell membrane permeabilization. In this study, we explore the parameter regime for gold nanoparticle mediated laser transfection for the delivery of molecules into cell lines and prove its suitability for siRNA mediated gene knock down. The developed setup allows easy usage and safe laser operation in a normal lab environment. We applied a 532 nm Nd:YAG microchip laser emitting 850 ps pulses at a repetition rate of 20.25 kHz. Scanning velocities of the laser spot over the sample of up to 200 mm/s were tested without a decline in perforation efficiency. This velocity leads to a process speed of ∼8 s per well of a 96 well plate. The optimal particle density was determined to be ∼6 particles per cell using environmental scanning electron microscopy. Applying the optimized parameters transfection efficiencies of 88% were achieved in canine pleomorphic adenoma ZMTH3 cells using a fluorescent labeled siRNA while maintaining a high cell viability of >90%. Gene knock down of d2-EGFP was demonstrated and validated by fluorescence repression and western blot analysis. On basis of our findings and established mathematical models we suppose a mixed transfection mechanism consisting of thermal and multiphoton near field effects. Our findings emphasize that gold nanoparticle mediated laser transfection provides an excellent tool for molecular delivery for both, high throughput purposes and the transfection of sensitive cells types. PMID:23536802

  9. Acceleration of gene transfection efficiency in neuroblastoma cells through polyethyleneimine/poly(methyl methacrylate) core-shell magnetic nanoparticles

    Science.gov (United States)

    Tencomnao, Tewin; Klangthong, Kewalin; Pimpha, Nuttaporn; Chaleawlert-umpon, Saowaluk; Saesoo, Somsak; Woramongkolchai, Noppawan; Saengkrit, Nattika

    2012-01-01

    Background The purpose of this study was to demonstrate the potential of magnetic poly(methyl methacrylate) (PMMA) core/polyethyleneimine (PEI) shell (mag-PEI) nanoparticles, which possess high saturation magnetization for gene delivery. By using mag-PEI nanoparticles as a gene carrier, this study focused on evaluation of transfection efficiency under magnetic induction. The potential role of this newly synthesized nanosphere for therapeutic delivery of the tryptophan hydroxylase-2 (TPH-2) gene was also investigated in cultured neuronal LAN-5 cells. Methods The mag-PEI nanoparticles were prepared by one-step emulsifier-free emulsion polymerization, generating highly loaded and monodispersed magnetic polymeric nanoparticles bearing an amine group. The physicochemical properties of the mag-PEI nanoparticles and DNA-bound mag-PEI nanoparticles were investigated using the gel retardation assay, atomic force microscopy, and zeta size measurements. The gene transfection efficiencies of mag-PEI nanoparticles were evaluated at different transfection times. Confocal laser scanning microscopy confirmed intracellular uptake of the magnetoplex. The optimal conditions for transfection of TPH-2 were selected for therapeutic gene transfection. We isolated the TPH-2 gene from the total RNA of the human medulla oblongata and cloned it into an expression vector. The plasmid containing TPH-2 was subsequently bound onto the surfaces of the mag-PEI nanoparticles via electrostatic interaction. Finally, the mag-PEI nanoparticle magnetoplex was delivered into LAN-5 cells. Reverse-transcriptase polymerase chain reaction was performed to evaluate TPH-2 expression in a quantitative manner. Results The study demonstrated the role of newly synthesized high-magnetization mag-PEI nanoparticles for gene transfection in vitro. The expression signals of a model gene, luciferase, and a therapeutic gene, TPH-2, were enhanced under magnetic-assisted transfection. An in vitro study in neuronal cells

  10. Efficient transfection of MG-63 osteoblasts using magnetic nanoparticles and oscillating magnetic fields.

    Science.gov (United States)

    Fouriki, A; Clements, M A; Farrow, N; Dobson, J

    2014-03-01

    To examine the potential of magnetic nanoparticles (MNPs) in transfecting human osteosarcoma fibroblasts (MG-63) and investigate the effects of a novel non-viral oscillating nanomagnetic gene transfection system (magnefect-nano™) in enhancing transfection efficiency (TE). MG-63 cells were transfected using MNPs coupled with a GFP-carrying plasmid. The magnefect-nano system was evaluated for transfection efficiency and potential associated effects on cell viability. MG-63 cells were efficiently transfected using MNPs and the magnefect-nano system significantly enhanced overall transfection efficiency. MNPs were not found to affect cell viability and/or function of the cells. Non-viral transfection using MNPs and the magnefect-nano system can be used to transfect MG-63 cells and assist reporter gene delivery on a single cell basis, highlighting the wide potential of nanomagnetic gene transfection in gene therapy. Copyright © 2012 John Wiley & Sons, Ltd.

  11. Efficient Gene Knockdown in Mouse Oocytes through Peptide Nanoparticle-Mediated SiRNA Transfection.

    Directory of Open Access Journals (Sweden)

    Zhen Jin

    Full Text Available The use of mouse oocytes as a model for studying female meiosis is very important in reproductive medicine. Gene knockdown by specific small interfering RNA (siRNA is usually the first step in the study of the function of a target gene in mouse oocytes during in vitro maturation. Traditionally, the only way to introduce siRNA into mouse oocytes is through microinjection, which is certainly less efficient and strenuous than siRNA transfection in somatic cells. Recently, in research using somatic cells, peptide nanoparticle-mediated siRNA transfection has been gaining popularity over liposome nanoparticle-mediated methods because of its high efficiency, low toxicity, good stability, and strong serum compatibility. However, no researchers have yet tried transfecting siRNA into mouse oocytes because of the existence of the protective zona pellucida surrounding the oocyte membrane (vitelline membrane. We therefore tested whether peptide nanoparticles can introduce siRNA into mouse oocytes. In the present study, we showed for the first time that our optimized program can efficiently knock down a target gene with high specificity. Furthermore, we achieved the expected meiotic phenotypes after we knocked down a test unknown target gene TRIM75. We propose that peptide nanoparticles may be superior for preliminary functional studies of unknown genes in mouse oocytes.

  12. Non-viral bone morphogenetic protein 2 transfection of rat dental pulp stem cells using calcium phosphate nanoparticles as carriers.

    Science.gov (United States)

    Yang, Xuechao; Walboomers, X Frank; van den Dolder, Juliette; Yang, Fang; Bian, Zhuan; Fan, Mingwen; Jansen, John A

    2008-01-01

    Calcium phosphate nanoparticles have shown potential as non-viral vectors for gene delivery. The aim of this study was to induce bone morphogenetic protein (Bmp)2 transfection in rat dental pulp stem cells using calcium phosphate nanoparticles as a gene vector and then to evaluate the efficiency and bioactivity of the transfection. We also intended to investigate the behavior of transfected cells when seeded on 3-dimensional titanium fiber mesh scaffolds. Nanoparticles of calcium phosphate encapsulating plasmid deoxyribonucleic acid (DNA) (plasmid enhanced green fluorescent protein-BMP2) were prepared. Then, STRO-1-selected rat dental pulp stem cells were transfected using these nanoparticles. Transfection and bioactivity of the secreted BMP2 were examined. Thereafter, the transfected cells were cultured on a fibrous titanium mesh. The cultures were investigated using scanning electron microscipy and evaluated for cell proliferation, alkaline phosphatase activity and calcium content. Finally, real-time polymerase chain reaction was performed for odontogenesis-related gene expression. The results showed that the size of the DNA-loaded particles was approximately 100 nm in diameter. Nanoparticles could protect the DNA encapsulated inside from external DNase and release the loaded DNA in a low-acid environment (pH 3.0). In vitro, nanoparticle transfection was shown to be effective and to accelerate or promote the odontogenic differentiation of rat dental pulp stem cells when cultured in the 3-dimensional scaffolds. Based on our results, plasmid DNA-loaded calcium phosphate nanoparticles appear to be an effective non-viral vector for gene delivery and functioned well for odontogenic differentiation through Bmp2 transfection.

  13. Non-viral bone morphogenetic protein 2 transfection of rat dental pulp stem cells using calcium phosphate nanoparticles as carriers.

    NARCIS (Netherlands)

    Yang, X.; Walboomers, X.F.; Dolder, J. van den; Yang, F.; Bian, Z.; Fan, M.; Jansen, J.A.

    2008-01-01

    Calcium phosphate nanoparticles have shown potential as non-viral vectors for gene delivery. The aim of this study was to induce bone morphogenetic protein (Bmp)2 transfection in rat dental pulp stem cells using calcium phosphate nanoparticles as a gene vector and then to evaluate the efficiency and

  14. Covalently bound DNA on naked iron oxide nanoparticles: Intelligent colloidal nano-vector for cell transfection.

    Science.gov (United States)

    Magro, Massimiliano; Martinello, Tiziana; Bonaiuto, Emanuela; Gomiero, Chiara; Baratella, Davide; Zoppellaro, Giorgio; Cozza, Giorgio; Patruno, Marco; Zboril, Radek; Vianello, Fabio

    2017-11-01

    Conversely to common coated iron oxide nanoparticles, novel naked surface active maghemite nanoparticles (SAMNs) can covalently bind DNA. Plasmid (pDNA) harboring the coding gene for GFP was directly chemisorbed onto SAMNs, leading to a novel DNA nanovector (SAMN@pDNA). The spontaneous internalization of SAMN@pDNA into cells was compared with an extensively studied fluorescent SAMN derivative (SAMN@RITC). Moreover, the transfection efficiency of SAMN@pDNA was evaluated and explained by computational model. SAMN@pDNA was prepared and characterized by spectroscopic and computational methods, and molecular dynamic simulation. The size and hydrodynamic properties of SAMN@pDNA and SAMN@RITC were studied by electron transmission microscopy, light scattering and zeta-potential. The two nanomaterials were tested by confocal scanning microscopy on equine peripheral blood-derived mesenchymal stem cells (ePB-MSCs) and GFP expression by SAMN@pDNA was determined. Nanomaterials characterized by similar hydrodynamic properties were successfully internalized and stored into mesenchymal stem cells. Transfection by SAMN@pDNA occurred and GFP expression was higher than lipofectamine procedure, even in the absence of an external magnetic field. A computational model clarified that transfection efficiency can be ascribed to DNA availability inside cells. Direct covalent binding of DNA on naked magnetic nanoparticles led to an extremely robust gene delivery tool. Hydrodynamic and chemical-physical properties of SAMN@pDNA were responsible of the successful uptake by cells and of the efficiency of GFP gene transfection. SAMNs are characterized by colloidal stability, excellent cell uptake, persistence in the host cells, low toxicity and are proposed as novel intelligent DNA nanovectors for efficient cell transfection. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Mannosylated Chitosan Nanoparticles Based Macrophage-Targeting Gene Delivery System Enhanced Cellular Uptake and Improved Transfection Efficiency.

    Science.gov (United States)

    Peng, Yixing; Yao, Wenjun; Wang, Bo; Zong, Li

    2015-04-01

    Gene transfer mediated by mannosylated chitosan (MCS) is a safe and promising approach for gene and vaccine delivery. MCS nanoparticles based gene delivery system showed high in vivo delivery efficiency and elicited strong immune responses in mice. However, little knowledge about the cell binding, transfection efficiency and intracellular trafficking of MCS nanoparticles had been acquired. In this study, using gastrin-releasing peptide as a model plasmid (pGRP), the binding of MCS/pGRP nanoparticles to macrophages and the intracellular trafficking of MCS/pGRP nanoparticles in macrophages were investigated. MCS-mediated transfection efficiency in macrophages was also evaluated using pGL-3 as a reporter gene. The results showed that the binding and transfection efficiency of MCS nanoparticles in macrophages was higher than that of CS, which was attributed to the interaction between mannose ligands in MCS and mannose receptors on the surface of macrophages. Observation with a confocal laser scanning microscope indicated the cellular uptake of MCS/pGRP nanoparticles were more than that of CS/pGRP nanoparticles in macrophages. MCS/pGRP nanoparticles were taken up by macrophages and most of them were entrapped in endosomal/lysosomal compartments. After the nanoparticles escaping from endosomal/lysosomal compartments, naked pGRP entered the nucleus, and a few MCS might enter the nucleus in terms of nanoparticles. Overall, MCS has the potential to be an excellent macrophage-targeting gene delivery carrier.

  16. Influence of nanoparticle-mediated transfection on proliferation of primary immune cells in vitro and in vivo.

    Science.gov (United States)

    Przybylski, Susanne; Gasch, Michaela; Marschner, Anne; Ebert, Marcus; Ewe, Alexander; Helmig, Gisa; Hilger, Nadja; Fricke, Stephan; Rudzok, Susanne; Aigner, Achim; Burkhardt, Jana

    2017-01-01

    One of the main obstacles in the widespread application of gene therapeutic approaches is the necessity for efficient and safe transfection methods. For the introduction of small oligonucleotide gene therapeutics into a target cell, nanoparticle-based methods have been shown to be highly effective and safe. While immune cells are a most interesting target for gene therapy, transfection might influence basic immune functions such as cytokine expression and proliferation, and thus positively or negatively affect therapeutic intervention. Therefore, we investigated the effects of nanoparticle-mediated transfection such as polyethylenimine (PEI) or magnetic beads on immune cell proliferation. Human adherent and non-adherent PBMCs were transfected by various methods (e.g. PEI, Lipofectamine® 2000, magnetofection) and stimulated. Proliferation was measured by lymphocyte transformation test (LTT). Cell cycle stages as well as expression of proliferation relevant genes were analyzed. Additionally, the impact of nanoparticles was investigated in vivo in a murine model of the severe systemic immune disease GvHD (graft versus host disease). The proliferation of primary immune cells was influenced by nanoparticle-mediated transfection. In particular in the case of magnetic beads, proliferation inhibition coincided with short-term cell cycle arrest and reduced expression of genes relevant for immune cell proliferation. Notably, proliferation inhibition translated into beneficial effects in a murine GvHD model with animals treated with PEI-nanoparticles showing increased survival (pPEI = 0.002) most likely due to reduced inflammation. This study shows for the first time that nanoparticles utilized for gene therapeutic transfection are able to alter proliferation of immune cells and that this effect depends on the type of nanoparticle. For magnetic beads, this was accompanied by temporary cell cycle arrest. Notably, in GvHD this nonspecific anti-proliferative effect might

  17. Effects of AC/DC magnetic fields, frequency, and nanoparticle aspect ratio on cellular transfection of gene vectors

    Science.gov (United States)

    Ford, Kris; Mair, Lamar; Fisher, Mike; Rowshon Alam, Md.; Juliano, Rudolph; Superfine, Richard

    2008-10-01

    In order to make non-viral gene delivery a useful tool in the study and treatment of genetic disorders, it is imperative that these methodologies be further refined to yield optimal results. Transfection of magnetic nanoparticles and nanorods are used as non-viral gene vectors to transfect HeLa EGFP-654 cells that stably express a mutated enhanced green fluorescent protein (EGFP) gene. We deliver antisense oligonucleotides to these cells designed to correct the aberrant splicing caused by the mutation in the EGFP gene. We also transfect human bronchial endothelial cells and immortalized WI-38 lung cells with pEGFP-N1 vectors. To achieve this we bind the genes to magnetic nanoparticles and nanorods and introduce magnetic fields to effect transfection. We wish to examine the effects of magnetic fields on the transfection of these particles and the benefits of using alternating (AC) magnetic fields in improving transfection rates over direct (DC) magnetic fields. We specifically look at the frequency dependence of the AC field and particle aspect ratio as it pertains to influencing transfection rate. We posit that the increase in angular momentum brought about by the AC field and the high aspect ratio of the nanorod particles, is vital to generating the force needed to move the particle through the cell membrane.

  18. In vitro studies of magnetically enhanced transfection in COS-7 cells

    International Nuclear Information System (INIS)

    Ang, D.; Tay, C.Y.; Tan, L.P.; Preiser, P.R.; Ramanujan, R.V.

    2011-01-01

    In the magnetically enhanced gene delivery technique, DNA complexed with polymer coated aggregated magnetic nanoparticles (AMNPs) is used for effecting transfection. The aim of this study is to examine the relationship between transfection efficiency and the physical characteristics of the polymer coated AMNPs. In vitro studies of transfection efficiency in COS-7 cells were carried out using pEGFP-N1 and pMIR-REPORT complexed polyethylenimine (PEI) coated iron oxide magnetic nanoparticles. PEI coated AMNPs (PEI-AMNPs) with average individual particle diameters in the range of 8 nm to 30 nm were studied and characterized by transmission electron microscopy, vibrating sample magnetometry, X-ray diffractometry, thermal gravimetric analysis and photon correlation spectroscopy methods. PEI-A8MNP and PEI-A30MNP yielded higher transfection efficiency compared to commercial polyMAG particles as well as PEI of equivalent molar ratio of nitrogen/phosphorous (N/P ratio). The transfection efficiency was related to the physical characteristics of the PEI-AMNPs and its complexes: transfection efficiency was strongly positively correlated with saturation magnetization (Ms) and susceptibility (χ), strongly negatively correlated with N/P ratio, moderately positively correlated to zeta potential and moderately negatively correlated to hydrodynamic diameter of the complex. PEI-A8MNP and PEI-A30MNP possessing higher Ms, χ, lower N/P ratio and smaller complex size exhibited higher transfection efficiency compared to PEI-A16MNP which have weaker magnetic properties, higher N/P ratio and larger complex size. We have demonstrated that optimization of the physical properties of PEI-AMNPs is needed to maximize transfection efficiency. - Research highlights: →The transfection efficiency in magnetically enhanced gene delivery was studied. →Transfection efficiency was strongly positively correlated to magnetic properties. →Transfection efficiency was strongly negatively correlated with

  19. Ocular nanoparticle toxicity and transfection of the retina and retinal pigment epithelium.

    Science.gov (United States)

    Prow, Tarl W; Bhutto, Imran; Kim, Sahng Y; Grebe, Rhonda; Merges, Carol; McLeod, D Scott; Uno, Koichi; Mennon, Mohamed; Rodriguez, Li; Leong, Kam; Lutty, Gerard A

    2008-12-01

    Chitosan, PCEP (poly{[(cholesteryl oxocarbonylamido ethyl) methyl bis(ethylene) ammonium iodide] ethyl phosphate}), and magnetic nanoparticles (MNPs) were evaluated for the safe delivery of genes in the eye. Rabbits were injected with nanoparticles either intravitreally (IV) or subretinally (SR) and sacrificed 7 days later. Eyes were grossly evaluated for retinal pigment epithelium abnormalities, retinal degeneration, and inflammation. All eyes were cryopreserved and sectioned for analysis of toxicity and expression of either enhanced green or red fluorescent proteins. All of the nanoparticles were able to transfect cells in vitro and in vivo. IV chitosan showed inflammation in 12/13 eyes, whereas IV PCEP and IV MNPs were not inflammatory and did not induce retinal pathology. SR PCEP was nontoxic in the majority of cases but yielded poor transfection, whereas SR MNPs were nontoxic and yielded good transfection. Therefore, we conclude that the best nanoparticle evaluated in vivo was the least toxic nanoparticle tested, the MNP.

  20. A general strategy to achieve ultra-high gene transfection efficiency using lipid-nanoparticle composites.

    Science.gov (United States)

    Vankayala, Raviraj; Chiang, Chi-Shiun; Chao, Jui-I; Yuan, Chiun-Jye; Lin, Shyr-Yeu; Hwang, Kuo Chu

    2014-09-01

    Gene therapy provides a new hope for previously "incurable" diseases. Low gene transfection efficiency, however, is the bottle-neck to the success of gene therapy. It is very challenging to develop non-viral nanocarriers to achieve ultra-high gene transfection efficiencies. Herein, we report a novel design of "tight binding-but-detachable" lipid-nanoparticle composite to achieve ultrahigh gene transfection efficiencies of 60∼82%, approaching the best value (∼90%) obtained using viral vectors. We show that Fe@CNPs nanoparticles coated with LP-2000 lipid molecules can be used as gene carriers to achieve ultra-high (60-80%) gene transfection efficiencies in HeLa, U-87MG, and TRAMP-C1 cells. In contrast, Fe@CNPs having surface-covalently bound N,N,N-trimethyl-N-2-methacryloxyethyl ammonium chloride (TMAEA) oligomers can only achieve low (23-28%) gene transfection efficiencies. Similarly ultrahigh gene transfection/expression was also observed in zebrafish model using lipid-coated Fe@CNPs as gene carriers. Evidences for tight binding and detachability of DNA from lipid-nanoparticle nanocarriers will be presented. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Poly(β-Amino Ester)-Nanoparticle Mediated Transfection of Retinal Pigment Epithelial Cells In Vitro and In Vivo

    Science.gov (United States)

    Bhutto, Imran; Handa, James T.; Green, Jordan J.

    2012-01-01

    A variety of genetic diseases in the retina, including retinitis pigmentosa and leber congenital amaurosis, might be excellent targets for gene delivery as treatment. A major challenge in non-viral gene delivery remains finding a safe and effective delivery system. Poly(beta-amino ester)s (PBAEs) have shown great potential as gene delivery reagents because they are easily synthesized and they transfect a wide variety of cell types with high efficacy in vitro. We synthesized a combinatorial library of PBAEs and evaluated them for transfection efficacy and toxicity in retinal pigment epithelial (ARPE-19) cells to identify lead polymer structures and transfection formulations. Our optimal polymer (B5-S5-E7 at 60 w/w polymer∶DNA ratio) transfected ARPE-19 cells with 44±5% transfection efficacy, significantly higher than with optimized formulations of leading commercially available reagents Lipofectamine 2000 (26±7%) and X-tremeGENE HP DNA (22±6%); (ptransfection efficacy. This high non-viral efficacy was achieved with comparable cytotoxicity (23±6%) to controls; optimized formulations of Lipofectamine 2000 and X-tremeGENE HP DNA showed 15±3% and 32±9% toxicity respectively (p>0.05 for both). Our optimal polymer was also significantly better than a gold standard polymeric transfection reagent, branched 25 kDa polyethyleneimine (PEI), which achieved only 8±1% transfection efficacy with 25±6% cytotoxicity. Subretinal injections using lyophilized GFP-PBAE nanoparticles resulted in 1.1±1×103-fold and 1.5±0.7×103-fold increased GFP expression in the retinal pigment epithelium (RPE)/choroid and neural retina respectively, compared to injection of DNA alone (p = 0.003 for RPE/choroid, ptransfection of the RPE in vivo suggests that these nanoparticles could be used to study a number of genetic diseases in the laboratory with the potential to treat debilitating eye diseases. PMID:22629417

  2. Delivery of proteins to mammalian cells via gold nanoparticle mediated laser transfection

    International Nuclear Information System (INIS)

    Heinemann, D; Kalies, S; Schomaker, M; Ertmer, W; Meyer, H; Ripken, T; Murua Escobar, H

    2014-01-01

    Nanoparticle laser interactions are in widespread use in cell manipulation. In particular, molecular medicine needs techniques for the directed delivery of molecules into mammalian cells. Proteins are the final mediator of most cellular cascades. However, despite several methodical approaches, the efficient delivery of proteins to cells remains challenging. This paper presents a new protein transfection technique via laser scanning of cells previously incubated with gold nanoparticles. The laser-induced plasmonic effects on the gold nanoparticles cause a transient permeabilization of the cellular membrane, allowing proteins to enter the cell. Applying this technique, it was possible to deliver green fluorescent protein into mammalian cells with an efficiency of 43%, maintaining a high level of cell viability. Furthermore, a functional delivery of Caspase 3, an apoptosis mediating protein, was demonstrated and evaluated in several cellular assays. Compared to conventional protein transfection techniques such as microinjection, the methodical approach presented here enables high-throughput transfection of about 10 000 cells per second. Moreover, a well-defined point in time of delivery is guaranteed by gold nanoparticle mediated laser transfection, allowing the detailed temporal analysis of cellular pathways and protein trafficking. (papers)

  3. Cationic solid-lipid nanoparticles can efficiently bind and transfect plasmid DNA

    NARCIS (Netherlands)

    Olbrich, C; Bakowsky, U; Muller, RH; Kneuer, C

    2001-01-01

    The suitability of cationically modified solid-lipid nanoparticles (SLN) as a novel transfection agent was investigated. SLN were produced by hot homogenisation using either Compritol ATO 888 or paraffin as matrix lipid, a mixture of Tween 80 and Span 85 as tenside and either EQ1

  4. Nonviral gene transfection nanoparticles: function and applications in the brain.

    Science.gov (United States)

    Roy, Indrajit; Stachowiak, Michal K; Bergey, Earl J

    2008-06-01

    In vivo transfer and expression of foreign genes allows for the elucidation of functions of genes in living organisms and generation of disease models in animals that more closely resemble the etiology of human diseases. Gene therapy holds promise for the cure of a number of diseases at the fundamental level. Synthetic "nonviral" materials are fast gaining popularity as safe and efficient vectors for delivering genes to target organs. Not only can nanoparticles function as efficient gene carriers, they also can simultaneously carry diagnostic probes for direct "real-time" visualization of gene transfer and downstream processes. This review has focused on the central nervous system (CNS) as the target for nonviral gene transfer, with special emphasis on organically modified silica (ORMOSIL) nanoparticles developed in our laboratory. These nanoparticles have shown robust gene transfer efficiency in brain cells in vivo and allowed to investigate mechanisms that control neurogenesis as well as neurodegenerative disorders.

  5. Transfection using hydroxyapatite nanoparticles in the inner ear via an intact round window membrane in chinchilla

    Energy Technology Data Exchange (ETDEWEB)

    Wu Xuewen; Ding Dalian [Central South University, Department of Otolaryngology Head and Neck Surgery, The Third Xiangya Hospital (China); Jiang Haiyan [State University of New York at Buffalo, Center for Hearing and Deafness (United States); XingXiaowei [Central South University, Department of Otolaryngology Head and Neck Surgery, The Third Xiangya Hospital (China); Huang, Suping [Central South University, State Key Laboratory of Powder Metallurgy (China); Liu Hong [Central South University, Department of Otolaryngology Head and Neck Surgery, The Third Xiangya Hospital (China); Chen Zhedong [Central South University, State Key Laboratory of Powder Metallurgy (China); Sun Hong, E-mail: shjhaj@vip.163.com [Central South University, Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital (China)

    2012-01-15

    Hydroxyapatite nanoparticles (nHAT) are known to have excellent biocompatibility, and have attracted increasing attention as new candidates of non-viral vectors for gene therapy. In our previous studies, nHAT carrying a therapeutic gene and a reporter gene were successfully transfected into the spiral ganglion neurons in the inner ear of guinea pigs in vivo as well as in the cultured cell lines, although the transfection efficiencies were never higher than 30%. In this study, the surface modification of nHAT with polyethylenimine (PEI) was made (PEI-nHAT, diameter = 73.09 {+-} 27.32 nm) and a recombinant plasmid carrying enhanced green fluorescent protein (EGFP) gene and neurotrophin-3 (NT-3) gene was constructed as pEGFPC2-NT3. The PEI modified nHAT and the recombinant plasmid was then connected to form the nHAT-based vector-gene complex (PEI-nHAT-pEGFPC2-NT3). This complex was then placed onto the intact round window membranes of the chinchillas for inner ear transfection. Auditory brainstem response (ABR) was tested to evaluate auditory function. Green fluorescence of EGFP was observed using confocal microscopy 48 h after administering vector-gene complexes. There was no significant threshold shift in tone burst-evoked ABR at any tested frequency. Abundant, condensed green fluorescence was found in dark cells on both sides of the crista and around the macula of the utricle. Scattered EGFP signals were also detected in vestibular hair cells, some Schwann cells in the cochlear spiral ganglion region, some outer pillar cells in the organ of Corti, and a few cells in the stria vascularis. The density of green fluorescence-marked cells was obviously higher in the vestibular dark cell area than in other areas of the inner ear, suggesting that vestibular dark cells may have the ability to actively engulf the nHAT-based vector-gene complexes. Considering the high transfection efficiency in the vestibular system, PEI-nHAT may be a potential vector for gene therapy of

  6. Transfection using hydroxyapatite nanoparticles in the inner ear via an intact round window membrane in chinchilla

    International Nuclear Information System (INIS)

    Wu Xuewen; Ding Dalian; Jiang Haiyan; XingXiaowei; Huang, Suping; Liu Hong; Chen Zhedong; Sun Hong

    2012-01-01

    Hydroxyapatite nanoparticles (nHAT) are known to have excellent biocompatibility, and have attracted increasing attention as new candidates of non-viral vectors for gene therapy. In our previous studies, nHAT carrying a therapeutic gene and a reporter gene were successfully transfected into the spiral ganglion neurons in the inner ear of guinea pigs in vivo as well as in the cultured cell lines, although the transfection efficiencies were never higher than 30%. In this study, the surface modification of nHAT with polyethylenimine (PEI) was made (PEI–nHAT, diameter = 73.09 ± 27.32 nm) and a recombinant plasmid carrying enhanced green fluorescent protein (EGFP) gene and neurotrophin-3 (NT-3) gene was constructed as pEGFPC2–NT3. The PEI modified nHAT and the recombinant plasmid was then connected to form the nHAT-based vector–gene complex (PEI–nHAT–pEGFPC2–NT3). This complex was then placed onto the intact round window membranes of the chinchillas for inner ear transfection. Auditory brainstem response (ABR) was tested to evaluate auditory function. Green fluorescence of EGFP was observed using confocal microscopy 48 h after administering vector–gene complexes. There was no significant threshold shift in tone burst-evoked ABR at any tested frequency. Abundant, condensed green fluorescence was found in dark cells on both sides of the crista and around the macula of the utricle. Scattered EGFP signals were also detected in vestibular hair cells, some Schwann cells in the cochlear spiral ganglion region, some outer pillar cells in the organ of Corti, and a few cells in the stria vascularis. The density of green fluorescence-marked cells was obviously higher in the vestibular dark cell area than in other areas of the inner ear, suggesting that vestibular dark cells may have the ability to actively engulf the nHAT-based vector–gene complexes. Considering the high transfection efficiency in the vestibular system, PEI–nHAT may be a potential vector for

  7. Purification of transfection-grade plasmid DNA from bacterial cells with superparamagnetic nanoparticles

    Science.gov (United States)

    Chiang, Chen-Li; Sung, Ching-Shan

    2006-07-01

    The functionalized magnetic nanobeads were used to develop a rapid protocol for extracting and purifying transfection-grade plasmid DNA from bacterial culture. Nanosized superparamagnetic nanoparticles (Fe 3O 4) were prepared by chemical coprecipitation method using Fe 2+, Fe 3+ salt, and ammonium hydroxide under a nitrogen atmosphere. The surface of Fe 3O 4 nanoparticles was modified by coating with the multivalent cationic agent, polyethylenimine (PEI). The PEI-modified magnetic nanobeads were employed to simplify the purification of plasmid DNA from bacterial cells. We demonstrated a useful plasmid, pRSETB-EGFP, encoding the green fluorescent protein with T7 promoter, was amplified in DE3 strain of Escherichia coli. The loaded nanobeads are recovered by magnetically driven separation and regenerated by exposure to the elution buffer with optimal ionic strength (1.25 M) and pH (9.0). Up to approximately 819 μg of high-purity (A 260/A 280 ratio=1.86) plasmid DNA was isolated from 100 ml of overnight bacterial culture. The eluted plasmid DNA was used directly for restriction enzyme digestion, bacterial cell transformation and animal cell transfection applications with success. The PEI-modified magnetic nanobead delivers significant time-savings, overall higher yields and better transfection efficiencies compared to anion-exchange and other methods. The results presented in this report show that PEI-modified magnetic nanobeads are suitable for isolation and purification of transfection-grade plasmid DNA.

  8. Live-cell imaging to compare the transfection and gene silencing efficiency of calcium phosphate nanoparticles and a liposomal transfection agent.

    Science.gov (United States)

    Chernousova, S; Epple, M

    2017-05-01

    The processing of DNA (for transfection) and short interfering RNA (siRNA; for gene silencing), introduced into HeLa cells by triple-shell calcium phosphate nanoparticles, was followed by live-cell imaging. For comparison, the commercial liposomal transfection agent Lipofectamine was used. The cells were incubated with these delivery systems, carrying either enhanced green fluorescent protein (eGFP)-encoding DNA or siRNA against eGFP. In the latter case, HeLa cells that stably expressed eGFP were used. The expression of eGFP started after 5 h in the case of nanoparticles and after 4 h in the case of Lipofectamine. The corresponding times for gene silencing were 5 h (nanoparticles) and immediately after incubation (Lipofectamine). The expression of eGFP was notably enhanced 2-3 h after cell division (mitosis). In general, the transfection and gene silencing efficiencies of the nanoparticles were lower than those of Lipofectamime, even at a substantially higher dose (factor 20) of nucleic acids. However, the cytotoxicity of the nanoparticles was lower than that of Lipofectamine, making them suitable vectors for in vivo application.

  9. Gene transfection of human mesenchymal stem cells with a nano-hydroxyapatite-collagen scaffold containing DNA-functionalized calcium phosphate nanoparticles.

    Science.gov (United States)

    Tenkumo, Taichi; Vanegas Sáenz, Juan Ramón; Takada, Yukyo; Takahashi, Masatoshi; Rotan, Olga; Sokolova, Viktoriya; Epple, Matthias; Sasaki, Keiichi

    2016-07-01

    This study aimed to fabricate a growth factor-releasing biodegradable scaffold for tissue regeneration. We prepared multishell calcium phosphate (CaP) nanoparticles functionalized with DNA, polyethyleneimine (PEI), protamine and octa-arginine (R8) and compared their respective transfection activity and cell viability measures using human mesenchymal stem cells. DNA-protamine complexes improved the transfection efficiency of CaP nanoparticles with the exception of those functionalized with R8. These complexes also greatly reduced the cytotoxicity of PEI. In addition, we also fabricated DNA-protamine-functionalized CaP nanoparticle-loaded nano-hydroxyapatite-collagen scaffolds and investigated their gene transfection efficiencies. These experiments showed that the scaffolds were associated with moderate hMSC cell viability and were capable of releasing the BMP-2 protein into hMSCs following gene transfection. In particular, the scaffold loaded with protamine-containing CaP nanoparticles showed the highest cell viability and transfection efficiency in hMSCs; thus, it might be suitable to serve as an efficient growth factor-releasing scaffold. © 2016 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

  10. Inclusion of the helper lipid dioleoyl-phosphatidylethanolamine in solid lipid nanoparticles inhibits their transfection efficiency.

    Science.gov (United States)

    de Jesus, Marcelo B; Radaic, Allan; Hinrichs, Wouter L J; Ferreira, Carmen V; de Paula, Eneida; Hoekstra, Dick; Zuhorn, Inge S

    2014-02-01

    Solid lipid nanoparticles (SLNs) are a promising system for the delivery of lipophilic and hydrophilic drugs. They consist of a solid lipid core that is stabilized by a layer of surfactants. By the incorporation of cationic lipids in the formulation, positively charged SLNs can be generated, that are suitable carriers for nucleic acids (DNA, siRNA). Considering the beneficial effect of helper lipids on the transfection efficiency with cationic liposomes, the effect of the helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) on transfection with cationic lipid-containing solid lipid nanoparticles was investigated in PC3 prostate cancer cells. The inclusion of DOPE in SLN formulations, instead of promoted, strongly inhibited SLN transfection efficiency, by frustrating the accommodation of DNA by the particles, as was revealed by biochemical analysis. SLNs devoid of DOPE maintained a homogenous size distribution of approximately 150 nm following lipoplex assembly and cellular delivery, and showed transfection efficiency comparable to that of Lipofectamine 2000' (LF2k). Moreover, the SLNs maintain their high transfection efficiency after lyophilization and long-term storage (1-2 years), an important asset for biomedical applications. There is even the possibility to lyophilize the SLN carrier together with its DNA cargo, which represents an interesting pharmaceutical advantage of the SLN formulations over LF2k. These results reflect marked differences between the physicochemical properties of cationic liposomes and SLNs, the latter requiring more critical lipid-depending properties for effective 'packaging' of DNA but displaying a higher storage stability than cationic lipid based carriers like LF2k.

  11. Comparison of chitosan, alginate and chitosan/alginate nanoparticles with respect to their size, stability, toxicity and transfection

    OpenAIRE

    Aras Rafiee; Mohammad Hossein Alimohammadian; TaranehGazori; Farhad Riazi-rad; Seyed Mohammad Reza Fatemi; Amirabbas Parizadeh; Ismaeil Haririan; Mohammad Havaskary

    2014-01-01

    Objective: To to compare the chitosan/alginate, chitosan and alginate nanoparticles as plasmid vectors, to determine the morphological characteristics, size and physicochemical properties of nanoparticle-pEGFP complexes and to evaluate the potential of these nanoparticles in transfection of pEGFP plasmid in to a cultured the human embryonic kidney cell line (HEK 293 cells). Methods: Nanoparticles comprising chitosan, alginate and both chitosan-alginate polymers were formed t...

  12. Cell transfection as a tool to study growth hormone action

    DEFF Research Database (Denmark)

    Norstedt, G; Enberg, B; Francis, S

    1994-01-01

    of cellular function that mimic those of the endogenous GHR. GHR cDNA transfected cells also offer a system where the mechanism of GH action can be studied. Such a system has been used to demonstrate that the GHR itself becomes tyrosine phosphorylated and that further phosphorylation of downstream proteins...... is important in GH action. The GH signals are transmitted to the nucleus and GH regulated genes have now begun to be characterized. The ability to use cell transfection for mechanistic studies of GH action will be instrumental to define domains within the receptor that are of functional importance...

  13. Characterization of nanoparticle mediated laser transfection by femtosecond laser pulses for applications in molecular medicine.

    Science.gov (United States)

    Schomaker, Markus; Heinemann, Dag; Kalies, Stefan; Willenbrock, Saskia; Wagner, Siegfried; Nolte, Ingo; Ripken, Tammo; Murua Escobar, Hugo; Meyer, Heiko; Heisterkamp, Alexander

    2015-02-03

    In molecular medicine, the manipulation of cells is prerequisite to evaluate genes as therapeutic targets or to transfect cells to develop cell therapeutic strategies. To achieve these purposes it is essential that given transfection techniques are capable of handling high cell numbers in reasonable time spans. To fulfill this demand, an alternative nanoparticle mediated laser transfection method is presented herein. The fs-laser excitation of cell-adhered gold nanoparticles evokes localized membrane permeabilization and enables an inflow of extracellular molecules into cells. The parameters for an efficient and gentle cell manipulation are evaluated in detail. Efficiencies of 90% with a cell viability of 93% were achieved for siRNA transfection. The proof for a molecular medical approach is demonstrated by highly efficient knock down of the oncogene HMGA2 in a rapidly proliferating prostate carcinoma in vitro model using siRNA. Additionally, investigations concerning the initial perforation mechanism are conducted. Next to theoretical simulations, the laser induced effects are experimentally investigated by spectrometric and microscopic analysis. The results indicate that near field effects are the initial mechanism of membrane permeabilization. This methodical approach combined with an automated setup, allows a high throughput targeting of several 100,000 cells within seconds, providing an excellent tool for in vitro applications in molecular medicine. NIR fs lasers are characterized by specific advantages when compared to lasers employing longer (ps/ns) pulses in the visible regime. The NIR fs pulses generate low thermal impact while allowing high penetration depths into tissue. Therefore fs lasers could be used for prospective in vivo applications.

  14. Plasmid transfection in mammalian cells spatiotemporally tracked by a gold nanoparticle.

    Science.gov (United States)

    Muroski, Megan E; Carnevale, Kate J F; Riskowski, Ryan A; Strouse, Geoffrey F

    2015-01-27

    Recent advances in cell transfection have suggested that delivery of a gene on a gold nanoparticle (AuNP) can enhance transfection efficiency. The mechanism of transfection is poorly understood, particularly when the gene is appended to a AuNP, as expression of the desired exogenous protein is dependent not only on the efficiency of the gene being taken into the cell but also on efficient endosomal escape and cellular processing of the nucleic acid. Design of a multicolor surface energy transfer (McSET) molecular beacon by independently dye labeling a linearized plasmid and short duplex DNA (sdDNA) appended to a AuNP allows spatiotemporal profiling of the transfection events, providing insight into package uptake, disassembly, and final plasmid expression. Delivery of the AuNP construct encapsulated in Lipofectamine2000 is monitored in Chinese hamster ovary cells using live-cell confocal microscopy. The McSET beacon signals the location and timing of the AuNP release and endosomal escape events for the plasmid and the sdDNA discretely, which are correlated with plasmid transcription by fluorescent protein expression within the cell. It is observed that delivery of the construct leads to endosomal release of the plasmid and sdDNA from the AuNP surface at different rates, prior to endosomal escape. Slow cytosolic diffusion of the nucleic acids is believed to be the limiting step for transfection, impacting the time-dependent expression of protein. The overall protein expression yield is enhanced when delivered on a AuNP, possibly due to better endosomal escape or lower degradation prior to endosomal escape.

  15. Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles

    Science.gov (United States)

    Tenkumo, Taichi; Kamano, Yuya; Egusa, Hiroshi; Sasaki, Keiichi

    2017-01-01

    Nanoparticles represent promising gene delivery systems in biomedicine to facilitate prolonged gene expression with low toxicity compared to viral vectors. Specifically, nanoparticles of calcium phosphate (nCaP), the main inorganic component of human bone, exhibit high biocompatibility and good biodegradability and have been reported to have high affinity for protein or DNA, having thus been used as gene transfer vectors. On the other hand, Octa-arginine (R8), which has a high permeability to cell membrane, has been reported to improve intracellular delivery systems. Here, we present an optimized method for nCaP-mediated gene delivery using an octa-arginine (R8)-functionalized nCaP vector containing a marker or functional gene construct. nCaP particle size was between 220–580 nm in diameter and all R8-functionalized nCaPs carried a positive charge. R8 concentration significantly improved nCaP transfection efficiency with high cell compatibility in human mesenchymal stem cells (hMSC) and human osteoblasts (hOB) in particular, suggesting nCaPs as a good option for non-viral vector gene delivery. Furthermore, pre-treatment with different endocytosis inhibitors identified that the endocytic pathway differed among cell lines and functionalized nanoparticles, with amiloride increasing transfection efficiency of R8-functionalized nCaPs in hMSC and hOB. PMID:29145481

  16. Comparison of chitosan, alginate and chitosan/alginate nanoparticles with respect to their size, stability, toxicity and transfection

    Directory of Open Access Journals (Sweden)

    Aras Rafiee

    2014-10-01

    Full Text Available Objective: To to compare the chitosan/alginate, chitosan and alginate nanoparticles as plasmid vectors, to determine the morphological characteristics, size and physicochemical properties of nanoparticle-pEGFP complexes and to evaluate the potential of these nanoparticles in transfection of pEGFP plasmid in to a cultured the human embryonic kidney cell line (HEK 293 cells. Methods: Nanoparticles comprising chitosan, alginate and both chitosan-alginate polymers were formed through pregel preparation method. The ability of plasmid-complexes in preventing DNA migration were assessed by the agarose gel assay. The efficiency of nanoparticles in transfection of pEGFP plasmid in the cultured HEK 293 cells was measured by flow cytometry. The effect of the nanoparticle-plasmid complexes on the cell viability was determined using cytotoxicity assay. Results: Chitosan, alginate and alginate/chitosan nanoparticles had a mean Z-average diameter of 620 nm, 235.8 nm and 161.8 nm and mean zeta potential of 45 mV, -18.6 mV and 29.3 mV, respectively. Chitosan and chitosan/alginate nanoparticles have greater capacity to maintain plasmid than alginate nanoparticles. Alginate nanoparticles had the greater transfection in comparison to the others. Cell viability assays indicated that nanoparticles had no toxic effect on HEK 293 cells after 4 h or 24 h. Conclusions: The combination of particle surface, hydrophobicity size and zeta potential can influence on transfection efficiency and the cellular uptake of the nanoparticles. Our suitable candidate for gene delivery would be alginate/chitosan nanoparticles.

  17. Transfection of Primary Human Skin Fibroblasts for Peroxisomal Studies

    NARCIS (Netherlands)

    Koster, Janet; Waterham, Hans R.

    2017-01-01

    Functional studies with primary human skin fibroblasts from patients with a peroxisomal disorder often require efficient transfection with plasmids to correct the genetic defect or to express heterologous reporter proteins. Here, we describe a protocol we commonly use for efficient nonviral

  18. Structural mediation on polycation nanoparticles by sulfadiazine to enhance DNA transfection efficiency and reduce toxicity.

    Science.gov (United States)

    Long, Xingwen; Zhang, Zhihui; Han, Shangcong; Tang, Minjie; Zhou, Junhui; Zhang, Jianhua; Xue, Zhenyi; Li, Yan; Zhang, Rongxin; Deng, Liandong; Dong, Anjie

    2015-04-15

    Reducing the toxicity while maintaining high transfection efficiency is an important issue for cationic polymers as gene carriers in clinical application. In this paper, a new zwitterionic copolymer, polycaprolactone-g-poly(dimethylaminoethyl methyacrylate-co-sulfadiazine methacrylate) (PC-SDZ) with unique pH-sensitivity, was designed and prepared. The incorporation of sulfadiazine into poly(dimethylaminoethyl methacrylate) (PDMAEMA) chains successfully mediates the surface properties including compacter shell structure, lower density of positive charges, stronger proton buffer capability, and enhanced hydrophobicity, which lead to reduction in toxicity and enhancements in stability, cellular uptake, endosome escape, and transfection efficiency for the PC-SDZ2 nanoparticles (NPs)/DNA complexes. Excellent transfection efficiency at the optimal N/P ratio of 10 was observed for PC-SDZ2 NPs/DNA complexes, which was higher than that of the commercial reagent-branched polyethylenimine (PEI). The cytotoxicity was evaluated by CCK8 measurement, and the results showed significant reduction in cytotoxicity even at high concentration of complexes after sulfadiazine modification. Therefore, this work may demonstrate a new way of structural mediation of cationic polymer carriers for gene delivery with high efficiency and low toxicity.

  19. Enhancing oligodendrocyte differentiation by transient transcription activation via DNA nanoparticle-mediated transfection.

    Science.gov (United States)

    Li, Xiaowei; Tzeng, Stephany Y; Zamboni, Camila Gadens; Koliatsos, Vassilis E; Ming, Guo-Li; Green, Jordan J; Mao, Hai-Quan

    2017-05-01

    Current approaches to derive oligodendrocytes from human pluripotent stem cells (hPSCs) need extended exposure of hPSCs to growth factors and small molecules, which limits their clinical application because of the lengthy culture time required and low generation efficiency of myelinating oligodendrocytes. Compared to extrinsic growth factors and molecules, oligodendrocyte differentiation and maturation can be more effectively modulated by regulation of the cell transcription network. In the developing central nervous system (CNS), two basic helix-loop-helix transcription factors, Olig1 and Olig2, are decisive in oligodendrocyte differentiation and maturation. Olig2 plays a critical role in the specification of oligodendrocytes and Olig1 is crucial in promoting oligodendrocyte maturation. Recently viral vectors have been used to overexpress Olig2 and Olig1 in neural stem/progenitor cells (NSCs) to induce the maturation of oligodendrocytes and enhance the remyelination activity in vivo. Because of the safety issues with viral vectors, including the insertional mutagenesis and potential tumor formation, non-viral transfection methods are preferred for clinical translation. Here we report a poly(β-amino ester) (PBAE)-based nanoparticle transfection method to deliver Olig1 and Olig2 into human fetal tissue-derived NSCs and demonstrate efficient oligodendrocyte differentiation following transgene expression of Olig1 and Olig2. This approach is potentially translatable for engineering stem cells to treat injured or diseased CNS tissues. Current protocols to derive oligodendrocytes from human pluripotent stem cells (hPSCs) require lengthy culture time with low generation efficiencies of mature oligodendrocytes. We described a new approach to enhance oligodendrocyte differentiation through nanoparticle-mediated transcription modulation. We tested an effective transfection method using cell-compatible poly (β-amino ester) (PBAE)/DNA nanoparticles as gene carrier to deliver

  20. Cationic nanoparticles with quaternary ammonium-functionalized PLGA-PEG-based copolymers for potent gene transfection

    Science.gov (United States)

    Wang, Yan-Hsung; Fu, Yin-Chih; Chiu, Hui-Chi; Wang, Chau-Zen; Lo, Shao-Ping; Ho, Mei-Ling; Liu, Po-Len; Wang, Chih-Kuang

    2013-11-01

    The objective of the present work was to develop new cationic nanoparticles (cNPs) with amphiphilic cationic copolymers for the delivery of plasmid DNA (pDNA). Cationic copolymers were built on the synthesis of quaternary ammonium salt compounds from diethylenetriamine (DETA) to include the positively charged head group and amphiphilic multi-grafts. PLGA- phe-PEG- qDETA (PPD), phe-PEG- qDETA-PLGA (PDP), and PLGA- phe-PEG- qDETA-PLGA (PPDP) cationic copolymers were created by this moiety of DETA quaternary ammonium, heterobifunctional polyethylene glycol (COOH-PEG-NH2), phenylalanine ( phe), and poly(lactic- co-glycolic acid) (PLGA). These new cNPs were prepared by the water miscible solvent displacement method. They exhibit good pDNA binding ability, as shown in a retardation assay that occurred at a particle size of 217 nm. The zeta potential was approximately +21 mV when the cNP concentration was 25 mg/ml. The new cNPs also have a better buffering capacity than PLGA NPs. However, the pDNA binding ability was demonstrated starting at a weight ratio of approximately 6.25 cNPs/pDNA. Gene transfection results showed that these cNPs had transfection effects similar to those of Lipofectamine 2000 in 293T cells. Furthermore, cNPs can also transfect human adipose-derived stem cells. The results indicate that the newly developed cNP is a promising candidate for a novel gene delivery vehicle.

  1. Cationic nanoparticles with quaternary ammonium-functionalized PLGA–PEG-based copolymers for potent gene transfection

    International Nuclear Information System (INIS)

    Wang, Yan-Hsung; Fu, Yin-Chih; Chiu, Hui-Chi; Wang, Chau-Zen; Lo, Shao-Ping; Ho, Mei-Ling; Liu, Po-Len; Wang, Chih-Kuang

    2013-01-01

    The objective of the present work was to develop new cationic nanoparticles (cNPs) with amphiphilic cationic copolymers for the delivery of plasmid DNA (pDNA). Cationic copolymers were built on the synthesis of quaternary ammonium salt compounds from diethylenetriamine (DETA) to include the positively charged head group and amphiphilic multi-grafts. PLGA-phe-PEG-qDETA (PPD), phe-PEG-qDETA-PLGA (PDP), and PLGA-phe-PEG-qDETA-PLGA (PPDP) cationic copolymers were created by this moiety of DETA quaternary ammonium, heterobifunctional polyethylene glycol (COOH-PEG-NH 2 ), phenylalanine (phe), and poly(lactic-co-glycolic acid) (PLGA). These new cNPs were prepared by the water miscible solvent displacement method. They exhibit good pDNA binding ability, as shown in a retardation assay that occurred at a particle size of ∼217 nm. The zeta potential was approximately +21 mV when the cNP concentration was 25 mg/ml. The new cNPs also have a better buffering capacity than PLGA NPs. However, the pDNA binding ability was demonstrated starting at a weight ratio of approximately 6.25 cNPs/pDNA. Gene transfection results showed that these cNPs had transfection effects similar to those of Lipofectamine 2000 in 293T cells. Furthermore, cNPs can also transfect human adipose-derived stem cells. The results indicate that the newly developed cNP is a promising candidate for a novel gene delivery vehicle

  2. A magnetic nanoparticle-based multiple-gene delivery system for transfection of porcine kidney cells.

    Directory of Open Access Journals (Sweden)

    Yan Wang

    Full Text Available Superparamagnetic nanoparticles are promising candidates for gene delivery into mammalian somatic cells and may be useful for reproductive cloning using the somatic cell nuclear transfer technique. However, limited investigations of their potential applications in animal genetics and breeding, particularly multiple-gene delivery by magnetofection, have been performed. Here, we developed a stable, targetable and convenient system for delivering multiple genes into the nuclei of porcine somatic cells using magnetic Fe3O4 nanoparticles as gene carriers. After surface modification by polyethylenimine, the spherical magnetic Fe3O4 nanoparticles showed strong binding affinity for DNA plasmids expressing the genes encoding a green (DNAGFP or red (DNADsRed fluorescent protein. At weight ratios of DNAGFP or DNADsRed to magnetic nanoparticles lower than or equal to 10∶1 or 5∶1, respectively, the DNA molecules were completely bound by the magnetic nanoparticles. Atomic force microscopy analyses confirmed binding of the spherical magnetic nanoparticles to stretched DNA strands up to several hundred nanometers in length. As a result, stable and efficient co-expression of GFP and DsRed in porcine kidney PK-15 cells was achieved by magnetofection. The results presented here demonstrate the potential application of magnetic nanoparticles as an attractive delivery system for animal genetics and breeding studies.

  3. Efficient Gene Knockdown in Mouse Oocytes through Peptide Nanoparticle-Mediated SiRNA Transfection

    OpenAIRE

    Jin, Zhen; Li, Ruichao; Zhou, Chunxiang; Shi, Liya; Zhang, Xiaolan; Yang, Zhixia; Zhang, Dong

    2016-01-01

    The use of mouse oocytes as a model for studying female meiosis is very important in reproductive medicine. Gene knockdown by specific small interfering RNA (siRNA) is usually the first step in the study of the function of a target gene in mouse oocytes during in vitro maturation. Traditionally, the only way to introduce siRNA into mouse oocytes is through microinjection, which is certainly less efficient and strenuous than siRNA transfection in somatic cells. Recently, in research using soma...

  4. Suppression of mRNA Nanoparticle Transfection in Human Fibroblasts by Selected Interferon Inhibiting Small Molecule Compounds.

    Science.gov (United States)

    Liu, Yang; Krishnan, Manoj N; Phua, Kyle K L

    2017-07-31

    In vitro transcribed (IVT) mRNA is increasingly applied in lieu of DNA to deliver reprogramming genes to fibroblasts for stem cell derivation. However, IVT mRNA induces interferon (IFN) responses from mammalian cells that reduces transfection efficiency. It has been previously suggested that small molecule inhibitors of IFN are a viable strategy to enhance mRNA transfection efficiency. Herein, we screen a list of commercially available small molecules, including published IFN inhibitors, for their potential to enhance mRNA transfection in BJ fibroblasts. Transfection enhancement is quantified by relative mean fluorescence intensity of translated green fluorescent protein (GFP) in treated cells compared to dimethyl sulfoxide treated controls. Within toxicological constrains, all tested small molecules did not enhance mRNA transfection in BJ fibroblasts while a third of the tested compounds unexpectedly inhibited GFP expression even though IFN-β production is inhibited. Based on the results of our study, we conclude that small molecule inhibitors, including IFN inhibitors, tested in this study do not enhance in vitro mRNA transfection efficiency in human fibroblasts.

  5. Characterization of PEI-coated superparamagnetic iron oxide nanoparticles for transfection: Size distribution, colloidal properties and DNA interaction

    Energy Technology Data Exchange (ETDEWEB)

    Steitz, Benedikt [Laboratory of Powder Technology, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne (Switzerland); Hofmann, Heinrich [Laboratory of Powder Technology, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne (Switzerland); Kamau, Sarah W. [Institute of Veterinary Biochemistry and Molecular Biology, University of Zuerich, Zurich (Switzerland); Hassa, Paul O. [Institute of Veterinary Biochemistry and Molecular Biology, University of Zuerich, Zurich (Switzerland); Hottiger, Michael O. [Institute of Veterinary Biochemistry and Molecular Biology, University of Zuerich, Zurich (Switzerland); Rechenberg, Brigitte von [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Hofmann-Amtenbrink, Magarethe [MatSearch, Chemin Jean Pavillard 14, 1009 Pully (Switzerland); Petri-Fink, Alke [Laboratory of Powder Technology, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne (Switzerland)]. E-mail: alke.fink@epfl.ch

    2007-04-15

    Superparamagnetic iron oxide nanoparticles (SPIONs) were coated with polyethylenimine. Here, we briefly describe the synthesis as well as DNA:PEI:SPION complexes and the characterization of the compounds according to their particle size, {zeta}-potential, morphology, DNA complexing ability, magnetic sedimentation, and colloidal stability. PEI coating of SPIONs led to colloidally stable beads even in high salt concentrations over a wide pH range. DNA plasmids and PCR products encoding for green fluorescent protein were associated with the described beads. The complexes were added to cells and exposed to permanent and pulsating magnetic fields. Presence of these magnetic fields significantly increased the transfection efficiency.

  6. Molecular studies of fibroblasts transfected with hepatitis B virus DNA

    International Nuclear Information System (INIS)

    Chen, M.L.; Hood, A.; Thung, S.N.; Gerber, M.A.

    1987-01-01

    Two subclones (D7 and F8) derived from an NIH 3T3 mouse fibroblast cell line after transfection with hepatitis B virus (HBV) genomes, secreted significantly different amounts of HBsAg and HBeAg. DNA extracted from the subclones revealed only integrated and no extrachromosomal HBV DNA sequences as determined by the Southern blot technique with a /sup 32/P-labeled full length HBV DNA probe. The amount and integration sites of HBV sequences were significantly different in the two subclones. HBV DNA sequences coding for HBsAg and HBcAg were detected by alkaline phosphatase-conjugated, single-stranded synthetic gene-specific oligonucleotide probes revealing a larger number of copies in D7 DNA than in F8 DNA. Using a biotinylated probe for in situ hybridization, HBV DNA was found in the nuclei of all D7 cells with predominant localization to a single chromsome, but only in 10-20% of F8 cells. These observations demonstrate different integration patterns of HBV and DNA in two subclones derived from a transfected cell line and suggest that the amount of integrated HBV DNA is proportional to the amount of HBV antigens produced

  7. Gene delivery using calcium phosphate nanoparticles: Optimization of the transfection process and the effects of citrate and poly(l-lysine) as additives.

    Science.gov (United States)

    Khan, Mohammed A; Wu, Victoria M; Ghosh, Shreya; Uskoković, Vuk

    2016-06-01

    Despite the long history of nanoparticulate calcium phosphate (CaP) as a non-viral transfection agent, there has been limited success in attempts to optimize its properties for transfection comparable in efficiency to that of viral vectors. Here we focus on the optimization of: (a) CaP nanoparticle precipitation conditions, predominantly supersaturation and Ca/P molar ratios; (b) transfection conditions, mainly the concentrations of the carrier and plasmid DNA; (c) the presence of surface additives, including citrate anion and cationic poly(l-lysine) (PLL). CaP nanoparticles significantly improved transfection with plasmid DNA encoding enhanced green fluorescent protein (eGFP) in pre-osteoblastic MC3T3-E1 cells compared to a commercial non-viral carrier. At the same time they elicited significantly lesser cytotoxicity than the commercial carrier. Plasmid DNA acted as a nucleation promoter, decreasing the nucleation lag time of metastable CaP solutions and leading to a higher rate of nucleation and a lower size of the precipitated particles. The degree of supersaturation (DS) of 15 was found to be more optimal for transfection than that of 12.5 or 17.5 and higher. Because CaP particles precipitated at DS 15 were spherical, while DS 17.5 and 21 yielded acicular particles, it was concluded that spherical particle morphologies were more conducive to transfection than the anisotropic ones. Even though the yield at DS 15 was 10 and 100 times lower than that at DS 17.5 and 21, respectively, transfection rates were higher using CaP nanoparticle colloids prepared at DS 15 than using those made at higher or lower DS, indicating that the right particle morphology can outweigh the difference in the amount of the carrier, even when this difference is close to 100×. In contrast to the commercial carrier, the concentration of CaP-pDNA delivered to the cells was directly proportional to the transfection rate. Osteosarcoma K7M2 cells were four times more easily transfectable with

  8. A study on the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles.

    Science.gov (United States)

    Li, Huiling; Chen, Jinwen; Xu, Xuan; Yang, Ruhao; Xiang, Xudong; Zhang, Dongshan

    2016-02-01

    To study the safety and efficiency of the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles, and to evaluate its potential clinical application. The potential and conditions regarding the transfection self-made lipid microbubbles (CY5)-labeled-oligonucleotide (ODN) or CY5-labeled-ODN connective tissue growth factor (CTGF) into the rat kidney were evaluated. Th e safety was evaluated by HE staining, liver and renal function tests. The transfection efficiency was evaluated by fluorescence microscopy. Th e expression of CTGF was detected by RT-PCR and Western blot. Self-made lipid microbubble and/or ultrasound significantly enhanced the efficiency of gene transfer and expression in the kidney. Especially, 85%-90% of total glomerular could be transfected. CY5-labeled-ODN expression could be observed in glomerular, tubular and interstitial area. Th ere was no significant change in blood tests aft er gene transfer. Levels of LDH in 7 days were decreased compared with that at the fi rst day aft er the transfection (Ptransfection of CTGF-antisense-ODN into kidney. The ultrasound-mediated gene transfer by self-made lipid microbubble could enhance the efficiency of ODN and expression in the rat kidney. Th is self-made lipid microbubbles supplement may be use for transfection of target genes.

  9. Inclusion of the helper lipid dioleoyl-phosphatidylethanolamine in solid lipid nanoparticles inhibits their transfection efficiency

    NARCIS (Netherlands)

    de Jesus, Marcelo B.; Radaic, Allan; Hinrichs, Wouter L J; Ferreira, Carmen V; de Paula, Eneida; Hoekstra, Dirk; Zuhorn, Inge S

    Solid lipid nanoparticles (SLNs) are a promising system for the delivery of lipophilic and hydrophilic drugs. They consist of a solid lipid core that is stabilized by a layer of surfactants. By the incorporation of cationic lipids in the formulation, positively charged SLNs can be generated, that

  10. Gene Transfection in High Serum Levels: Case Studies with New Cholesterol Based Cationic Gemini Lipids

    Science.gov (United States)

    Misra, Santosh K.; Biswas, Joydeep; Kondaiah, Paturu; Bhattacharya, Santanu

    2013-01-01

    Background Six new cationic gemini lipids based on cholesterol possessing different positional combinations of hydroxyethyl (-CH2CH2OH) and oligo-oxyethylene -(CH2CH2O)n- moieties were synthesized. For comparison the corresponding monomeric lipid was also prepared. Each new cationic lipid was found to form stable, clear suspensions in aqueous media. Methodology/Principal Findings To understand the nature of the individual lipid aggregates, we have studied the aggregation properties using transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential measurements and X-ray diffraction (XRD). We studied the lipid/DNA complex (lipoplex) formation and the release of the DNA from such lipoplexes using ethidium bromide. These gemini lipids in presence of a helper lipid, 1, 2-dioleoyl phophatidyl ethanol amine (DOPE) showed significant enhancements in the gene transfection compared to several commercially available transfection agents. Cholesterol based gemini having -CH2-CH2-OH groups at the head and one oxyethylene spacer was found to be the most effective lipid, which showed transfection activity even in presence of high serum levels (50%) greater than Effectene, one of the potent commercially available transfecting agents. Most of these geminis protected plasmid DNA remarkably against DNase I in serum, although the degree of stability was found to vary with their structural features. Conclusions/Significance -OH groups present on the cationic headgroups in combination with oxyethylene linkers on cholesterol based geminis, gave an optimized combination of new genera of gemini lipids possessing high transfection efficiency even in presence of very high percentage of serum. This property makes them preferential transfection reagents for possible in vivo studies. PMID:23861884

  11. A comparative study of transfection methods for RNA interference in bone marrow-derived murine dendritic cells

    DEFF Research Database (Denmark)

    Pedersen, Charlotte Demuth; Fang, J J; Pedersen, Anders Elm

    2009-01-01

    Selective gene silencing using RNA interference (RNAi) has been shown to be an efficient method for manipulation of cellular functions. In this study, we compare three previously established methods for transfection of murine bone marrow-derived DC (BM-DC). We tested the efficacy of electroporation...... with the Mouse Nucleofector kit((R)) from Amaxa Biosystems and lipid-based transfection methods using transfection reagents from Santa Cruz Biotechnology or Genlantis. To analyse the transfection efficacy we used FITC-conjugated siRNA as a positive control together with CD80 and CD86 specific siRNA. We show...... that electroporation using the Mouse Nucleofector kit((R)) from Amaxa Biosystems was not an efficient method to transfect BM-DC with siRNA in our hands. Transfection with Santa Cruz Biotechnology reagents resulted in up to 59% FITC-siRNA positive cells, but did not result in effective silencing of CD80 surface...

  12. Cholesterol Domains Enhance Transfection

    Science.gov (United States)

    Betker, Jamie L.; Kullberg, Max; Gomez, Joe; Anchordoquy, Thomas J.

    2014-01-01

    The formation of cholesterol domains in lipoplexes has been associated with enhanced serum stability and transfection rates both in cell culture and in vivo. This study utilizes the ability of saturated phosphatidylcholines to promote the formation of cholesterol domains at much lower cholesterol contents than have been utilized in previous work. The results show that lipoplexes with identical cholesterol and cationic lipid contents exhibit significantly improved transfection efficiencies when a domain is present, consistent with previous work. In addition, studies assessing transfection rates in the absence of serum demonstrate that the ability of domains to enhance transfection is not dependent on interactions with serum proteins. Consistent with this hypothesis, characterization of the adsorbed proteins composing the corona of these lipoplex formulations did not reveal a correlation between transfection and the adsorption of a specific protein. Finally, we show that the interaction with serum proteins can promote domain formation in some formulations, and thereby result in enhanced transfection only after serum exposure. PMID:23557286

  13. Comparative nucleic acid transfection efficacy in primary hepatocytes for gene silencing and functional studies

    Directory of Open Access Journals (Sweden)

    Morral Núria

    2011-01-01

    Full Text Available Abstract Background Primary hepatocytes are the best resource for in vitro studies directed at understanding hepatic processes at the cellular and molecular levels, necessary for novel drug development to treat highly prevalent diseases such as non-alcoholic steatohepatitis, cardiovascular disease and type 2 diabetes. There is a need to identify simple methods to genetically manipulate primary hepatocytes and conduct functional studies with plasmids, small interfering RNA (siRNA or microRNA (miRNA. New lipofection reagents are available that have the potential to yield higher levels of transfection with reduced toxicity. Findings We have tested several liposome-based transfection reagents used in molecular biology research. We show that transfection efficiency with one of the most recently developed formulations, Metafectene Pro, is high with plasmid DNA (>45% cells as well as double stranded RNA (>90% with siRNA or microRNA. In addition, negligible cytotoxicity was present with all of these nucleic acids, even if cells were incubated with the DNA:lipid complex for 16 hours. To provide the proof of concept that these conditions can be used not only for overexpression of a gene of interest, but also in RNA interference applications, we targeted two liver expressed genes, Sterol Regulatory Element-Binding Protein-1 and Fatty Acid Binding Protein 5 using plasmid-mediated short hairpin RNA expression. In addition, similar transfection conditions were used to optimally deliver siRNA and microRNA. Conclusions We have identified a lipid-based reagent for primary hepatocyte transfection of nucleic acids currently used in molecular biology laboratories. The conditions described here can be used to expedite a large variety of research applications, from gene function studies to microRNA target identification.

  14. Antibody transfection into neurons as a tool to study disease pathogenesis.

    Science.gov (United States)

    Douglas, Joshua N; Gardner, Lidia A; Lee, Sangmin; Shin, Yoojin; Groover, Chassidy J; Levin, Michael C

    2012-09-26

    Antibodies provide the ability to gain novel insight into various events taking place in living systems. The ability to produce highly specific antibodies to target proteins has allowed for very precise biological questions to be addressed. Importantly, antibodies have been implicated in the pathogenesis of a number of human diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), paraneoplastic syndromes, multiple sclerosis (MS) and human T-lymphotropic virus type 1 (HTLV-1) associated myelopathy/tropical spastic paraparesis (HAM/TSP). How antibodies cause disease is an area of ongoing investigation, and data suggests that interactions between antibodies and various intracellular molecules results in inflammation, altered cellular messaging, and apoptosis. It has been shown that patients with MS and HAM/TSP produce autoantibodies to the intracellular RNA binding protein heterogeneous ribonuclear protein A1 (hnRNP A1). Recent data indicate that antibodies to both intra-neuronal and surface antigens are pathogenic. Thus, a procedure that allows for the study of intracellular antibody:protein interactions would lend great insight into disease pathogenesis. Genes are commonly transfected into primary cells and cell lines in culture, however transfection of antibodies into cells has been hindered by alteration of antibody structure or poor transfection efficiency. Other methods of transfection include antibody transfection based on cationic liposomes (consisting of DOTAP/DOPE) and polyethylenimines (PEI); both of which resulted in a ten-fold decrease in antibody transfection compared to controls. The method performed in our study is similar to cationic lipid-mediated methods and uses a lipid-based mechanism to form non-covalent complexes with the antibodies through electrostatic and hydrophobic interactions. We utilized Ab-DeliverIN reagent, which is a lipid formulation capable of capturing antibodies through non-covalent electrostatic and

  15. Gene expression profiles in primary duodenal chick cells following transfection with avian influenza virus H5 DNA plasmid encapsulated in silver nanoparticles

    Directory of Open Access Journals (Sweden)

    Jazayeri SD

    2013-02-01

    Full Text Available Seyed Davoud Jazayeri,1 Aini Ideris,1,2 Kamyar Shameli,3 Hassan Moeini,1 Abdul Rahman Omar1,21Institute of Bioscience, 2Faculty of Veterinary Medicine, 3Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, MalaysiaAbstract: In order to develop a systemically administered safe and effective nonviral gene delivery system against avian influenza virus (AIV that induced cytokine expression, the hemagglutinin (H5 gene of AIV, A/Ck/Malaysia/5858/04 (H5N1 and green fluorescent protein were cloned into a coexpression vector pIRES (pIREGFP-H5 and formulated using green synthesis of silver nanoparticles (AgNPs with poly(ethylene glycol and transfected into primary duodenal cells taken from 18-day-old specific-pathogen-free chick embryos. The AgNPs were prepared using moderated temperature and characterized for particle size, surface charge, ultraviolet-visible spectra, DNA loading, and stability. AgNPs and AgNP-pIREGFP-H5 were prepared in the size range of 13.9 nm and 25 nm with a positive charge of +78 ± 0.6 mV and +40 ± 6.2 mV, respectively. AgNPs with a positive surface charge could encapsulate pIREGFP-H5 efficiently. The ultraviolet-visible spectra for AgNP-pIREGFP-H5 treated with DNase I showed that the AgNPs were able to encapsulate pIREGFP-H5 efficiently. Polymerase chain reaction showed that AgNP-pIREGFP-H5 entered into primary duodenal cells rapidly, as early as one hour after transfection. Green fluorescent protein expression was observed after 36 hours, peaked at 48 hours, and remained stable for up to 60 hours. In addition, green fluorescent protein expression generally increased with increasing DNA concentration and time. Cells were transfected using Lipocurax in vitro transfection reagent as a positive control. A multiplex quantitative mRNA gene expression assay in the transfected primary duodenal cells via the transfection reagent and AgNPs with pIREGFP-H5 revealed expression of interleukin (IL-18, IL-15, and IL-12

  16. Biophysical effects in off-resonant gold nanoparticle mediated (GNOME) laser transfection of cell lines, primary- and stem cells using fs laser pulses.

    Science.gov (United States)

    Schomaker, Markus; Killian, Doreen; Willenbrock, Saskia; Heinemann, Dag; Kalies, Stefan; Ngezahayo, Anaclet; Nolte, Ingo; Ripken, Tammo; Junghanß, Christian; Meyer, Heiko; Murua Escobar, Hugo; Heisterkamp, Alexander

    2015-08-01

    Gold nanoparticle mediated (GNOME) laser transfection is a powerful technique to deliver small biologically relevant molecules into cells. However, the transfection of larger and especially negatively charged DNA remains challenging. The efficiency for pDNA was 0.57% using parameter that does not influence the endo- and exogenous DNA. In order to gain a deeper understanding of the actual molecule uptake process, the uptake efficiency was determined using molecules of different sizes. It was evaluated that uncharged dextran molecules (2000 kDa) were delivered with an efficiency of 68%. The intracellular distribution of injected molecules was visualized and larger molecules were primary found in the cytoplasm. Patch clamp measurements suggested a permeabilization time up to 15 minutes. The uptake efficiency depended on the size and charge of the molecule to deliver as well as the cell size. A minor role for transfection plays the cell type since primary stem cells were successfully transfected. The perforation efficiency of semi-adherent and suspension cells is influenced by the cell and molecule size. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Preliminary study of a novel transfection modality for in vivo siRNA delivery to vocal fold fibroblasts.

    Science.gov (United States)

    Kraja, Iv; Bing, Renjie; Hiwatashi, Nao; Rousseau, Bernard; Nalband, Danielle; Kirshenbaum, Kent; Branski, Ryan C

    2017-07-01

    An obstacle to clinical use of RNA-based gene suppression is instability and inefficiency of current delivery modalities. Nanoparticle delivery likely holds great promise, but the kinetics and transfection conditions must be optimized prior to in vivo utility. We investigated a RNA nanoparticle complex incorporating a lipitoid transfection reagent in comparison to a commercially available reagent. In vitro. We investigated which variables influence transfection efficiency of lipitoid oligomers and a commercially available reagent across species, in vitro. These variables included duration, dose, and number of administrations, as well as serum and media conditions. The target gene was Smad3, a signaling protein in the transforming growth factor-β cascade implicated in fibroplasia in the vocal folds and other tissues. The two reagents suppressed Smad3 mRNA for up to 96 hours; lipitoid performed favorably and comparably. Both compounds yielded 60% to 80% mRNA knockdown in rat, rabbit, and human vocal fold fibroblasts (P transfection conditions. These preliminary data are encouraging, and lipitoid warrants further investigation with the goal of clinical utility. NA. Laryngoscope, 127:E231-E237, 2017. © 2016 The American Laryngological, Rhinological and Otological Society, Inc.

  18. Transfection Studies with Colloidal Systems Containing Highly Purified Bipolar Tetraether Lipids from Sulfolobus acidocaldarius

    Science.gov (United States)

    Pinnapireddy, Shashank Reddy; Baghdan, Elias; Jedelská, Jarmila

    2017-01-01

    Lipid vectors are commonly used to facilitate the transfer of nucleic acids into mammalian cells. In this study, two fractions of tetraether lipids from the archaea Sulfolobus acidocaldarius were extracted and purified using different methods. The purified lipid fractions polar lipid fraction E (PLFE) and hydrolysed glycerol-dialkyl-nonitol tetraether (hGDNT) differ in their structures, charge, size, and miscibility from conventional lipids. Liposomes were prepared by mixing tetraether lipids with cholesterol (CH) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) resulting in stable vectors for gene delivery. Lipoplexes were prepared by complexation of liposomes with a luciferase expressing plasmid (pCMV-luc) at certain nitrogen-to-phosphorus (N/P) ratios and optimised for the transient transfection of ovarian adenocarcinoma cells (SK-OV-3). Complexation efficacy was investigated by gel-red fluorescence assay. Biophysical properties, like size, surface charge, and morphology, were investigated by differential light scattering (DLS), atomic force microscopy (AFM), and scanning electron microscopy (Cryo-SEM), respectively, revealing structural differences between liposomes and lipoplexes. A range of stable transfecting agents containing tetraether lipids were obtained by incorporating 5 mol% of tetraether lipids. Lipoplexes showed a decrease in free gel-red with increasing N/P ratios indicating efficient incorporation of plasmid DNA (pDNA) and remarkable stability. Transfection experiments of the lipoplexes revealed successful and superior transfection of SK-OV-3 cell line compared to the commercially available DOTAP and branched polyethyleneimine (25 kDa bPEI). PMID:28239294

  19. Transfection Studies with Colloidal Systems Containing Highly Purified Bipolar Tetraether Lipids from Sulfolobus acidocaldarius

    Directory of Open Access Journals (Sweden)

    Konrad H. Engelhardt

    2017-01-01

    Full Text Available Lipid vectors are commonly used to facilitate the transfer of nucleic acids into mammalian cells. In this study, two fractions of tetraether lipids from the archaea Sulfolobus acidocaldarius were extracted and purified using different methods. The purified lipid fractions polar lipid fraction E (PLFE and hydrolysed glycerol-dialkyl-nonitol tetraether (hGDNT differ in their structures, charge, size, and miscibility from conventional lipids. Liposomes were prepared by mixing tetraether lipids with cholesterol (CH and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP resulting in stable vectors for gene delivery. Lipoplexes were prepared by complexation of liposomes with a luciferase expressing plasmid (pCMV-luc at certain nitrogen-to-phosphorus (N/P ratios and optimised for the transient transfection of ovarian adenocarcinoma cells (SK-OV-3. Complexation efficacy was investigated by gel-red fluorescence assay. Biophysical properties, like size, surface charge, and morphology, were investigated by differential light scattering (DLS, atomic force microscopy (AFM, and scanning electron microscopy (Cryo-SEM, respectively, revealing structural differences between liposomes and lipoplexes. A range of stable transfecting agents containing tetraether lipids were obtained by incorporating 5 mol% of tetraether lipids. Lipoplexes showed a decrease in free gel-red with increasing N/P ratios indicating efficient incorporation of plasmid DNA (pDNA and remarkable stability. Transfection experiments of the lipoplexes revealed successful and superior transfection of SK-OV-3 cell line compared to the commercially available DOTAP and branched polyethyleneimine (25 kDa bPEI.

  20. Ultrasound-targeted transfection of tissue-type plasminogen activator gene carried by albumin nanoparticles to dog myocardium to prevent thrombosis after heart mechanical valve replacement

    Directory of Open Access Journals (Sweden)

    Ji J

    2012-06-01

    Full Text Available Ji Jun, Ji Shang-Yi, Yang Jian-An, He Xia, Yang Xiao-Han, Ling Wen-Ping, Chen Xiao-LingDepartment of Pathology and Cardiovascular Surgery, Shenzhen Sun Yat-Sen Cardiovascular Hospital, Shenzhen, Guangdong, People's Republic of ChinaBackground: There are more than 300,000 prosthetic heart valve replacements each year worldwide. These patients are faced with a higher risk of thromboembolic events after heart valve surgery and long-term or even life-long anticoagulative and antiplatelet therapies are necessary. Some severe complications such as hemorrhaging or rebound thrombosis can occur when the therapy ceases. Tissue-type plasminogen activator (t-PA is a thrombolytic agent. One of the best strategies is gene therapy, which offers a local high expression of t-PA over a prolonged time period to avoid both systemic hemorrhaging and local rebound thrombosis. There are some issues with t-PA that need to be addressed: currently, there is no up-to-date report on how the t-PA gene targets the heart in vivo and the gene vector for t-PA needs to be determined.Aims: To fabricate an albumin nano-t-PA gene ultrasound-targeted agent and investigate its targeting effect on prevention of thrombosis after heart mechanic valve replacement under therapeutic ultrasound.Methods: A dog model of mechanical tricuspid valve replacement was constructed. A highly expressive t-PA gene plasmid was constructed and packaged by nanoparticles prepared with bovine serum albumin. This nanopackaged t-PA gene plasmid was further cross-linked to ultrasonic microbubbles prepared with sucrose and bovine serum albumin to form the ultrasonic-targeted agent for t-PA gene transfection. The agent was given intravenously followed by a therapeutic ultrasound treatment (1 MHz, 1.5 w/cm2, 10 minutes of the heart soon after valve replacement had been performed. The expression of t-PA in myocardium was detected with multiclonal antibodies to t-PA by the indirect immunohistochemical method

  1. [An experimental study on recombinant adenovirus p53 transfected in oral dysplastic epithelial cells].

    Science.gov (United States)

    Xu, Bo; Zhang, Song-Tao; Li, Long-Jiang; Han, Bo; Zhao, Hong-Wei; Pan, Jian

    2009-04-01

    To investigate and evaluate the appropriate virus titer and transfection efficiency of recombinant adenovirus p53 into the oral dysplastic epithelial cells (POE-9n) and provide reference for oral precancerosis research. The transfection sensitivity of adenovirus into oral dysplastic epithelial cells was evaluated by the recombinant adenovirus p53 containing green fluorescent protein (rAd-GFP). Different titre rAd -p53 was transfected into oral dysplastic epithelial cells to evaluate the effects of rAd-p53 on cell proliferation inhibition by MIT assay. The expression of exogenous p53 gene in POE-9n cells was detected by immunocytochemistry. More than 95% POE-9n cells were transfected by rAd-GFP with MOI from 100 to 500 and there was no statistical difference between different MOI values (r=-0.124, P>0.05). It was found that rAd-p53 had significant inhibition effects on POE-9n cell proliferation with MOI from 100 to 500, and there were no significant differences at 96 h and 120 h after the transfection on cell proliferation inhibition (P>0.05). P53 protein was well expressed in rAd-p53 transfected POE-9n cells. Exogenous p53 can be successfully transfected into POE-9n cells by rAd-p53 and the virus titer of MOI 100 was high enough to ensure efficient transfection.

  2. Preliminary study on the mechanism of radioresistance of SHG44 cells transfected by PKB

    International Nuclear Information System (INIS)

    Liu Fenju; Sun Zhiqiang; Yang Xueqin; Jiang Yaqi; Xue Jing

    2007-01-01

    Objective: To explore the radiosensitivity of SHG44 cells increased by suppressing the protein kinase B (PKB), in order to prove whether the PKB activity is related to the radioresistance of SHG44 cells. Methods: PKB gene(pCMV5.HA-m/p-PKBα(PKB), pCMV5.HA-PKBα-DD (T308D/S473D) (PKBD)) were transfected into SHG44 cells by electroporation, the cell proliferation rate was observed among the control, PKB transfected and irradiated groups by MTT assay. The laser confocal microscope was used to detect the changes of cell apoptosis and its microstructure in control, control + radiation, PKB transfected + radiation, PKBD transfected + radiation group. The proliferation of PKB transfected SHG44 cells and the relative factors of inducing apoptosis were analyzed. Results: The plasmid containing extrinsic PKB was successfully transfected into SHG44 cells and expressed PKB mRNA, while there was no expression in the control group; the proliferation rate of transfected SHG44 cells was significantly different from the control group (P 60 Co γ rays could induce SHG44 cell apoptosis with the changes of cell nuclei shape. The SHG44 cells transfected by PKB in the PKB + control group were complete, with few apoptosis cells seen, while the apoptosis was more significant in PKBD + irradiation group comparing to the control-irradiation group. Conclusions: SHG44 cells transfected by PKB could resist the cell apoptosis induced by radiation, suggesting that there were some relations between PKB activity and SHG44 cells radioresistance. (authors)

  3. Improving the osteogenesis of human bone marrow mesenchymal stem cell sheets by microRNA-21-loaded chitosan/hyaluronic acid nanoparticles via reverse transfection

    Directory of Open Access Journals (Sweden)

    Wang Z

    2016-05-01

    Full Text Available Zhongshan Wang,1 Guangsheng Wu,2,3 Mengying Wei,4 Qian Liu,1 Jian Zhou,1 Tian Qin,1 Xiaoke Feng,1 Huan Liu,1 Zhihong Feng,1 Yimin Zhao1 1State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Oral Diseases, Department of Prosthodontics, 2State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Department of Periodontology, School of Stomatology, The Fourth Military Medical University, Xi’an, 3Qingdao First Sanatorium, Jinan Military Region, Qingdao, Shandong Province, 4Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Xi’an, People’s Republic of China Abstract: Cell sheet engineering has emerged as a novel approach to effectively deliver seeding cells for tissue regeneration, and developing human bone marrow mesenchymal stem cell (hBMMSC sheets with high osteogenic ability is a constant requirement from clinics for faster and higher-quality bone formation. In this work, we fabricated biocompatible and safe chitosan (CS/hyaluronic acid (HA nanoparticles (NPs to deliver microRNA-21 (miR-21, which has been proved to accelerate osteogenesis in hBMMSCs; then, the CS/HA/miR-21 NPs were cross-linked onto the surfaces of culture plates with 0.2% gel solution to fabricate miR-21-functionalized culture plates for reverse transfection. hBMMSC sheets were induced continuously for 14 days using a vitamin C-rich method on the miR-21-functionalized culture plates. For the characterization of CS/HA/miR-21 NPs, the particle size, zeta potential, surface morphology, and gel retardation were sequentially investigated. Then, the biological effects of hBMMSC sheets on the miR-21-functionalized culture plates were evaluated. The assay results demonstrated that the hBMMSC sheets could be successfully induced via the novel

  4. Comparative characterization of transfection- and infection-derived simian immunodeficiency virus challenge stocks for in vivo nonhuman primate studies.

    Science.gov (United States)

    Del Prete, Gregory Q; Scarlotta, Matthew; Newman, Laura; Reid, Carolyn; Parodi, Laura M; Roser, James D; Oswald, Kelli; Marx, Preston A; Miller, Christopher J; Desrosiers, Ronald C; Barouch, Dan H; Pal, Ranajit; Piatak, Michael; Chertova, Elena; Giavedoni, Luis D; O'Connor, David H; Lifson, Jeffrey D; Keele, Brandon F

    2013-04-01

    Simian immunodeficiency virus (SIV) stocks for in vivo nonhuman primate models of AIDS are typically generated by transfection of 293T cells with molecularly cloned viral genomes or by expansion in productively infected T cells. Although titers of stocks are determined for infectivity in vitro prior to in vivo inoculation, virus production methods may differentially affect stock features that are not routinely analyzed but may impact in vivo infectivity, mucosal transmissibility, and early infection events. We performed a detailed analysis of nine SIV stocks, comprising five infection-derived SIVmac251 viral swarm stocks and paired infection- and transfected-293T-cell-derived stocks of both SIVmac239 and SIVmac766. Representative stocks were evaluated for (i) virus content, (ii) infectious titer, (iii) sequence diversity and polymorphism frequency by single-genome amplification and 454 pyrosequencing, (iv) virion-associated Env content, and (v) cytokine and chemokine content by 36-plex Luminex analysis. Regardless of production method, all stocks had comparable particle/infectivity ratios, with the transfected-293T stocks possessing the highest overall virus content and infectivity titers despite containing markedly lower levels of virion-associated Env than infection-derived viruses. Transfected-293T stocks also contained fewer and lower levels of cytokines and chemokines than infection-derived stocks, which had elevated levels of multiple analytes, with substantial variability among stocks. Sequencing of the infection-derived SIVmac251 stocks revealed variable levels of viral diversity between stocks, with evidence of stock-specific selection and expansion of unique viral lineages. These analyses suggest that there may be underappreciated features of SIV in vivo challenge stocks with the potential to impact early infection events, which may merit consideration when selecting virus stocks for in vivo studies.

  5. Nanoparticle-Mediated p53 Gene Therapy for Breast Cancer

    National Research Council Canada - National Science Library

    Prabha, Swayam

    2003-01-01

    The effect of different formulation parameters on nanoparticle-mediated gene transfection in vitro was studied Nanoparticles encapsulating plasmid DNA encoding fire fly luciferase were formulated using poly lactide (PLA...

  6. Galectin-1 Reduces Neuroinflammation via Modulation of Nitric Oxide-Arginase Signaling in HIV-1 Transfected Microglia: a Gold Nanoparticle-Galectin-1 "Nanoplex" a Possible Neurotherapeutic?

    Science.gov (United States)

    Aalinkeel, Ravikumar; Mangum, Courtney S; Abou-Jaoude, Eliane; Reynolds, Jessica L; Liu, Maixian; Sundquist, Karin; Parikh, Neil U; Chaves, Lee D; Mammen, Manoj J; Schwartz, Stanley A; Mahajan, Supriya D

    2017-03-01

    Galectins are a family of β-galactoside-binding lectins that are important modulators of homeostasis in the central nervous system (CNS). Galectin-1 is a pivotal regulator of microglia activation that alters the immune balance from neurodegeneration to neuroprotection and could have therapeutic relevance in HIV associated neurocognitive disorders (HAND). We have previously shown that galectin-1 treatment decreased oxidative stress in microglia and hypothesize that the mechanism underlying this phenomenon is the cross regulatory interactions between Nitric oxide (NO) and Arginase I activity in microglia. We induced microglial activation and examined the effect of galectin-1 on the expression of various M1/M2 microglial phenotypic markers. Since, TNF-α is associated with activation of microglial cells involved in pathogenesis of neurodegenerative diseases, we treated HIV transfected human microglial cell cultures (CHME-5/HIV) with TNF-α followed by treatment with galectin-1, to examine the galectin-1 mediated neuro-modulatory response. Our results show that treatment of CHME-5/HIV microglia with galectin-1 reduced TNF-α induced oxidative stress by ~40%, and also significantly reduced iNOS gene expression and NO production while correspondingly increasing arginase-1, cationic amino acid transporter (CAT-1) gene expression and arginase activity. Galectin-1 treatment results in shifting microglia polarization from M1 toward the beneficial M2 phenotype which may prevent neurodegeneration and promote neuroprotection. Thus, our data suggests that galectin-1 treatment reduces neuroinflammation in the CNS microenvironment via the modulation of the NO-arginase network in microglia and thus could play a neuroprotective role in HAND. Further, the therapeutic potential of galectin-1 could be enhanced by conjugation of galectin-1 onto gold nanoparticles (Au-NP), resulting in a nanogold-galectin-1 (Au-Gal-1) multivalent complex that will have more clinical translational

  7. Elucidating the interplay between DNA-condensing and free polycations in gene transfection through a mechanistic study of linear and branched PEI

    DEFF Research Database (Denmark)

    Dai, Zhuojun; Gjetting, Torben; Mattebjerg, Maria Ahlm

    2011-01-01

    In the present study we compare LPEI and BPEI characteristics related to DNA condensation and their role as free polycation chains in gene transfection. Using radioactive 32P labeled DNA, we investigated the effect of free PEI chains on the cellular uptake of polyplexes. Our investigations show...... different properties of BPEI and LPEI polyplexes in condensation and de-condensation processes as well as in cellular uptake, which was tightly correlated with transfection efficiency. In agreement with earlier reports we find all DNA to be condensed at N/P = 3. Further added PEI chains remain free...... in solution. We found that both the cellular uptake and gene transfection of BPEI polyplexes is much more efficient than LPEI polyplexes at a low N/P ratio of 3 (i.e., without free PEI chains). When N/P is high (10, with 7 portions of free PEI), the LPEI and BPEI polyplexes have similar transfection...

  8. Stable transfection of Acanthamoeba.

    Science.gov (United States)

    Yin, J; Henney, H R

    1997-03-01

    The promoter activity of an Acanthamoeba polyubiquitin gene was analyzed in its homologous system. A modified calcium phosphate transfection method using a neomycin marker vector was developed to achieve highly efficient transfection of the Acanthamoeba polyubiquitin gene into Acanthamoeba cells. In this transfection procedure, the calcium phosphate-DNA complex was formed gradually in the medium during incubation with cells and precipitated on the cells. The crucial factors for obtaining efficient transfection were the pH (6.95) of the transfection buffer used for the calcium phosphate precipitation and the amount (25 micrograms/96-well tissue culture plate) and form (circular) of transfecting DNA. Under these conditions, Acanthamoeba isolate 1B6 was transfected at an efficiency of about 40% with the constructed vector pOPSBU, a pOP13CAT-based polyubiquitin gene incorporated neomycin resistance vector. Acanthamoeba polyphaga was transfected at an efficiency of about 10% with this vector. Transfection of both Acanthamoeba strains appeared to result in low copy plasmid integration (about two copies per cell are suggested). The chloramphenicol acetyltransferase (CAT) assays showed that the promoter of the Acanthamoeba polyubiquitin gene in the constructed vector was especially strong in A. polyphaga, thus the pOPSBU-Acanthamoeba system may be useful for the construction of cDNA expression libraries, as well as for the expression of cloned genes.

  9. RNAi in murine hepatocytes: the agony of choice--a study of the influence of lipid-based transfection reagents on hepatocyte metabolism.

    Science.gov (United States)

    Böttger, Jan; Arnold, Katrin; Thiel, Carlo; Rennert, Christiane; Aleithe, Susanne; Hofmann, Ute; Vlaic, Sebastian; Sales, Susanne; Shevchenko, Andrej; Matz-Soja, Madlen

    2015-09-01

    Primary hepatocyte cell cultures are widely used for studying hepatic diseases with alterations in hepatic glucose and lipid metabolism, such as diabetes and non-alcoholic fatty liver disease. Therefore, small interfering RNAs (siRNAs) provide a potent and specific tool to elucidate the signaling pathways and gene functions involved in these pathologies. Although RNA interference (RNAi) in vitro is frequently used in these investigations, the metabolic alterations elucidated by different siRNA delivery strategies have hardly been investigated in transfected hepatocytes. To elucidate the influence of the most commonly used lipid-based transfection reagents on cultured primary hepatocytes, we studied the cytotoxic effects and transfection efficiencies of INTERFERin(®), Lipofectamine(®)RNAiMAX, and HiPerFect(®). All of these transfection agents displayed low cytotoxicity (5.6-9.0 ± 1.3-3.4%), normal cell viability, and high transfection efficiency (fold change 0.08-0.13 ± 0.03-0.05), and they also favored the satisfactory down-regulation of target gene expression. However, when effects on the metabolome and lipidome were studied, considerable differences were observed among the transfection reagents. Cellular triacylglycerides levels were either up- or down-regulated [maximum fold change: INTERFERin(®) (48 h) 2.55 ± 0.34, HiPerFect(®) (24 h) 0.79 ± 0.08, Lipofectamine(®)RNAiMAX (48 h) 1.48 ± 0.21], and mRNA levels of genes associated with lipid metabolism were differentially affected. Likewise, metabolic functions such as amino acid utilization from were perturbed (alanine, arginine, glycine, ornithine, and pyruvate). In conclusion, these findings demonstrate that the choice of non-viral siRNA delivery agent is critical in hepatocytes. This should be remembered, especially if RNA silencing is used for studying hepatic lipid homeostasis and its regulation.

  10. A Model to Study the Phenotypic Changes of Insect Cell Transfection by Copepod Super Green Fluorescent Protein (cop-GFP) in Baculovirus Expression System.

    Science.gov (United States)

    Shokrollahi, Narjes; Shahbazzadeh, Delavar; Pooshang-Bagheri, Kamran; Habibi-Anbouhi, Mahdi; Jahanian-Najafabadi, Ali; Behdani, Mahdi

    2016-07-01

    Baculovirus expression system is one of the most attractive and powerful eukaryotic expression systems for the production of recombinant proteins. The presence of a biomarker is required to monitor transfection efficiency or protein expression levels in insect cells. The aim of this study was to construct a baculovirus expression vector encoding a copepod super green fluorescent protein (copGFP). In this light, the resultant vector was constructed and used for transfection of Spodoptera frugiperda cells. Expression of the copGFP protein in insect cells was confirmed by fluorescent microscopy and Western-blot analysis. The application of copGFP control bacmid can be considered as an appropriate control for insect cell transfection.

  11. Preparation and characterization of polyethylenimine-coated Fe3O4-MCM-48 nanocomposite particles as a novel agent for magnet-assisted transfection.

    Science.gov (United States)

    Yiu, Humphrey H P; McBain, Stuart C; Lethbridge, Zoe A D; Lees, Martin R; Dobson, Jon

    2010-01-01

    A new type of magnetic nanoparticle was synthesized using mesoporous silica MCM-48 as a template. Magnetite (Fe(3)O(4)) nanocrystals were incorporated onto the MCM-48 silica structure by thermal decomposition of iron(III) acetylacetonate. The particle size of these Fe(3)O(4)-MCM-48 composite particles is around 300 nm with an iron oxide content of ca. 20% w/w. Measurements from SQUID magnetometry suggest that these nanoparticles possess superparamagnetic properties similar to those of Fe(3)O(4) nanoparticles. By coating positively charged polyethylenimine on to the surface, DNA can be bound onto the Fe(3)O(4)-MCM-48 nanoparticles. Transfection studies showed that these PEI-Fe(3)O(4)-MCM-48 particles were highly effective as a transfection reagent, and a 400% increase of transfection efficiency compared with the commercial products was recorded.

  12. Graphene based gene transfection

    Science.gov (United States)

    Feng, Liangzhu; Zhang, Shuai; Liu, Zhuang

    2011-03-01

    Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including biomedicine. In this work, for the first time we successfully use graphene as a non-toxic nano-vehicle for efficient gene transfection. Graphene oxide (GO) is bound with cationic polymers, polyethyleneimine (PEI) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI-10k polymer. The positively charged GO-PEI complexes are able to further bind with plasmid DNA (pDNA) for intracellular transfection of the enhanced green fluorescence protein (EGFP) gene in HeLa cells. While EGFP transfection with PEI-1.2k appears to be ineffective, high EGFP expression is observed using the corresponding GO-PEI-1.2k as the transfection agent. On the other hand, GO-PEI-10k shows similar EGFP transfection efficiency but lower toxicity compared with PEI-10k. Our results suggest graphene to be a novel gene delivery nano-vector with low cytotoxicity and high transfection efficiency, promising for future applications in non-viral based gene therapy.Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including biomedicine. In this work, for the first time we successfully use graphene as a non-toxic nano-vehicle for efficient gene transfection. Graphene oxide (GO) is bound with cationic polymers, polyethyleneimine (PEI) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI

  13. Identification of valid reference genes for the normalization of RT-qPCR expression studies in human breast cancer cell lines treated with and without transient transfection.

    Directory of Open Access Journals (Sweden)

    Lin-Lin Liu

    Full Text Available Reverse transcription-quantitative polymerase chain reaction (RT-qPCR is a powerful technique for examining gene expression changes during tumorigenesis. Target gene expression is generally normalized by a stably expressed endogenous reference gene; however, reference gene expression may differ among tissues under various circumstances. Because no valid reference genes have been documented for human breast cancer cell lines containing different cancer subtypes treated with transient transfection, we identified appropriate and reliable reference genes from thirteen candidates in a panel of 10 normal and cancerous human breast cell lines under experimental conditions with/without transfection treatments with two transfection reagents. Reference gene expression stability was calculated using four algorithms (geNorm, NormFinder, BestKeeper and comparative delta Ct, and the recommended comprehensive ranking was provided using geometric means of the ranking values using the RefFinder tool. GeNorm analysis revealed that two reference genes should be sufficient for all cases in this study. A stability analysis suggests that 18S rRNA-ACTB is the best reference gene combination across all cell lines; ACTB-GAPDH is best for basal breast cancer cell lines; and HSPCB-ACTB is best for ER+ breast cancer cells. After transfection, the stability ranking of the reference gene fluctuated, especially with Lipofectamine 2000 transfection reagent in two subtypes of basal and ER+ breast cell lines. Comparisons of relative target gene (HER2 expression revealed different expressional patterns depending on the reference genes used for normalization. We suggest that identifying the most stable and suitable reference genes is critical for studying specific cell lines under certain circumstances.

  14. Comparative study of the transfection efficiency of commonly used viral vectors in rhesus monkey (Macaca mulatta) brains.

    Science.gov (United States)

    Wu, Shi-Hao; Liao, Zhi-Xing; D Rizak, Joshua; Zheng, Na; Zhang, Lin-Heng; Tang, Hen; He, Xiao-Bin; Wu, Yang; He, Xia-Ping; Yang, Mei-Feng; Li, Zheng-Hui; Qin, Dong-Dong; Hu, Xin-Tian

    2017-03-18

    Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non-human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys. The expression of fluorescent cells was used to quantify transfection efficiency. Histological results revealed that recombinant adeno-associated viral vectors, especially the serotype 2/9 coupled with the cytomegalovirus, human synapsin I, or Ca 2+ /calmodulin-dependent protein kinase II promoters, and lentiviral vector coupled with the human ubiquitin C promoter, induced higher expression of fluorescent cells, representing high transfection efficiency. This is the first comparison of transfection efficiencies of different viral vectors carrying different promoters and serotypes in non-human primates (NHPs). These results can be used as an aid to select optimal vectors to transfer exogenous genes into the central nervous system of non-human primates.

  15. SANS Study of Protein Adsorption on Nanoparticles

    Science.gov (United States)

    Kumar, Sugam; Aswal, V. K.; Kohlbrecher, J.

    2011-07-01

    Adsorption of lysozyme protein on silica nanoparticle has been studied using small-angle neutron scattering (SANS) at pH 7. The measurements were carried out on fixed concentration (1 wt %) of nanoparticles and varying concentration of protein in the range 0 to 2 wt%. It has been found that the protein is adsorbed on the nanoparticle surface at very low protein concentrations whereas strong electrostatic interaction of lysozyme with silica nanoparticles at higher protein concentrations leads to the aggregation of nanoparticles. The adsorption is found to be increased with increase in the particle size and the aggregation is determined to be fractal structure.

  16. Enhanced delivery of PEAL nanoparticles with ultrasound targeted microbubble destruction mediated siRNA transfection in human MCF-7/S and MCF-7/ADR cells in vitro

    Directory of Open Access Journals (Sweden)

    Teng Y

    2015-08-01

    Full Text Available Yanwei Teng,1,2,* Min Bai,3,* Ying Sun,2 Qi Wang,1,2 Fan Li,3 Jinfang Xing,3 Lianfang Du,3 Tao Gong,1 Yourong Duan2 1Key Laboratory of Drug Targeting and Novel Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, Sichuan, People’s Republic of China; 2State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 3Department of Ultrasound, Shanghai First People’s Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: The gene knockdown activity of small interfering RNA (siRNA has led to their use as potential therapeutics for a variety of diseases. However, successful gene therapy requires safe and efficient delivery systems. In this study, we choose mPEG-PLGA-PLL nanoparticles (PEAL NPs with ultrasound targeted microbubble destruction (UTMD to efficiently deliver siRNA into cells. An emulsification-solvent evaporation method was used to prepare siRNA-loaded PEAL NPs. The NPs possessed an average size of 132.6±10.3 nm (n=5, with a uniform spherical shape, and had an encapsulation efficiency (EE of more than 98%. As demonstrated by MTT assay, neither PEAL NPs nor siRNA-loaded PEAL NPs showed cytotoxicity even at high concentrations. The results of cellular uptake showed, with the assistance of UTMD, the siRNA-loaded PEAL NPs can be effectively internalized and can subsequently release siRNA in cells. Taken together, PEAL NPs with UTMD may be highly promising for siRNA delivery, making it possible to fully exploit the potential of siRNA-based therapeutics. Keywords: gene delivery, mPEG-PLGA-PLL, UTMD, emulsification-solvent evaporation method, orthogonal design

  17. Optimization of renal transfection using a renal suction-mediated transfection method in mice.

    Science.gov (United States)

    Taniguchi, Yota; Kawakami, Shigeru; Fuchigami, Yuki; Oyama, Natsuko; Yamashita, Fumiyoshi; Konishi, Satoshi; Shimizu, Kazunori; Hashida, Mitsuru

    2016-01-01

    We previously developed a suction-mediated transfection method in mice. The purpose of this study was to optimize the suction-mediated transfection conditions using a pressure-controlled computer system for efficient and safe kidney-targeted gene delivery in mice. Naked pCMV-Luc was injected into the tail vein in mice, and then the right kidney was suctioned by a device of the suction pressure-controlled system. The effects of renal transfection conditions, such as the suction pressure degree, suction pressure waveform and device area were evaluated by measuring luciferase expression. In addition, renal injury was examined. The renal suction-mediated transfection method at -30 kPa showed high transgene expression. The renal suction waveform did not affect the transfection activity. Under the optimized conditions, the high transgene expression was mostly observed at the renal suctioned site. The transfection conditions used did not induce histological defects or increases in two renal injury biomarkers (Kidney injury molecule-1 mRNA and Clusterin mRNA). We have clarified the transfection conditions for efficient and safe transfection in the kidney using the suction-mediated transfection method in mice.

  18. Studies on the biodistribution of dextrin nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Goncalves, C; Gama, F M [IBB-Institute for Biotechnology and Bioengineering, Centre for Biological Engineering, Minho University, Campus de Gualtar, 4710-057 Braga (Portugal); Ferreira, M F M; Martins, J A [Departamento de Quimica, Universidade do Minho, Campus de Gualtar, 4710-057 Braga (Portugal); Santos, A C; Prata, M I M [IBILI, Faculty of Medicine of the University of Coimbra, Coimbra (Portugal); Geraldes, C F G C, E-mail: fmgama@deb.uminho.pt [Departamento de Ciencias da Vida, Faculdade de Ciencia e Tecnologia e Centro de Neurociencias e Biologia Celular, Universidade de Coimbra (Portugal)

    2010-07-23

    The characterization of biodistribution is a central requirement in the development of biomedical applications based on the use of nanoparticles, in particular for controlled drug delivery. The blood circulation time, organ biodistribution and rate of excretion must be well characterized in the process of product development. In this work, the biodistribution of recently developed self-assembled dextrin nanoparticles is addressed. Functionalization of the dextrin nanoparticles with a DOTA-monoamide-type metal chelator, via click chemistry, is described. The metal chelator functionalized nanoparticles were labelled with a {gamma}-emitting {sup 153}Sm{sup 3+} radioisotope and the blood clearance rate and organ biodistribution of the nanoparticles were obtained. The effect of PEG surface coating on the blood clearance rate and organ biodistribution of the nanoparticles was also studied.

  19. Studies on the biodistribution of dextrin nanoparticles

    Science.gov (United States)

    Gonçalves, C.; Ferreira, M. F. M.; Santos, A. C.; Prata, M. I. M.; Geraldes, C. F. G. C.; Martins, J. A.; Gama, F. M.

    2010-07-01

    The characterization of biodistribution is a central requirement in the development of biomedical applications based on the use of nanoparticles, in particular for controlled drug delivery. The blood circulation time, organ biodistribution and rate of excretion must be well characterized in the process of product development. In this work, the biodistribution of recently developed self-assembled dextrin nanoparticles is addressed. Functionalization of the dextrin nanoparticles with a DOTA-monoamide-type metal chelator, via click chemistry, is described. The metal chelator functionalized nanoparticles were labelled with a γ-emitting 153Sm3 + radioisotope and the blood clearance rate and organ biodistribution of the nanoparticles were obtained. The effect of PEG surface coating on the blood clearance rate and organ biodistribution of the nanoparticles was also studied.

  20. Preparation, characterization, and efficient transfection of cationic liposomes and nanomagnetic cationic liposomes

    Directory of Open Access Journals (Sweden)

    Samadikhah HR

    2011-10-01

    Full Text Available Hamid Reza Samadikhah1,*, Asia Majidi2,*, Maryam Nikkhah2, Saman Hosseinkhani11Department of Biochemistry, 2Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran *These authors contributed equally to this work Purpose: Cationic liposomes (CLs are composed of phospholipid bilayers. One of the most important applications of these particles is in drug and gene delivery. However, using CLs to deliver therapeutic nucleic acids and drugs to target organs has some problems, including low transfection efficiency in vivo. The aim of this study was to develop novel CLs containing magnetite to overcome the deficiencies. Patients and methods: CLs and magnetic cationic liposomes (MCLs were prepared using the freeze-dried empty liposome method. Luciferase-harboring vectors (pGL3 were transferred into liposomes and the transfection efficiencies were determined by luciferase assay. Firefly luciferase is one of most popular reporter genes often used to measure the efficiency of gene transfer in vivo and in vitro. Different formulations of liposomes have been used for delivery of different kinds of gene reporters. Lipoplex (liposome–plasmid DNA complexes formation was monitored by gel retardation assay. Size and charge of lipoplexes were determined using particle size analysis. Chinese hamster ovary cells were transfected by lipoplexes (liposome-pGL3; transfection efficiency and gene expression level was evaluated by luciferase assay. Results: High transfection efficiency of plasmid by CLs and novel nanomagnetic CLs was achieved. Moreover, lipoplexes showed less cytotoxicity than polyethyleneimine and Lipofectamine™. Conclusion: Novel liposome compositions (1,2-dipalmitoyl-sn-glycero-3-phosphocholine [DPPC]/dioctadecyldimethylammonium bromide [DOAB] and DPPC/cholesterol/DOAB with high transfection efficiency can be useful in gene delivery in vitro. MCLs can also be used for targeted gene delivery, due to

  1. Enhanced Nanomagnetic Gene Transfection of Human Prenatal Cardiac Progenitor Cells and Adult Cardiomyocytes

    Science.gov (United States)

    Subramanian, Mahendran; Lim, Jenson; Dobson, Jon

    2013-01-01

    Magnetic nanoparticle-based gene transfection has been shown to be an effective, non-viral technique for delivery of both plasmid DNA and siRNA into cells in culture. It has several advantages over other non-viral delivery techniques, such as short transfection times and high cell viability. These advantages have been demonstrated in a number of primary cells and cell lines. Here we report that oscillating magnet array-based nanomagnetic transfection significantly improves transfection efficiency in both human prenatal cardiac progenitor cells and adult cardiomyocytes when compared to static magnetofection, cationic lipid reagents and electroporation, while maintaining high cell viability. In addition, transfection of adult cardiomyocytes was improved further by seeding the cells onto Collagen I-coated plates, with transfection efficiencies of up to 49% compared to 24% with lipid reagents and 19% with electroporation. These results demonstrate that oscillating nanomagnetic transfection far outperforms other non-viral transfection techniques in these important cells. PMID:23936108

  2. Hydrophobic modification of low molecular weight polyethylenimine for improved gene transfection.

    Science.gov (United States)

    Teo, Pei Yun; Yang, Chuan; Hedrick, James L; Engler, Amanda C; Coady, Daniel J; Ghaem-Maghami, Sadaf; George, Andrew J T; Yang, Yi Yan

    2013-10-01

    Hydrophobic modification of low molecular weight (LMW) polyethylenimine (PEI) is known to increase gene transfection efficiency of LMW PEI. However, few studies have explored how the conjugated hydrophobic groups influence the properties of the modified LMW PEI mainly due to difficulties in obtaining well defined final product compositions and limitations in current chemical synthesis routes. The aim of this study was to modify LMW PEI (Mn 1.8 kDa, PEI-1.8) judiciously with different hydrophobic functional groups and to investigate how hydrophobicity, molecular structure and inclusion of hydrogen bonding properties in the conjugated side groups as well as the conjugation degree (number of primary amine groups of PEI-1.8 modified with hydrophobic groups) influence PEI-1.8 gene transfection efficiency. The modified polymers were characterized for DNA binding ability, particle size, zeta potential, in vitro gene transfection efficiency and cytotoxicity in SKOV-3 human ovarian cancer and HepG2 human liver carcinoma cell lines. The study shows that modified PEI-1.8 polymers are able to condense plasmid DNA into cationic nanoparticles, of sizes ~100 nm, whereas unmodified polymer/DNA complexes display larger particle sizes of 2 μm. Hydrophobic modification also increases the zeta potential of polymer/DNA complexes. Importantly, modified PEI-1.8 shows enhanced transfection efficiency over the unmodified counterpart. Higher transfection efficiency is obtained when PEI-1.8 is modified with shorter hydrophobic groups (MTC-ethyl) as opposed to longer ones (MTC-octyl and MTC-deodecyl). An aromatic structured functional group (MTC-benzyl) also enhances transfection efficiency more than an alkyl functional group (MTC-octyl). An added hydrogen-bonding urea group in the conjugated functional group (MTC-urea) does not enhance transfection efficiency over one without urea (MTC-benzyl). The study also demonstrates that modification degree greatly influences gene transfection, and

  3. Surface modification of silica particles with gold nanoparticles as an augmentation of gold nanoparticle mediated laser perforation

    Science.gov (United States)

    Kalies, Stefan; Gentemann, Lara; Schomaker, Markus; Heinemann, Dag; Ripken, Tammo; Meyer, Heiko

    2014-01-01

    Gold nanoparticle mediated (GNOME) laser transfection/perforation fulfills the demands of a reliable transfection technique. It provides efficient delivery and has a negligible impact on cell viability. Furthermore, it reaches high-throughput applicability. However, currently only large gold particles (> 80 nm) allow successful GNOME laser perforation, probably due to insufficient sedimentation of smaller gold nanoparticles. The objective of this study is to determine whether this aspect can be addressed by a modification of silica particles with gold nanoparticles. Throughout the analysis, we show that after the attachment of gold nanoparticles to silica particles, comparable or better efficiencies to GNOME laser perforation are reached. In combination with 1 µm silica particles, we report laser perforation with gold nanoparticles with sizes down to 4 nm. Therefore, our investigations have great importance for the future research in and the fields of laser transfection combined with plasmonics. PMID:25136494

  4. Transfection of Platyhelminthes

    Directory of Open Access Journals (Sweden)

    Bárbara Moguel

    2015-01-01

    Full Text Available Flatworms are one of the most diverse groups within Lophotrochozoa with more than 20,000 known species, distributed worldwide in different ecosystems, from the free-living organisms in the seas and lakes to highly specialized parasites living in a variety of hosts, including humans. Several infections caused by flatworms are considered major neglected diseases affecting countries in the Americas, Asia, and Africa. For several decades, a particular interest on free-living flatworms was due to their ability to regenerate considerable portions of the body, implying the presence of germ cells that could be important for medicine. The relevance of reverse genetics for this group is clear; understanding the phenotypic characteristics of specific genes will shed light on developmental traits of free-living and parasite worms. The genetic manipulation of flatworms will allow learning more about the mechanisms for tissue regeneration, designing new and more effective anthelmintic drugs, and explaining the host-parasite molecular crosstalk so far partially inaccessible for experimentation. In this review, availability of transfection techniques is analyzed across flatworms, from the initial transient achievements to the stable manipulations now developed for free-living and parasite species.

  5. On the role of the colloidal stability of mesoporous silica nanoparticles as gene delivery vectors

    Energy Technology Data Exchange (ETDEWEB)

    Cebrian, Virginia [Hospital Universitario La Paz-IdiPAZ (Spain); Yaguee, Clara; Arruebo, Manuel, E-mail: arruebom@unizar.es [University of Zaragoza, Aragon Nanoscience Institute (INA), C/Mariano Esquillor, Edif. I-D (Spain); Martin-Saavedra, Francisco M. [Hospital Universitario La Paz-IdiPAZ (Spain); Santamaria, Jesus [CIBER de Bioingenieria, Biomateriales y Nanomedicina, CIBER-BBN (Spain); Vilaboa, Nuria [Hospital Universitario La Paz-IdiPAZ (Spain)

    2011-09-15

    Mesoporous silica nanoparticles have been synthesized and functionalized with four different types of molecules containing amino groups, i.e., with primary amines only, with quaternary amines, with quaternized cyclic amines, or with polyethylenimine (PEI), which is formed by primary, secondary, and tertiary amines. These nanoparticles were then incubated with reporter plasmids and the ability of the resulting complexes to transfect human cells was studied. Only nanoparticles functionalized with PEI were efficient for transfection. The agglomeration behavior and the electrokinetic potential of the nanoparticle-plasmid complexes have been studied, as well as their cell internalization behavior using a fluorescent-labeled plasmid that allows its monitorization by confocal microscopy. The results indicate that the efficiency of PEI-functionalized nanoparticles for transfection resides to some extent in the different characteristics imparted to the nanoparticles regarding agglomeration and surface charge behavior.

  6. Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes LipofectamineTM2000.

    Science.gov (United States)

    Huang, Fei; Zhao, Feng; Liang, Li-Ping; Zhou, Mei; Qu, Zhi-Ling; Cao, Yan-Zhen; Lin, Chen

    2015-01-01

    Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Lipofectamine(TM)2000 as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA , miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. The seeding density of Hela cell line and Siha are 1.5 x 10(4) (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (Ptransfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

  7. The Effect of Environmental pH on Polymeric Transfection Efficiency

    OpenAIRE

    Kang, Han Chang; Samsonova, Olga; Kang, Sun-Woong; Bae, You Han

    2011-01-01

    Although polymers, polyplexes, and cells are exposed to various extracellular and intracellular pH environments during polyplex preparation and polymeric transfection, the impact of environmental pH on polymeric transfection has not yet been investigated. This study aims to understand the influence of environmental pH on polymeric transfection by modulating the pH of the transfection medium or the culture medium. Changes in the extracellular pH affected polymeric transfection by way of comple...

  8. Lipid-based Transfection Reagents Exhibit Cryo-induced Increase in Transfection Efficiency

    Directory of Open Access Journals (Sweden)

    Helena Sork

    2016-01-01

    Full Text Available The advantages of lipid-based transfection reagents have permitted their widespread use in molecular biology and gene therapy. This study outlines the effect of cryo-manipulation of a cationic lipid-based formulation, Lipofectamine 2000, which, after being frozen and thawed, showed orders of magnitude higher plasmid delivery efficiency throughout eight different cell lines, without compromising cell viability. Increased transfection efficiency with the freeze-thawed reagent was also seen with 2'-O-methyl phosphorothioate oligonucleotide delivery and in a splice-correction assay. Most importantly, a log-scale improvement in gene delivery using the freeze-thawed reagent was seen in vivo. Using three different methods, we detected considerable differences in the polydispersity of the different nucleic acid complexes as well as observed a clear difference in their surface spreading and sedimentation, with the freeze-thawed ones displaying substantially higher rate of dispersion and deposition on the glass surface. This hitherto overlooked elevated potency of the freeze-thawed reagent facilitates the targeting of hard-to-transfect cells, accomplishes higher transfection rates, and decreases the overall amount of reagent needed for delivery. Additionally, as we also saw a slight increase in plasmid delivery using other freeze-thawed transfection reagents, we postulate that freeze-thawing might prove to be useful for an even wider variety of transfection reagents.

  9. pEGFP transfection into murine skeletal muscle by electrosonoporation

    Science.gov (United States)

    Tamošiūnas, Mindaugas; Jakovels, Dainis; Rubins, Uldis; Kadikis, Roberts; Petrovska, Ramona; Šatkauskas, Saulius

    2017-12-01

    In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can affect the plasmid DNA transfection to mice tibialis cranialis muscle. Multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) fluorescence, providing information on location and duration of EGFP expression. We found that electrosonoporation, commonly enhancing pDNA transfection in vitro, had no positive effect on EGFP transfection efficiency increase in vivo with respect to electroporation alone. We presume that this may be associated with decreased viability of transfected fibers.

  10. Inducement of radionuclides targeting therapy by gene transfection

    International Nuclear Information System (INIS)

    Luo Quanyong

    2001-01-01

    The author presents an overview of gene transfection methods to genetically induce tumor cells to express enhanced levels of cell surface antigens and receptors to intake radiolabeled antibody and peptide targeting and thus increase their therapeutic effect in radiotherapy. The current research include inducement of radioimmunotherapy through CEA gene transfection, inducement of iodine-131 therapy by sodium iodide symporter gene transfection and inducement of MIBG therapy by noradrenaline transporter gene transfection. These studies raise the prospect that gene-therapy techniques could be used to enable the treatment of a wide range of tumors with radiopharmaceuticals of established clinical acceptability

  11. Two skin cell lines from wild-type and albino Japanese flounder (Paralichthys olivaceus): establishment, characterization, virus susceptibility, efficient transfection, and application to albinism study.

    Science.gov (United States)

    Wang, Ruoqing; Zhang, Nianwei; Wang, Renkai; Wang, Shengpeng; Wang, Na

    2017-12-01

    In order to provide an applicable cell platform to study fish pathology and skin pigmentation, two cell lines derived from skin tissues of wild-type and albino Japanese flounder were established and named JFSK_wt and JFSK_alb, respectively. These two cell lines were cultured for 45 passages within approximately 300 days. JFSK_wt and JFSK_alb cells were maintained in Dulbecco's Modified Eagle's Medium and Ham's F-12 Nutrient Mixture (DMEM/F12) supplemented with antibiotics, fetal bovine serum (FBS), 2-mercaptoethanol (2-Me), N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES), and basic fibroblast growth factor (bFGF). The optimal growth temperature for JFSK_wt and JFSK_alb cells was 24 °C, and microscopically, the two cell lines were composed of fibroblast-like cells. Chromosomal analysis revealed that JFSK_wt and JFSK_alb cells had an identical diploid karyotype with 2n = 48t. Results of viral inoculation assays revealed that both cell lines shared similar patterns of viral susceptibility to nervous necrosis virus (NNV). High transfection efficiency was observed in JFSK_wt and JFSK_alb cells transfected with a pEGFP-N3 reporter plasmid and Cy3-siRNA. The detection of dermal marker Dermo-1 showed that these two cells were both derived from the dermis. Finally, three genes involved in the melanogenesis pathway, including adenylate cyclase type 5 (adcy5), microphthalmia-associated transcription factor (mitf), and endothelin B receptor (ednrb), were downregulated in JFSK_alb versus JFSK_wt cells. Thus, the two cell lines, sampled from skin tissue of wild-type and albino Japanese flounder will be not only helpful for fish pathogen research but also beneficial for albinism-related gene function studies.

  12. A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines.

    Science.gov (United States)

    Figueroa, Elizabeth; Bugga, Pallavi; Asthana, Vishwaratn; Chen, Allen L; Stephen Yan, J; Evans, Emily Reiser; Drezek, Rebekah A

    2017-05-02

    Gold-polyamidoamine (AuPAMAM) has previously been shown to successfully transfect cells with high efficiency. However, we have observed that certain cell types are more amenable to Au-PAMAM transfection than others. Here we utilized two representative cell lines-a "difficult to transfect" CT26 cell line and an "easy to transfect" SK-BR3 cell line-and attempted to determine the underlying mechanism for differential transfection in both cell types. Using a commonly established poly-cationic polymer similar to PAMAM (polyethyleneimine, or PEI), we additionally sought to quantify the relative transfection efficiencies of each vector in CT26 and SK-BR3 cells, in the hopes of elucidating any mechanistic differences that may exist between the two transfection vectors. A comparative time course analysis of green fluorescent protein reporter-gene expression and DNA uptake was conducted to quantitatively compare PEI- and AuPAMAM-mediated transfection in CT26 and SK-BR3, while flow cytometry and confocal microscopy were used to determine the contribution of cellular uptake, endosomal escape, and cytoplasmic transport to the overall gene delivery process. Results from the time course analysis and flow cytometry studies revealed that initial complex uptake and cytoplasmic trafficking to the nucleus are likely the two main factors limiting CT26 transfectability. The cell type-dependent uptake and intracellular transport mechanisms impacting gene therapy remain largely unexplored and present a major hurdle in the application-specific design and efficiency of gene delivery vectors. This systematic investigation offers insights into the intracellular mechanistic processes that may account for cell-to-cell differences, as well as vector-to-vector differences, in gene transfectability.

  13. Efficient nanoparticle mediated sustained RNA interference in human primary endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Mukerjee, Anindita; Shankardas, Jwalitha; Ranjan, Amalendu P; Vishwanatha, Jamboor K, E-mail: Jamboor.vishwanatha@unthsc.edu [Department of Molecular Biology and Immunology and Institute for Cancer Research, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Fort Worth, TX 76107 (United States)

    2011-11-04

    Endothelium forms an important target for drug and/or gene therapy since endothelial cells play critical roles in angiogenesis and vascular functions and are associated with various pathophysiological conditions. RNA mediated gene silencing presents a new therapeutic approach to overcome many such diseases, but the major challenge of such an approach is to ensure minimal toxicity and effective transfection efficiency of short hairpin RNA (shRNA) to primary endothelial cells. In the present study, we formulated shAnnexin A2 loaded poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles which produced intracellular small interfering RNA (siRNA) against Annexin A2 and brought about the downregulation of Annexin A2. The per cent encapsulation of the plasmid within the nanoparticle was found to be 57.65%. We compared our nanoparticle based transfections with Lipofectamine mediated transfection, and our studies show that nanoparticle based transfection efficiency is very high ({approx}97%) and is more sustained compared to conventional Lipofectamine mediated transfections in primary retinal microvascular endothelial cells and human cancer cell lines. Our findings also show that the shAnnexin A2 loaded PLGA nanoparticles had minimal toxicity with almost 95% of cells being viable 24 h post-transfection while Lipofectamine based transfections resulted in only 30% viable cells. Therefore, PLGA nanoparticle based transfection may be used for efficient siRNA transfection to human primary endothelial and cancer cells. This may serve as a potential adjuvant treatment option for diseases such as diabetic retinopathy, retinopathy of prematurity and age related macular degeneration besides various cancers.

  14. Efficient nanoparticle mediated sustained RNA interference in human primary endothelial cells

    Science.gov (United States)

    Mukerjee, Anindita; Shankardas, Jwalitha; Ranjan, Amalendu P.; Vishwanatha, Jamboor K.

    2011-11-01

    Endothelium forms an important target for drug and/or gene therapy since endothelial cells play critical roles in angiogenesis and vascular functions and are associated with various pathophysiological conditions. RNA mediated gene silencing presents a new therapeutic approach to overcome many such diseases, but the major challenge of such an approach is to ensure minimal toxicity and effective transfection efficiency of short hairpin RNA (shRNA) to primary endothelial cells. In the present study, we formulated shAnnexin A2 loaded poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles which produced intracellular small interfering RNA (siRNA) against Annexin A2 and brought about the downregulation of Annexin A2. The per cent encapsulation of the plasmid within the nanoparticle was found to be 57.65%. We compared our nanoparticle based transfections with Lipofectamine mediated transfection, and our studies show that nanoparticle based transfection efficiency is very high (~97%) and is more sustained compared to conventional Lipofectamine mediated transfections in primary retinal microvascular endothelial cells and human cancer cell lines. Our findings also show that the shAnnexin A2 loaded PLGA nanoparticles had minimal toxicity with almost 95% of cells being viable 24 h post-transfection while Lipofectamine based transfections resulted in only 30% viable cells. Therefore, PLGA nanoparticle based transfection may be used for efficient siRNA transfection to human primary endothelial and cancer cells. This may serve as a potential adjuvant treatment option for diseases such as diabetic retinopathy, retinopathy of prematurity and age related macular degeneration besides various cancers.

  15. Efficient nanoparticle mediated sustained RNA interference in human primary endothelial cells

    International Nuclear Information System (INIS)

    Mukerjee, Anindita; Shankardas, Jwalitha; Ranjan, Amalendu P; Vishwanatha, Jamboor K

    2011-01-01

    Endothelium forms an important target for drug and/or gene therapy since endothelial cells play critical roles in angiogenesis and vascular functions and are associated with various pathophysiological conditions. RNA mediated gene silencing presents a new therapeutic approach to overcome many such diseases, but the major challenge of such an approach is to ensure minimal toxicity and effective transfection efficiency of short hairpin RNA (shRNA) to primary endothelial cells. In the present study, we formulated shAnnexin A2 loaded poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles which produced intracellular small interfering RNA (siRNA) against Annexin A2 and brought about the downregulation of Annexin A2. The per cent encapsulation of the plasmid within the nanoparticle was found to be 57.65%. We compared our nanoparticle based transfections with Lipofectamine mediated transfection, and our studies show that nanoparticle based transfection efficiency is very high (∼97%) and is more sustained compared to conventional Lipofectamine mediated transfections in primary retinal microvascular endothelial cells and human cancer cell lines. Our findings also show that the shAnnexin A2 loaded PLGA nanoparticles had minimal toxicity with almost 95% of cells being viable 24 h post-transfection while Lipofectamine based transfections resulted in only 30% viable cells. Therefore, PLGA nanoparticle based transfection may be used for efficient siRNA transfection to human primary endothelial and cancer cells. This may serve as a potential adjuvant treatment option for diseases such as diabetic retinopathy, retinopathy of prematurity and age related macular degeneration besides various cancers.

  16. CO oxidation on gold nanoparticles: Theoretical studies

    DEFF Research Database (Denmark)

    Remediakis, Ioannis; Lopez, Nuria; Nørskov, Jens Kehlet

    2005-01-01

    We present a summary of our theoretical results regarding CO oxidation on both oxide-supported and isolated gold nanoparticles. Using Density Functional Theory we have studied the adsorption of molecules and the oxidation reaction of CO on gold clusters. Low-coordinated sites on the gold...... nanoparticles can adsorb small inorganic molecules such as O2 and CO, and the presence of these sites is the key factor for the catalytic properties of supported gold nanoclusters. Other contributions, induced by the presence of the support, can provide parallel channels for the reaction and modulate the final...

  17. Improved transfection of HUVEC and MEF cells using DNA ...

    Indian Academy of Sciences (India)

    Cells such as mouse embryonic fibroblasts (MEFs) and human umbilical vein endothelial cells (HUVECs) used in stem cell research and endothelial cell physiology and pathology studies are difficult to transfect using 'standard' nonviral transfection methods. We have developed a novel gene delivery technique, which uses ...

  18. On the role of the colloidal stability of mesoporous silica nanoparticles as gene delivery vectors

    International Nuclear Information System (INIS)

    Cebrián, Virginia; Yagüe, Clara; Arruebo, Manuel; Martín-Saavedra, Francisco M.; Santamaría, Jesus; Vilaboa, Nuria

    2011-01-01

    Mesoporous silica nanoparticles have been synthesized and functionalized with four different types of molecules containing amino groups, i.e., with primary amines only, with quaternary amines, with quaternized cyclic amines, or with polyethylenimine (PEI), which is formed by primary, secondary, and tertiary amines. These nanoparticles were then incubated with reporter plasmids and the ability of the resulting complexes to transfect human cells was studied. Only nanoparticles functionalized with PEI were efficient for transfection. The agglomeration behavior and the electrokinetic potential of the nanoparticle–plasmid complexes have been studied, as well as their cell internalization behavior using a fluorescent-labeled plasmid that allows its monitorization by confocal microscopy. The results indicate that the efficiency of PEI-functionalized nanoparticles for transfection resides to some extent in the different characteristics imparted to the nanoparticles regarding agglomeration and surface charge behavior.

  19. Evaluation of the magnetic field requirements for nanomagnetic gene transfection

    Science.gov (United States)

    Fouriki, A.; Farrow, N.; Clements, M.A.; Dobson, J.

    2010-01-01

    The objective of this work was to examine the effects of magnet distance (and by proxy, field strength) on nanomagnetic transfection efficiency. Methods non-viral magnetic nanoparticle-based transfection was evaluated using both static and oscillating magnet arrays. Results Fluorescence intensity (firefly luciferase) of transfected H292 cells showed no increase using a 96-well NdFeB magnet array when the magnets were 5 mm from the cell culture plate or nearer. At 6 mm and higher, fluorescence intensity decreased systematically. Conclusion In all cases, fluorescence intensity was higher when using an oscillating array compared to a static array. For distances closer than 5 mm, the oscillating system also outperformed Lipofectamine 2000™. PMID:22110859

  20. Evaluation of the magnetic field requirements for nanomagnetic gene transfection

    Directory of Open Access Journals (Sweden)

    A. Fouriki

    2010-07-01

    Full Text Available The objective of this work was to examine the effects of magnet distance (and by proxy, field strength on nanomagnetic transfection efficiency. Methods: non-viral magnetic nanoparticle-based transfection was evaluated using both static and oscillating magnet arrays. Results: Fluorescence intensity (firefly luciferase of transfected H292 cells showed no increase using a 96-well NdFeB magnet array when the magnets were 5 mm from the cell culture plate or nearer. At 6 mm and higher, fluorescence intensity decreased systematically. Conclusion: In all cases, fluorescence intensity was higher when using an oscillating array compared to a static array. For distances closer than 5 mm, the oscillating system also outperformed Lipofectamine 2000™.

  1. From oleic acid-capped iron oxide nanoparticles to polyethyleneimine-coated single-particle magnetofectins

    Energy Technology Data Exchange (ETDEWEB)

    Cruz-Acuña, Melissa [University of Florida, J. Crayton Pruitt Family Department of Biomedical Engineering (United States); Maldonado-Camargo, Lorena [University of Florida, Department of Chemical Engineering (United States); Dobson, Jon; Rinaldi, Carlos, E-mail: carlos.rinaldi@bme.ufl.edu [University of Florida, J. Crayton Pruitt Family Department of Biomedical Engineering (United States)

    2016-09-15

    Various inorganic nanoparticle designs have been developed and used as non-viral gene carriers. Magnetic gene carriers containing polyethyleneimine (PEI), a well-known transfection agent, have been shown to improve DNA transfection speed and efficiency in the presence of applied magnetic field gradients that promote particle–cell interactions. Here we report a method to prepare iron oxide nanoparticles conjugated with PEI that: preserves the narrow size distribution of the nanoparticles, conserves magnetic properties throughout the process, and results in efficient transfection. We demonstrate the ability of the particles to electrostatically bind with DNA and transfect human cervical cancer (HeLa) cells by the use of an oscillating magnet array. Their transfection efficiency is similar to that of Lipofectamine 2000™, a commercial transfection reagent. PEI-coated particles were subjected to acidification, and acidification in the presence of salts, before DNA binding. Results show that although these pre-treatments did not affect the ability of particles to bind DNA they did significantly enhanced transfection efficiency. Finally, we show that these magnetofectins (PEI-MNP/DNA) complexes have no effect on the viability of cells at the concentrations used in the study. The systematic preparation of magnetic vectors with uniform physical and magnetic properties is critical to progressing this non-viral transfection technology.

  2. From oleic acid-capped iron oxide nanoparticles to polyethyleneimine-coated single-particle magnetofectins

    International Nuclear Information System (INIS)

    Cruz-Acuña, Melissa; Maldonado-Camargo, Lorena; Dobson, Jon; Rinaldi, Carlos

    2016-01-01

    Various inorganic nanoparticle designs have been developed and used as non-viral gene carriers. Magnetic gene carriers containing polyethyleneimine (PEI), a well-known transfection agent, have been shown to improve DNA transfection speed and efficiency in the presence of applied magnetic field gradients that promote particle–cell interactions. Here we report a method to prepare iron oxide nanoparticles conjugated with PEI that: preserves the narrow size distribution of the nanoparticles, conserves magnetic properties throughout the process, and results in efficient transfection. We demonstrate the ability of the particles to electrostatically bind with DNA and transfect human cervical cancer (HeLa) cells by the use of an oscillating magnet array. Their transfection efficiency is similar to that of Lipofectamine 2000™, a commercial transfection reagent. PEI-coated particles were subjected to acidification, and acidification in the presence of salts, before DNA binding. Results show that although these pre-treatments did not affect the ability of particles to bind DNA they did significantly enhanced transfection efficiency. Finally, we show that these magnetofectins (PEI-MNP/DNA) complexes have no effect on the viability of cells at the concentrations used in the study. The systematic preparation of magnetic vectors with uniform physical and magnetic properties is critical to progressing this non-viral transfection technology.

  3. Transfection of isolated rainbow trout, Oncorhynchus mykiss, granulosa cells through chemical transfection and electroporation at 12°C.

    Science.gov (United States)

    Marivin, E; Mourot, B; Loyer, P; Rime, H; Bobe, J; Fostier, A

    2015-09-15

    Over-expression or inhibition of gene expression can be efficiently used to analyse the functions and/or regulation of target genes. Modulation of gene expression can be achieved through transfection of exogenous nucleic acids into target cells. Such techniques require the development of specific protocols to transfect cell cultures with nucleic acids. The aim of this study was to develop a method of transfection suitable for rainbow trout granulosa cells in primary culture. After the isolation of rainbow trout granulosa cells, chemical transfection of cells with a fluorescent morpholino oligonucleotide (MO) was tested using FuGENE HD at 12 °C. Electroporation was also employed to transfect these cells with either a plasmid or MO. Transfection was more efficient using electroporation (with the following settings: 1200 V/40 ms/1p) than chemical transfection, but electroporation by itself was deleterious, resulting in a decrease of the steroidogenic capacity of the cells, measured via estradiol production from its androgenic substrate. The disturbance of cell biology induced by the transfection method per se should be taken into account in data interpretation when investigating the effects of under- or over-expression of candidate genes. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Highly efficient transfection of human THP-1 macrophages by nucleofection.

    Science.gov (United States)

    Maeß, Marten B; Wittig, Berith; Lorkowski, Stefan

    2014-09-02

    Macrophages, as key players of the innate immune response, are at the focus of research dealing with tissue homeostasis or various pathologies. Transfection with siRNA and plasmid DNA is an efficient tool for studying their function, but transfection of macrophages is not a trivial matter. Although many different approaches for transfection of eukaryotic cells are available, only few allow reliable and efficient transfection of macrophages, but reduced cell vitality and severely altered cell behavior like diminished capability for differentiation or polarization are frequently observed. Therefore a transfection protocol is required that is capable of transferring siRNA and plasmid DNA into macrophages without causing serious side-effects thus allowing the investigation of the effect of the siRNA or plasmid in the context of normal cell behavior. The protocol presented here provides a method for reliably and efficiently transfecting human THP-1 macrophages and monocytes with high cell vitality, high transfection efficiency, and minimal effects on cell behavior. This approach is based on Nucleofection and the protocol has been optimized to maintain maximum capability for cell activation after transfection. The protocol is adequate for adherent cells after detachment as well as cells in suspension, and can be used for small to medium sample numbers. Thus, the method presented is useful for investigating gene regulatory effects during macrophage differentiation and polarization. Apart from presenting results characterizing macrophages transfected according to this protocol in comparison to an alternative chemical method, the impact of cell culture medium selection after transfection on cell behavior is also discussed. The presented data indicate the importance of validating the selection for different experimental settings.

  5. Acoustic Liquid Handling for Rapid siRNA Transfection Optimization.

    Science.gov (United States)

    Xiao, Andrew S; Lightcap, Eric S; Bouck, David C

    2015-09-01

    Gene knockdown by small interfering RNA (siRNA) has been used extensively to investigate the function of genes in targeted and genome-wide studies. One of the primary challenges of siRNA studies of any scale is to achieve sufficient gene knockdown to produce the biological changes that lead to measurable phenotypes. Reverse, lipid-based transfection efficiency minimally requires the optimization of the following parameters: cell number, knockdown duration, siRNA oligonucleotide concentration, type/brand of transfection lipid, and transfection lipid concentration. In this study, we describe a methodology to utilize the flexibility and low-volume range of the Echo acoustic liquid handler to rapidly screen a matrix of transfection conditions. The matrix includes six different transfection lipids from three separate vendors across a broad range of concentrations. Our results validate acoustic liquid transfer for the delivery of siRNAs and transfection reagents. Finally, this methodology is applied to rapidly optimize transfection conditions across many tissue culture cell lines derived from various originating tissues. © 2015 Society for Laboratory Automation and Screening.

  6. AGONIST ANTAGONIST INTERACTIONS WITH CLONED HUMAN 5-HT1A RECEPTORS - VARIATIONS IN INTRINSIC ACTIVITY STUDIED IN TRANSFECTED HELA-CELLS

    NARCIS (Netherlands)

    BODDEKE, HWGM; FARGIN, A; RAYMOND, J.; SCHOEFFTER, P; HOYER, D

    The characteristics of 5-HT1A-recognition sites and receptor-mediated release of intracellular calcium were established in two transfected HeLa cell lines (HA 6 and HA 7) expressing different levels of human 5-HT1A receptors (about 3000 and 500 fmol/mg protein, Fargin et al. 1989; 1991; Raymond et

  7. Agonist/antagonist interactions with cloned human 5-HT(1A) receptors: Variations in intrinsic activity studied in transfected HeLa cells

    NARCIS (Netherlands)

    Boddeke, H.W.G.M.; Fargin, A.; Raymond, J.R.; Schoeffter, P.; Hoyer, D.

    1992-01-01

    The characteristics of 5-HT(1A)-recognition sites and receptor-mediated release of intracellular calcium were established in two transfected HeLa cell lines (HA 6 and HA 7) expressing different levels of human 5-HT(1A) receptors (about 3000 and 500 fmol/mg protein, Fargin et al. 1989; 1991; Raymond

  8. Enhancement of DNA-transfection frequency by X-rays

    International Nuclear Information System (INIS)

    Iwamoto, Ryota; Fushimi, Kazuo; Hiraki, Yoshio; Namba, Masayoshi

    1997-01-01

    This study was conducted to evaluate the frequency of DNA transfection into human cells following X-ray irradiation. We transfected plasmid DNA (pSV2neo) into human cells, HeLa and PA-1, by either calcium phosphate precipitation or the lipofection method immediately after irradiating the cells with various doses of X-rays. The transfection frequency was evaluated by counting the number of G418-resistant colonies. When circular plasmid DNA was used, irradiation up to a dose of 2 Gy dose-dependently increased the transfection frequency, which reached a maximum of 5 to 10-fold that of the control unirradiated cells. When linear plasmid DNA was used, the transfection frequency was 2 times higher than that of circular DNA. All five of the clones that were randomly chosen expressed the transfected neo gene. In addition, the pSV2neo gene was randomly integrated into the genomic DNA of each clone. These findings indicate that X-ray treatment can facilitate foreign DNA transfer into human cells and that radiation-induced DNA breaks may promote the insertion of foreign DNA into host DNA. The enhancement of DNA transfection with X-rays may be instrumental in practicing gene therapy. (author)

  9. Optical sorting and photo-transfection of mammalian cells

    CSIR Research Space (South Africa)

    Mthunzi, P

    2010-02-01

    Full Text Available ), embryonic kidney, Chinese hamster ovary as well as pluripotent stem cells using a tightly focused titanium sapphire femtosecond pulsed laser beam spot. These investigations permitted advanced biological studies in femtosecond laser transfection: firstly...

  10. Comparison of different cationized proteins as biomaterials for nanoparticle-based ocular gene delivery.

    Science.gov (United States)

    Zorzi, Giovanni K; Párraga, Jenny E; Seijo, Begoña; Sanchez, Alejandro

    2015-11-01

    Cationized polymers have been proposed as transfection agents for gene therapy. The present work aims to improve the understanding of the potential use of different cationized proteins (atelocollagen, albumin and gelatin) as nanoparticle components and to investigate the possibility of modulating the physicochemical properties of the resulting nanoparticle carriers by selecting specific protein characteristics in an attempt to improve current ocular gene-delivery approaches. The toxicity profiles, as well as internalization and transfection efficiency, of the developed nanoparticles can be modulated by modifying the molecular weight of the selected protein and the amine used for cationization. The most promising systems are nanoparticles based on intermediate molecular weight gelatin cationized with the endogenous amine spermine, which exhibit an adequate toxicological profile, as well as effective association and protection of pDNA or siRNA molecules, thereby resulting in higher transfection efficiency and gene silencing than the other studied formulations. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Physical properties of Cu nanoparticles: A molecular dynamics study

    Energy Technology Data Exchange (ETDEWEB)

    Kart, H.H., E-mail: hkart@pau.edu.tr [Department of Physics, Pamukkale University, Kınıklı Campus, 20017 Denizli (Turkey); Yildirim, H.; Ozdemir Kart, S. [Department of Physics, Pamukkale University, Kınıklı Campus, 20017 Denizli (Turkey); Çağin, T. [Department of Materials Science and Engineering, Texas A and M University, College Station, TX 77845-3003 (United States); Department of Chemical Engineering, Texas A and M University, College Station, TX 77845-3122 (United States)

    2014-09-15

    Thermodynamical, structural and dynamical properties of Cu nanoparticles are investigated by using Molecular Dynamics (MD) simulations at various temperatures. In this work, MD simulations of the Cu-nanoparticles are performed by means of the MPiSiM codes by utilizing from Quantum Sutton-Chen (Q-SC) many-body force potential to define the interactions between the Cu atoms. The diameters of the copper nanoparticles are varied from 2 nm to 10 nm. MD simulations of Cu nanoparticles are carried out at low and high temperatures to study solid and liquid properties of Cu nanoparticles. Simulation results such as melting point, radial distribution function are compared with the available experimental bulk results. Radial distribution function, mean square displacement, diffusion coefficient, Lindemann index and Honeycutt–Andersen index are also calculated for estimating the melting point of the Copper nanoparticles. - Highlights: • Solid and liquid properties of Cu nanoparticles are studied. • Molecular dynamics utilizing the Quantum Sutton Chen potential is used in this work. • Melting temperatures of nanoparticles are strongly depended on nanoparticle sizes. • Heat capacity, radial distribution function and diffusion coefficients are studied. • Structures of nanoparticles are analyzed by Lindemann and Honeycutt–Andersen index.

  12. Physical properties of Cu nanoparticles: A molecular dynamics study

    International Nuclear Information System (INIS)

    Kart, H.H.; Yildirim, H.; Ozdemir Kart, S.; Çağin, T.

    2014-01-01

    Thermodynamical, structural and dynamical properties of Cu nanoparticles are investigated by using Molecular Dynamics (MD) simulations at various temperatures. In this work, MD simulations of the Cu-nanoparticles are performed by means of the MPiSiM codes by utilizing from Quantum Sutton-Chen (Q-SC) many-body force potential to define the interactions between the Cu atoms. The diameters of the copper nanoparticles are varied from 2 nm to 10 nm. MD simulations of Cu nanoparticles are carried out at low and high temperatures to study solid and liquid properties of Cu nanoparticles. Simulation results such as melting point, radial distribution function are compared with the available experimental bulk results. Radial distribution function, mean square displacement, diffusion coefficient, Lindemann index and Honeycutt–Andersen index are also calculated for estimating the melting point of the Copper nanoparticles. - Highlights: • Solid and liquid properties of Cu nanoparticles are studied. • Molecular dynamics utilizing the Quantum Sutton Chen potential is used in this work. • Melting temperatures of nanoparticles are strongly depended on nanoparticle sizes. • Heat capacity, radial distribution function and diffusion coefficients are studied. • Structures of nanoparticles are analyzed by Lindemann and Honeycutt–Andersen index

  13. Hydrophobically modified chitosan/gold nanoparticles for DNA delivery

    International Nuclear Information System (INIS)

    Bhattarai, Shanta Raj; Remant Bahadur, K.C.; Aryal, Santosh; Bhattarai, Narayan; Kim, Sun Young; Yi, Ho Keun; Hwang, Pyoung Han; Kim, Hak Yong

    2008-01-01

    Present study dealt an application of modified chitosan gold nanoparticles (Nac-6-Au) for the immobilization of necked plasmid DNA. Gold nanoparticles stabilized with N-acylated chitosan were prepared by graft-onto approach. The stabilized gold nanoparticles were characterized by different physico-chemical techniques such as UV-vis, TEM, ELS and DLS. MTT assay was used for in vitro cytotoxicity of the nanoparticles into three different cell lines (NIH 3T3, CT-26 and MCF-7). The formulation of plasmid DNA with the nanoparticles corresponds to the complex forming capacity and in-vitro/in-vivo transfection efficiency was studied via gel electrophoresis and transfection methods, respectively. Results showed the modified chitosan gold nanoparticles were well-dispersed and spherical in shape with average size around 10∼12 nm in triple distilled water at pH 7.4, and showed relatively no cytotoxicity at low concentration. Addition of plasmid DNA on the aqueous solution of the nanoparticles markedly reduced surface potential (50.0∼66.6%) as well as resulted in a 13.33% increase in hydrodynamic diameters of the formulated nanoparticles. Transfection efficiency of Nac-6-Au/DNA was dependent on cell type, and higher β-galactosidase activity was observed on MCF-7 breast cancer cell. Typically, this activity was 5 times higher in 4.5 mg/ml nanoparticles concentration than that achieved by the nanoparticles of other concentrations (and/or control). However, this activity was lower in in-vitro and dramatically higher in in-vivo than that of commercially available transfection kit (Lipofectin (registered) ) and DNA. From these results, it can be expected to develop alternative new vectors for gene delivery

  14. Hydrophobically modified chitosan/gold nanoparticles for DNA delivery

    Energy Technology Data Exchange (ETDEWEB)

    Bhattarai, Shanta Raj; Remant Bahadur, K.C.; Aryal, Santosh [Chonbuk National University, Department of Bionanosystem Engineering (Korea, Republic of); Bhattarai, Narayan [University of Washington, Department of Materials Science and Engineering (United States); Kim, Sun Young [Chonbuk National University, Department of Pediatrics, School of Medicine (Korea, Republic of); Yi, Ho Keun [Chonbuk National University, Department of Biochemistry, School of Dentisty (Korea, Republic of); Hwang, Pyoung Han [Chonbuk National University, Department of Pediatrics, School of Medicine (Korea, Republic of); Kim, Hak Yong [Chonbuk National University, Department of Textile Engineering (Korea, Republic of)], E-mail: khy@moak.chonbuk.ac.kr

    2008-01-15

    Present study dealt an application of modified chitosan gold nanoparticles (Nac-6-Au) for the immobilization of necked plasmid DNA. Gold nanoparticles stabilized with N-acylated chitosan were prepared by graft-onto approach. The stabilized gold nanoparticles were characterized by different physico-chemical techniques such as UV-vis, TEM, ELS and DLS. MTT assay was used for in vitro cytotoxicity of the nanoparticles into three different cell lines (NIH 3T3, CT-26 and MCF-7). The formulation of plasmid DNA with the nanoparticles corresponds to the complex forming capacity and in-vitro/in-vivo transfection efficiency was studied via gel electrophoresis and transfection methods, respectively. Results showed the modified chitosan gold nanoparticles were well-dispersed and spherical in shape with average size around 10{approx}12 nm in triple distilled water at pH 7.4, and showed relatively no cytotoxicity at low concentration. Addition of plasmid DNA on the aqueous solution of the nanoparticles markedly reduced surface potential (50.0{approx}66.6%) as well as resulted in a 13.33% increase in hydrodynamic diameters of the formulated nanoparticles. Transfection efficiency of Nac-6-Au/DNA was dependent on cell type, and higher {beta}-galactosidase activity was observed on MCF-7 breast cancer cell. Typically, this activity was 5 times higher in 4.5 mg/ml nanoparticles concentration than that achieved by the nanoparticles of other concentrations (and/or control). However, this activity was lower in in-vitro and dramatically higher in in-vivo than that of commercially available transfection kit (Lipofectin (registered) ) and DNA. From these results, it can be expected to develop alternative new vectors for gene delivery.

  15. Synergistic effects of ultrasound-targeted microbubble destruction and TAT peptide on gene transfection: an experimental study in vitro and in vivo.

    Science.gov (United States)

    Zhou, Zhiyi; Zhang, Ping; Ren, Jianli; Ran, Haitao; Zheng, Yuanyi; Li, Pan; Zhang, Qunxia; Zhang, Maohui; Wang, Zhigang

    2013-09-28

    Cell-permeable peptides (CPPs) and ultrasound-targeted microbubble destruction (UTMD) have tremendous potential for gene delivery. However, their applications are limited due to nonspecificity of CPPs and low transfection efficiency of UTMD. Here, we developed a 'smart' gene delivery system by encapsulating TAT peptide (TATp) and hepatocyte growth factor (HGF) gene within lipid microbubbles, in which TATp was protected from being enzymatically cleaved and HGF gene was protected from degradation. This new strategy had synergistic effects of UTMD and TATp on gene transfection. We investigated the efficacy and safety of HGF gene transfection mediated by the combination of UTMD and TATp in vitro and in vivo. The results from MTT assay and flow cytometry analyses indicated that the combination of UTMD and TATp could enhance HGF gene expression in HUVECs without any significant side effect on cell viability. In rat myocardial infarction models, we demonstrated that the protein and mRNA expressions of HGF in myocardium caused by the combination of UTMD and TATp were the highest. Histopathological findings demonstrated that the combination of UTMD and TATp enhanced myocardial microvasculature and ameliorated myocardial fibrosis. In conclusion, the combination of UTMD and TATp might be a safe and efficient technique for gene delivery. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Ligand-Modified Human Serum Albumin Nanoparticles for Enhanced Gene Delivery.

    Science.gov (United States)

    Look, Jennifer; Wilhelm, Nadine; von Briesen, Hagen; Noske, Nadja; Günther, Christine; Langer, Klaus; Gorjup, Erwin

    2015-09-08

    The development of nonviral gene delivery systems is a great challenge to enable safe gene therapy. In this study, ligand-modified nanoparticles based on human serum albumin (HSA) were developed and optimized for an efficient gene therapy. Different glutaraldehyde cross-linking degrees were investigated to optimize the HSA nanoparticles for gene delivery. The peptide sequence arginine-glycine-aspartate (RGD) and the HIV-1 transactivator of transduction sequence (Tat) are well-known as promising targeting ligands. Plasmid DNA loaded HSA nanoparticles were covalently modified on their surface with these different ligands. The transfection potential of the obtained plasmid DNA loaded RGD- and Tat-modified nanoparticles was investigated in vitro, and optimal incubation conditions for these preparations were studied. It turned out that Tat-modified HSA nanoparticles with the lowest cross-linking degree of 20% showed the highest transfection potential. Taken together, ligand-functionalized HSA nanoparticles represent promising tools for efficient and safe gene therapy.

  17. Ac magnetic susceptibility study of in vivo nanoparticle biodistribution

    Energy Technology Data Exchange (ETDEWEB)

    Gutierrez, L; Veintemillas-Verdaguer, S; Serna, C J; Morales, M P [Instituto de Ciencia de Materiales de Madrid, ICMM-CSIC, Sor Juana Ines de la Cruz 3, Cantoblanco 28049, Madrid (Spain); MejIas, R; Barber, D F [Centro Nacional de BiotecnologIa, CNB-CSIC, Darwin 3, Cantoblanco 28049, Madrid (Spain); Lazaro, F J, E-mail: lucia@icmm.csic.es [Departamento de Ciencia y Tecnologia de Materiales y Fluidos, Universidad de Zaragoza, Maria de Luna 3, 50018, Zaragoza (Spain)

    2011-06-29

    We analysed magnetic nanoparticle biodistribution, before and after cytokine conjugation, in a mouse model by ac susceptibility measurements of the corresponding resected tissues. Mice received repeated intravenous injections of nanoparticle suspension for two weeks and they were euthanized 1 h after the last injection. In general, only 10% of the total injected nanoparticles after multiple exposures were found in tissues. The rest of the particles may probably be metabolized or excreted by the organism. Our findings indicate that the adsorption of interferon to DMSA-coated magnetic nanoparticles changes their biodistribution, reducing the presence of nanoparticles in lungs and therefore their possible toxicity. The specific targeting of the particles to tumour tissues by the use of an external magnetic field has also been studied. Magnetic nanoparticles were observed by transmission electron microscopy in the targeted tissue and quantified by ac magnetic susceptibility.

  18. Neurotensin-Conjugated Reduced Graphene Oxide with Multi-Stage Near-Infrared-Triggered Synergic Targeted Neuron Gene Transfection In Vitro and In Vivo for Neurodegenerative Disease Therapy.

    Science.gov (United States)

    Hsieh, Tsung-Ying; Huang, Wei-Chen; Kang, Yi-Da; Chu, Chao-Yi; Liao, Wen-Lin; Chen, You-Yin; Chen, San-Yuan

    2016-12-01

    Delivery efficiency with gene transfection is a pivotal point in achieving maximized therapeutic efficacy and has been an important challenge with central nervous system (CNS) diseases. In this study, neurotensin (NT, a neuro-specific peptide)-conjugated polyethylenimine (PEI)-modified reduced graphene oxide (rGO) nanoparticles with precisely controlled two-stage near-infrared (NIR)-laser photothermal treatment to enhance the ability to target neurons and achieve high gene transfection in neurons. First-stage NIR laser irradiation on the cells with nanoparticles attached on the surface can increase the permeability of the cell membrane, resulting in an apparent increase in cellular uptake compared to untreated cells. In addition, second-stage NIR laser irradiation on the cells with nanoparticles inside can further induce endo/lysosomal cavitation, which not only helps nanoparticles escape from endo/lysosomes but also prevents plasmid DNA (pDNA) from being digested by DNase I. At least double pDNA amount can be released from rGO-PEI-NT/pDNA under NIR laser trigger release compared to natural release. Moreover, in vitro differentiated PC-12 cell and in vivo mice (C57BL/6) brain transfection experiments have demonstrated the highest transfection efficiency occurring when NT modification is combined with external multi-stage stimuli-responsive NIR laser treatment. The combination of neuro-specific targeting peptide and external NIR-laser-triggered aid provides a nanoplatform for gene therapy in CNS diseases. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. The Effect of Environmental pH on Polymeric Transfection Efficiency

    Science.gov (United States)

    Kang, Han Chang; Samsonova, Olga; Kang, Sun-Woong; Bae, You Han

    2011-01-01

    Although polymers, polyplexes, and cells are exposed to various extracellular and intracellular pH environments during polyplex preparation and polymeric transfection, the impact of environmental pH on polymeric transfection has not yet been investigated. This study aims to understand the influence of environmental pH on polymeric transfection by modulating the pH of the transfection medium or the culture medium. Changes in the extracellular pH affected polymeric transfection by way of complex factors such as pH-induced changes in polymer characteristics (e.g., proton buffering capacity and ionization), polyplex characteristics (e.g., size, surface charge, and decomplexation), and cellular characteristics (e.g., cellular uptake, cell cycle phases, and intracellular pH environment). Notably, acidic medium delayed endocytosis, endosomal acidification, cytosolic release, and decomplexation of polyplexes, thereby negatively affecting gene expression. However, acidic medium inhibited mitosis and reduced dilution of gene expression, resulting in increased transfection efficiency. Compared to pH 7.4 medium, acidic transfection medium reduced gene expression 1.6~7.7-fold whereas acidic culture medium enhanced transfection efficiency 2.1~2.6-fold. Polymeric transfection was affected more by the culture medium than by the transfection medium. Understanding the effects of extracellular pH during polymeric transfection may stimulate new strategies for determining effective and safe polymeric gene carriers. PMID:22130563

  20. QCM-D study of nanoparticle interactions.

    Science.gov (United States)

    Chen, Qian; Xu, Shengming; Liu, Qingxia; Masliyah, Jacob; Xu, Zhenghe

    2016-07-01

    Quartz crystal microbalance with dissipation monitoring (QCM-D) has been proven to be a powerful research tool to investigate in situ interactions between nanoparticles and different functionalized surfaces in liquids. QCM-D can also be used to quantitatively determine adsorption kinetics of polymers, DNA and proteins from solutions on various substrate surfaces while providing insights into conformations of adsorbed molecules. This review aims to provide a comprehensive overview on various important applications of QCM-D, focusing on deposition of nanoparticles and attachment-detachment of nanoparticles on model membranes in complex fluid systems. We will first describe the working principle of QCM-D and DLVO theory pertinent to understanding nanoparticle deposition phenomena. The interactions between different nanoparticles and functionalized surfaces for different application areas are then critically reviewed. Finally, the potential applications of QCM-D in other important fields are proposed and knowledge gaps are identified. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Fast and Efficient Transfection of Mouse Embryonic Stem Cells Using Non-Viral Reagents.

    Science.gov (United States)

    Tamm, Christoffer; Kadekar, Sandeep; Pijuan-Galitó, Sara; Annerén, Cecilia

    2016-10-01

    Reliable and efficient DNA and RNA transfection methods are required when studying the role of individual genes in mouse pluripotent stem cells. However, these cells usually grow in tight clusters and are therefore more difficult to transfect than many other cell lines. We have found that transfection is especially challenging when mouse embryonic stem (mES) cells are cultured in the newly described 2i medium, which is based on two chemical inhibitors of differentiation pathways. In the present study we have performed a side-by-side comparison of commercially available, non-viral transfection reagents with regard to their ability to deliver plasmid DNA and siRNA into adherent and/or trypsinized mES cells cultured in 2i medium, assessing transfection rates, plasmid gene expression, siRNA mediated knockdown of Oct4 and viability. Finally, we present a fast and efficient method for transfection of trypsinized mES cells using the liposomal-based Lipofectamine 2000. With only a five-minute long transfection time we obtained at least 85 % transfected cells with 80 % maintained viability. Moreover, this protocol saves up to a day of experimental time since the cells are in suspension at the time of transfection, which allows for immediately re-plating into the appropriate format. This fast, simplified and highly efficient transfection method will be valuable for both basic research and high-throughput applications.

  2. Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering

    Science.gov (United States)

    2017-01-01

    The efficient delivery of foreign nucleic acids (transfection) into cells is a critical tool for fundamental biomedical research and a pillar of several biotechnology industries. There are currently three main strategies for transfection including reagent, instrument, and viral based methods. Each technology has significantly advanced cell transfection; however, reagent based methods have captured the majority of the transfection market due to their relatively low cost and ease of use. This general method relies on the efficient packaging of a reagent with nucleic acids to form a stable complex that is subsequently associated and delivered to cells via nonspecific electrostatic targeting. Reagent transfection methods generally use various polyamine cationic type molecules to condense with negatively charged nucleic acids into a highly positively charged complex, which is subsequently delivered to negatively charged cells in culture for association, internalization, release, and expression. Although this appears to be a straightforward procedure, there are several major issues including toxicity, low efficiency, sorting of viable transfected from nontransfected cells, and limited scope of transfectable cell types. Herein, we report a new strategy (SnapFect) for nucleic acid transfection to cells that does not rely on electrostatic interactions but instead uses an integrated approach combining bio-orthogonal liposome fusion, click chemistry, and cell surface engineering. We show that a target cell population is rapidly and efficiently engineered to present a bio-orthogonal functional group on its cell surface through nanoparticle liposome delivery and fusion. A complementary bio-orthogonal nucleic acid complex is then formed and delivered to which chemoselective click chemistry induced transfection occurs to the primed cell. This new strategy requires minimal time, steps, and reagents and leads to superior transfection results for a broad range of cell types

  3. Interactions of nanomaterials with biological systems: A study of bio-mineralized nanoparticles and nanoparticle antibiotics

    Science.gov (United States)

    Gifford, Jennifer Chappell

    Nature is continually able to out-perform laboratory syntheses of nanomaterials with control of specific properties under ambient temperatures, pressures and pH. The investigation of existing biomolecule-mediated nanoparticle synthesis provides insight and knowledge necessary for duplicating these processes. In this way, peptides or proteins with nanomaterial mediation capabilities can be: 1) explored to further understand the ways in which biomolecules create specific nanoparticles then 2) used to create genetically encodable tags for use in electron tomography. The goal of designing such a tag was to assist in closing the resolution gap that exists in current imaging techniques between approximately 5 nm and 100 nm. Presented in this thesis are examples of peptides and proteins that form iron oxide, silver or gold nanoparticles under discrete circumstances. Three iron oxide-related bacterial proteins -- bacterioferritin, Dps and Mms6 -- were investigated for potential use. Similarly, a silver mineralizing peptide, Ge8, was studied upon attachment to the filamentous protein, FtsZ, and a gold mineralizing peptide, A3, was examined to characterize the way in which it mediates the formation of both Au0 nanoclusters and nanoparticles. Given the established interactions that occur between nanoparticles and biomolecules, it may not be surprising that gold nanoparticles displaying specific ratios of functional groups are able to interact with bacteria, in some cases inhibiting growth or causing cell death as antibiotics. A previously developed small molecule variable ligand display (SMVLD) method was expanded to identify a nanoparticle conjugate with a minimal inhibitory concentration (MIC99.9) of 6 muM for Mycobacterium smegmatis, a common laboratory model for M. tuberculosis and the first example of SMVLD applied to mycobacteria. Nanoparticle structure-activity relationships, modes of action and approximations of mammalian cell toxicities were also explored to expand

  4. [Optimization of triple plasmids transfection into HEK293 cells mediated by polyethylenimine].

    Science.gov (United States)

    Fu, Qiang; Li, Yan; Zheng, Zhaofen; Liu, Aizhong; Yuan, Zhenhua; Peng, Jianqiang; He, Jin

    2015-02-01

    In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.

  5. Paclitaxel tumor priming promotes delivery and transfection of intravenous lipid-siRNA in pancreatic tumors.

    Science.gov (United States)

    Wang, Jie; Lu, Ze; Wang, Junfeng; Cui, Minjian; Yeung, Bertrand Z; Cole, David J; Wientjes, M Guillaume; Au, Jessie L-S

    2015-10-28

    The major barrier for using small interfering RNA (siRNA) as cancer therapeutics is the inadequate delivery and transfection in solid tumors. We have previously shown that paclitaxel tumor priming, by inducing apoptosis, expands the tumor interstitial space, improves the penetration and dispersion of nanoparticles and siRNA-lipoplexes in 3-dimensional tumor histocultures, and promotes the delivery and transfection efficiency of siRNA-lipoplexes under the locoregional setting in vivo (i.e., intraperitoneal treatment of intraperitoneal tumors). The current study evaluated whether tumor priming is functional for systemically delivered siRNA via intravenous injection, which would subject siRNA to several additional delivery barriers and elimination processes. We used the same pegylated cationic (PCat)-siRNA lipoplexes as in the intraperitoneal study to treat mice bearing subcutaneous human pancreatic Hs766T xenograft tumors. The target gene was survivin, an inducible chemoresistance gene. The results show single agent paclitaxel delayed tumor growth but also significantly induced the survivin protein level in residual tumors, whereas addition of PCat-siSurvivin completely reversed the paclitaxel-induced survivin and enhanced the paclitaxel activity (ppriming, by promoting the interstitial transport and cytoplasmic release, is critical to promote the delivery and transfection of siRNA in vivo. In addition, because paclitaxel has broad spectrum activity and is used to treat multiple types of solid tumors including the hard-to-treat pancreatic cancer, the synergistic paclitaxel+siSurvivin combination represents a potentially useful chemo-gene therapy. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Repair of ionizing radiation damage in primate αDNA transfected into rat cells

    International Nuclear Information System (INIS)

    Bases, R.; Mendez, F.

    1992-01-01

    The time-course of repair of irradiated primate αDNA was studied after transfection and recovery from rat NRK cells. Rat cells were chosen for transfection because they have no αDNA. Plasmid pBUC4α10, containing 10 tandem 172 bp αDNA subunits in its 5kbp DNA, was irradiated and introduced into the rat cells by electroporation. The transfected αDNA was then recovered from NRK nuclei free of extraneous rat DNA, permitting study of the fate of the transfected αDNA in time-course experiments. αDNA continuously entered nuclei for processing in the first 2.5h after transfection. The pool of damaged bases in αDNA in NRK nuclei was detectable 2.5 h after transfection. (author)

  7. Establishment of transient and stable transfection systems for Babesia ovata.

    Science.gov (United States)

    Hakimi, Hassan; Yamagishi, Junya; Kegawa, Yuto; Kaneko, Osamu; Kawazu, Shin-Ichiro; Asada, Masahito

    2016-03-23

    Bovine babesiosis is a tick-borne disease caused by several species of Babesia which produce acute and fatal disease in cattle and affect livestock industry worldwide. Babesia ovata is a benign species widespread in east Asian countries and causes anemia, particularly in cattle which are co-infected with Theileria orientalis. The development of genetic manipulation methods is necessary to improve our understanding of the basic biology of protozoan pathogens toward a better control of disease. Such tools have not been developed for B. ovata, and are the aim of this study. In this study we transfected constructs that were designed to evaluate the ability of several B. ovata promoter candidates to drive expression of a reporter luciferase. We found that the elongation factor-1 alpha intergenic region (ef-1α IG) and the actin 5' non-coding region (NR) had highest promoter activities. To establish a stable transfection system, we generated a plasmid construct in which the ef-1α IG promoter drives gfp expression, and the actin 5' NR mediates expression of the selectable marker hdhfr. The plasmid was designed for episomal transfection, as well as to integrate by double cross-over homologous recombination into the ef-1α locus. Circular or linearized plasmid was transfected by electroporation into in vitro cultured B. ovata and retention of the plasmid was facilitated by drug selection with 5 nM WR99210 initiated 48 h after transfection. After one-week cultivation with WR99210, GFP-expressing parasites were observed by fluorescence microscopy. Integration of the plasmid construct into the ef-1α locus was confirmed by PCR, Southern blot analysis, and sequencing of recombination sites. These results confirm successful development of a stable transfection system for B. ovata. The current study provides a fundamental molecular tool to aid in molecular and cellular studies of B. ovata.

  8. Theoretical Studies of Optical Properties of Silver Nanoparticles

    International Nuclear Information System (INIS)

    Ye-Wan, Ma; Zhao-Wang, Wu; Li-Hua, Zhang; Jie, Zhang

    2010-01-01

    Optical properties of silver nanoparticles such as extinction, absorption and scattering efficiencies are studied based on Green's function theory. The numerical simulation results show that optical properties of silver nanoparticles are mainly dependent on their sizes and geometries; the localized plasmon resonance peak is red shifted when the dielectric constant of the particle's surrounding medium increases or when a substrate is presented. The influences of wave polarizations, the incident angles of light, the composite silver and multiply-layers on the plasmon resonance are also reported. The numerical simulation of optical spectra is a very useful tool for nanoparticle growth and characterization. (fundamental areas of phenomenology(including applications))

  9. Aging study of the powdered magnetite nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Khan, Umar Saeed, E-mail: omar_aps@yahoo.co.uk [Department of Physics, University of Peshawar (Pakistan); Rahim, Abdur, E-mail: rahimkhan533@gmail.com [Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology, Lahore (Pakistan); Khan, Nasrullah [Department of Physics, Kohat University of Science and Technology, Kohat (Pakistan); Muhammad, Nawshad; Rehman, Fozia [Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology, Lahore (Pakistan); Ahmad, Khalid [Institute of Chemistry, State University of Campinas, PO Box 6154, 13083-970 Campinas, SP (Brazil); Iqbal, Jibran [College of Natural and Health Sciences, Zayed University, 144534 Abu Dhabi (United Arab Emirates)

    2017-03-01

    Magnetite nanoparticles were produced via co-precipitation method and then stored at room temperature for 6 years in aerobic atmosphere. Variations in the inherent solid phase and solid interfacial properties of the prepared magnetite nanoparticles were investigated. For this purpose the fresh and aged samples were characterized using transmission electron microscopy, vibrating sample magnetometer, X-ray diffractometer and energy dispersive X-ray spectrometer. The solid phase transformations of magnetite nanoparticles to maghemite nanoparticles as well as formation of other iron oxides were happened. After aging of 6 years, no change was occurred in the magnetic features; however increase in particle size from 9.6 to 18.5 measured by transmission electron microscopy was confirmed. The crystallite size and vibrating sample magnetometer values were measured before and after aging and found to increase from 8.98 nm and 47.23 emu/g to 16.18 nm and 58.36 emu/g respectively. The formation of other iron oxides, recrystallization and agglomeration during aging process, caused a significant decrease in the specific surface area from 124.43 to 45.00 m{sup 2}/g of the stored sample. - Highlights: • Magnetite nanoparticles (NPs) were produced via co-precipitation method. • Inherent solid phase and interfacial properties of NP were evaluated after 6 years. • The solid phase transformations of magnetite NPs to maghemite NPs was happened. • After aging of 6 years, no change was occurred in the magnetic features.

  10. Construction of spherical liposome on solid transducers for electrochemical DNA sensing and transfection.

    Science.gov (United States)

    Bhuvana, Mohanlal; Dharuman, Venkataraman

    2014-10-01

    Cationic 1,2-dioleoyl trimethyl ammonium propane (DOTAP) and neutral 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) are anchored on cysteamine (cyst), mercaptopropionic acid (MPA) monolayer (thiol monolayers) modified on an individual gold transducer. DOTAP and DOPE are mixed with gold nanoparticle (AuNP) to form spherical liposome-AuNP. The electrochemical behaviors of the surface attached DOTAP-AuNP and DOPE-AuNP in presence of [Fe(CN)6](3-/4-) depend on the method of layer formation. Transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), and ultraviolet (UV)-visible spectroscopic techniques are used to characterize the liposome-AuNP nanocomposite. The studies indicate stability of spherical liposome-AuNP on the gold transducer. Label-free DNA hybridization detection on these surfaces reveals different detection limits. Confocal laser scanning microscopy (CLSM) is used to confirm the cell transfection.

  11. Size dependent studies of metal nanoparticles with bio-fluorophores

    Science.gov (United States)

    Patil, Ajeetkumar; Ballary, Steffy; George, Sajan D.; Chidangil, Santhosh

    2017-06-01

    Interaction of noble metal nanoparticles (NPs) with fluorophores has been an important research area in the field of material science and biomedical field. In the proximity of a metal nanoparticle, there is a quenching or enhancement in the intrinsic fluorescence of the fluorophore . The conditional quenching of the fluorescence can be used for negative sensing whereas enhancement in the fluorescence can be used to gain greater sensitivity and high signal to noise ratio in the molecular sensing/imaging. The current work deals with the systematic studies to understand the fluorescence quenching for few bio-fluorophores (NADH and FAD) when interacted with different sized silver nano-particles of (10nm, 40nm and 100nm). Home assembled Laser Induced Fluorescence (LIF) set-up was used to study the fluorescence quenching of NADH and FAD for different sized silver nanoparticles.

  12. A Systematic Structure-Activity Study of a New Type of Small Peptidic Transfection Vector Reveals the Importance of a Special Oxo-Anion-Binding Motif for Gene Delivery.

    Science.gov (United States)

    Junghänel, Sandra; Karczewski, Sarah; Bäcker, Sandra; Knauer, Shirley K; Schmuck, Carsten

    2017-11-16

    We discovered a new class of artificial peptidic transfection vectors based on an artificial anion-binding motif, the guanidiniocarbonylpyrrole (GCP) cation. This new type of vector is surprisingly smaller than traditional systems, and our previous work suggested that the GCP group was important for promoting critical endosomal escape. We now present here a systematic comparison of similar DNA ligands featuring our GCP oxo-anion-binding motif with DNA ligands only consisting of naturally occurring amino acids. Structure-activity studies showed that the artificial binding motif clearly outperformed natural amino acids such as histidine, lysine, and arginine. It improved the ability to shuttle foreign genetic material into cells, yet successfully mediated endosomal escape. Also, plasmids that were complexed by our artificial ligands were stabilized against cytosolic degradation to some extent. This resulted in the successful expression of plasmid information (comparable to gold standards such as polyethyleneimine). Hence, our study clearly demonstrates the importance of the tailor-made GCP anion-binding site for efficient gene transfection. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Nonlinear optical studies of single gold nanoparticles

    NARCIS (Netherlands)

    Dijk, Meindert Alexander van

    2007-01-01

    Gold nanoparticles are spherical clusters of gold atoms, with diameters typically between 1 and 100 nanometers. The applications of these particles are rather diverse, from optical labels for biological experiments to data carrier for optical data storage. The goal of my project was to develop new

  14. Polymeric nanoparticles stabilized by surfactants: kinetic studies

    Czech Academy of Sciences Publication Activity Database

    Pánek, Jiří; Filippov, Sergey K.; Koňák, Čestmír; Steinhart, Miloš; Štěpánek, Petr

    2011-01-01

    Roč. 32, č. 8 (2011), s. 1105-1110 ISSN 0193-2691 R&D Projects: GA ČR GAP208/10/1600 Institutional research plan: CEZ:AV0Z40500505 Keywords : nanoparticles * solvent shifting * time-resolved SAXS Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 0.560, year: 2011

  15. High-Efficient Transfection of Human Embryonic Stem Cells by Single-Cell Plating and Starvation.

    Science.gov (United States)

    Liu, Hui; Ren, Caiping; Zhu, Bin; Wang, Lei; Liu, Weidong; Shi, Jia; Lin, Jianxing; Xia, Xiaomeng; Zeng, Fei; Chen, Jiawen; Jiang, Xingjun

    2016-03-15

    Nowadays, the low efficiency of small interfering RNA (siRNA) or plasmid DNA (pDNA) transfection is a critical issue in genetic manipulation of human embryonic stem (hES) cells. Development of an efficient transfection method for delivery of siRNAs and plasmids into hES cells becomes more and more imperative. In this study, we tried to modify the traditional transfection protocol by introducing two crucial processes, single-cell plating and starvation, to increase the transfection efficiency in hES cells. Furthermore, we comparatively examined the transfection efficiency of some commercially available siRNA or pDNA transfection reagents in hES cells. Our results showed that the new developed method markedly enhanced the transfection efficiency without influencing the proliferation and pluripotency of hES cells. Lipofectamine RNAiMAX exhibited much higher siRNA transfection efficiency than the other reagents, and FuGENE HD was identified as the best suitable reagent for efficient pDNA transfection of hES cells among the tested reagents.

  16. Optimization of in vitro culture and transfection condition of bovine ...

    African Journals Online (AJOL)

    The present study aimed to optimize the in vitro culture and transfection efficiency of bovine primary spermatogonial stem cells (SSCs). To this end, SSCs were obtained from newborn Holstein bull calves by two-step enzymatic digestion. After enrichment and culture, SSCs were characterized by using alkaline phosphatase ...

  17. A robust transfection reagent for the transfection of CHO and HEK293 cells and production of recombinant proteins and lentiviral particles - PTG1.

    Science.gov (United States)

    Gonçalves, Cristine; Gross, Fabian; Guégan, Philippe; Cheradame, Hervé; Midou, Patrick

    2014-11-01

    Bioproduction of recombinant proteins (r-proteins) and recombinant lentiviral particles (r-lentiviral particles) requires robust transfections consisting of efficient protocols that are easy to implement, with good reproducibility for a maximum production of proteins and lentiviral particles in a short time with low cytotoxicity. This study evaluates the capacity of histidinylated polyethyleneimine I (PTG1) to facilitate robust DNA transfection, with low cytotoxicity, of Chinese hamster ovary (CHO) and human embryonic kidney (HEK293T) cells for the production of r-proteins and r-lentiviral particles. We report that PTG1 transfection of cells in suspension with a plasmid DNA encoding enhanced green fluorescent protein leads to 72 and 97% of transfected CHO and HEK293T cells respectively, and does not significantly affect cell viability. PTG1 transfection of 100 mL of CHO-S cell culture in suspension at a cell density of 2 × 10(6) cells /mL resulted in a high level of transfected cells and protein expression after transfection with 0.75 μg/mL plasmid DNA. Transfection with PTG1 is more efficient than LipofectAmine2000™, and gene expression is higher than observed with FreeStyle™ and JetPEI®. Tri-transfection of HEK293T packaging cells leads to the production of a higher level of r-lentiviral particles compared to the calcium phosphate method, and permits two harvests of viral particles within three days. These results show that PTG1 is a powerful new transfection reagent for cell lines frequently used for recombinant protein and lentiviral particle production. PTG1 could be used in protocols for bioproduction of therapeutic proteins such as antibodies for cancer treatments and viral vectors for gene therapy applications. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. In silico study of nanoparticles in soft matter environments

    Science.gov (United States)

    Ranatunga, R. J. K. Udayana

    Nanoparticles have unique, size dependent properties which arise due to their small physical dimensions. These unique properties imbue nanoparticles with outstanding potential in most fields of modern technology. However, successful application of nanoparticles is predicated on controlling their synthesis, self-assembly and environmental impact. Achieving this control requires an understanding of both the chemical reactivity and physical behavior of nanoparticles. This is particularly true for soft matter applications, where the dynamic and deformable nature of the environment complicates characterization. This dissertation aims to investigate physical properties of several nanoparticle-soft matter systems using molecular dynamics computer simulations. By utilizing simulations, spatial and temporal resolutions unavailable to microscopy are accessed, enabling us to observe molecular behavior and calculate thermodynamic properties through statistical mechanics. We begin by studying nanoparticles grafted with soft surfactant ligands, known to spontaneously localize at fluid interfaces. We show that conventional theories treating the energetics of particle localization fail at the nanoscale when the particle shape is deformable. Morever, we show that free surfactants and nanoparticles exhibit synergy in lowering the oil-water interfacial tension, and propose a simple mechanism for this behavior. Computer models allow us to consider different nanoparticle geometries by using simple continuum solids. This treatment yields analytical interaction potentials useful in probing nanoparticle behavior in their native environments. Spherical fullerenes can be approximated with a hollow shell, allowing the investigation of fullerene effects on lipid bilayers as a function of particle size. Carbon nanotubes can also be approximated with a cylinder allowing us to study the stability of nanotube dispersions in aqueous media. Simulations can also be utilized to investigate the role

  19. Transfection of Sertoli cells with androgen receptor alters gene expression without androgen stimulation.

    Science.gov (United States)

    Fietz, D; Markmann, M; Lang, D; Konrad, L; Geyer, J; Kliesch, S; Chakraborty, T; Hossain, H; Bergmann, M

    2015-12-29

    Androgens play an important role for the development of male fertility and gained interest as growth and survival factors for certain types of cancer. Androgens act via the androgen receptor (AR/Ar), which is involved in various cell biological processes such as sex differentiation. To study the functional mechanisms of androgen action, cell culture systems and AR-transfected cell lines are needed. Transfection of AR into cell lines and subsequent gene expression analysis after androgen treatment is well established to investigate the molecular biology of target cells. However, it remains unclear how the transfection with AR itself can modulate the gene expression even without androgen stimulation. Therefore, we transfected Ar-deficient rat Sertoli cells 93RS2 by electroporation using a full length human AR. Transfection success was confirmed by Western Blotting, immunofluorescence and RT-PCR. AR transfection-related gene expression alterations were detected with microarray-based genome-wide expression profiling of transfected and non-transfected 93RS2 cells without androgen stimulation. Microarray analysis revealed 672 differentially regulated genes with 200 up- and 472 down-regulated genes. These genes could be assigned to four major biological categories (development, hormone response, immune response and metabolism). Microarray results were confirmed by quantitative RT-PCR analysis for 22 candidate genes. We conclude from our data, that the transfection of Ar-deficient Sertoli cells with AR has a measurable effect on gene expression even without androgen stimulation and cause Sertoli cell damage. Studies using AR-transfected cells, subsequently stimulated, should consider alterations in AR-dependent gene expression as off-target effects of the AR transfection itself.

  20. D-Glucosamine Promotes Transfection Efficiency during Electroporation

    Directory of Open Access Journals (Sweden)

    Kazunari Igawa

    2014-01-01

    Full Text Available D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually used to effectively introduce foreign genes into tissue culture cells. Buffers for electroporation with or without D-glucosamine are used in experiments of transfection vectors. This is the first study to indirectly observe the stability and protection of the osteoblast membrane against both electric stress and gene uptake (the proton sponge hypothesis: osmotic rupture during endosomes prior to fusion with lysosomes in electroporation with D-glucosamine application. The transfection efficiency was evaluated as the fluorescence intensity of the transfected green fluorescent protein (GFP in the cultured cells (osteoblasts; NOS-1 cells. The transfection efficiency increased over 30% in the electroporation samples treated with D-glucosamine-supplemented buffer after one day. The membrane absorption of D-glucosamine is the primary mechanism of membrane stress induced by electric stress. This new function of D-glucosamine is useful and meaningful for developing more effective transformation procedures.

  1. Transfection of glioma cells with the neural-cell adhesion molecule NCAM

    DEFF Research Database (Denmark)

    Edvardsen, K; Pedersen, P H; Bjerkvig, R

    1994-01-01

    The tumor growth and the invasive capacity of a rat glioma cell line (BT4Cn) were studied after transfection with the human transmembrane 140-kDa isoform of the neural-cell adhesion molecule, NCAM. After s.c. injection, the NCAM-transfected cells showed a slower growth rate than the parent cell...

  2. Chitosan/Hyaluronic Acid Nanoparticles: Rational Design Revisited for RNA Delivery.

    Science.gov (United States)

    Lallana, Enrique; Rios de la Rosa, Julio M; Tirella, Annalisa; Pelliccia, Maria; Gennari, Arianna; Stratford, Ian J; Puri, Sanyogitta; Ashford, Marianne; Tirelli, Nicola

    2017-07-03

    Chitosan/hyaluronic acid (HA) nanoparticles can be used to deliver an RNA/DNA cargo to cells overexpressing HA receptors such as CD44. For these systems, unequivocal links have not been established yet between chitosan macromolecular (molecular weight; degree of deacetylation, i.e., charge density) and nanoparticle variables (complexation strength, i.e., stability; nucleic acid protection; internalization rate) on one hand, and transfection efficiency on the other hand. Here, we have focused on the role of avidity on transfection efficiency in the CD44-expressing HCT-116 as a cellular model; we have employed two differently sized payloads (a large luciferase-encoding mRNA and a much smaller anti-Luc siRNA), and a small library of chitosans (variable molecular weight and degree of deactylation). The RNA avidity for chitosan showed-as expected-an inverse relationship: higher avidity-higher polyplex stability-lower transfection efficiency. The avidity of chitosan for RNA appears to lead to opposite effects: higher avidity-higher polyplex stability but also higher transfection efficiency. Surprisingly, the best transfecting particles were those with the lowest propensity for RNA release, although this might be a misleading relationship: for example, the same macromolecular parameters that increase avidity can also boost chitosan's endosomolytic activity, with a strong enhancement in transfection. The performance of these nonviral vectors appears therefore difficult to predict simply on the basis of carrier- or payload-related variables, and a more holistic consideration of the journey of the nanoparticle, from cell uptake to cytosolic bioavailability of payload, is needed. It is also noteworthy that the nanoparticles used in this study showed optimal performance under slightly acidic conditions (pH 6.4), which is promising for applications in a tumoral extracellular environment. It is also worth pointing out that under these conditions we have for the first time

  3. The use of computerized video time lapse to study cell death in rat embryo cells transfected with c-ha-ras or c-myc

    International Nuclear Information System (INIS)

    Forrester, H.B.; Vidair, C.A.; Dewey, W.C.; Ling, C.C.

    1998-01-01

    Full text: Individual rat embryo fibroblasts that had been transfected with the c-myc (REC:myc) or c-Ha ras (REC:ras) oncogene were followed after irradiation using a computer video time lapse (CVTL) system in order to quantify the lethal events that resulted in loss of clonogenic survival after irradiation. By followed the cells for 2 to 3 generations before irradiation we were able to determine where they were in the cell cycle at the time of irradiation for cell cycle analysis. After irradiation, the individual cells and their progeny were followed in multiple fields for 5-6 days Then, pedigrees for individual irradiated cells were determined by noting the times of divisions fusions, and cell death. After X-irradiation, the clonogenic survival values for these two cell lines are similar. However, by using computerized video time lapse (CVTL) to follow individual cells we found that the loss of clonogenic survival was due to two different processes, cell death and a senescent-like process. The loss of clonogenic survival of x-irradiated (9.5 and 4 Gy) REC:myc cells was attributed almost entirely to the cells dying by apoptosis (∼99 and 90%). In contrast, approximately 60% of the x-irradiated (9.5 Gy) non-clonogenic REC:ras cells died by apoptosis (with a very small amount of necrosis), and the other 40% underwent a senescent-type process in which some of the cells and their progeny stopped dividing but remained as viable cells throughout 140 hours of observation. Both processes usually occurred after the cells had divided and continued to occur in the cells' progeny for up to five divisions after irradiation. The mode of cell death in the progeny of a non-clonogenic cell can be determined only by using CVTL and can not be determined by conventional clonogenic survival experiments. Also, only by following the individual cells and their progeny can the true amount of apoptosis be determined. The cumulative percentage of apoptosis scored in whole populations

  4. Environmental parameters influence non-viral transfection of human mesenchymal stem cells for tissue engineering applications

    Science.gov (United States)

    King, William J.; Kouris, Nicholas A.; Choi, Siyoung; Ogle, Brenda M.; Murphy, William L.

    2012-01-01

    Non-viral transfection is a promising technique which could be used to increase the therapeutic potential of stem cells. The purpose of this study was to explore practical culture parameters of relevance in potential human mesenchymal stem cell (hMSC) clinical and tissue engineering applications, including type of polycationic transfection reagent, N/P ratio and dose of polycation/pDNA polyplexes, cell passage number, cell density, and cell proliferation. The non-viral transfection efficiency was significantly influenced by N/P ratio, polyplex dose, cell density, and cell passage number. hMSC culture conditions that inhibited cell division also decreased transfection efficiency, suggesting that strategies to promote hMSC proliferation may be useful to enhance transfection efficiency in future tissue engineering studies. Non-viral transfection treatments influenced hMSC phenotype, including the expression level of the hMSC marker CD105, and the ability of hMSCs to differentiate down the osteogenic and adipogenic lineages. The parameters found here to promote hMSC transfection efficiency, minimize toxicity, and influence hMSC phenotype may be instructive in future non-viral transfection studies and tissue engineering applications. PMID:22277991

  5. Metallic nanoparticles in dielectrics: A comparative study

    KAUST Repository

    Agambayev, Agamyrat

    2017-10-25

    The Maxwell-Garnett method is used to predict the effective dielectric constant and the tangent loss of various composites consisting of a PVDF-TrFE-CFE-matrix and metallic microsphere fillers made of Cu, Ni, W, Zn, or Fe. Simulation results demonstrate that for small filler fraction values and at low frequencies, the electrical properties of the resulting composite do not depend on the conductivity of the filler. These findings show that composites fabricated using cheaper metal nanoparticle fillers are as effective as those fabricated using expensive ones.

  6. Magnetic nanoparticles studied by small angle X-ray scattering

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Cristiano Luis Pinto [Universidade de Sao Paulo (IF/USP), SP (Brazil). Inst. de Fisica. Grupo de Fluidos Complexos; Antonel, Soledad; Negri, Martin [Universidad de Buenos Aires (UBA) (Argentina). Facultad de Ciencias Exactas y Naturales. Dept. de Quimica Inorganica, Analitica y Quimica Fisica

    2011-07-01

    Full text: Magnetic nanoparticles have attracted much attention in the past decades because of their potential applications in high-density magnetic recording, magnetic fluids, data storage, spin-tronics, solar cells, sensors and catalysis. Among the magnetic nanoparticles, cobalt ferrite (CoFe{sub 2}O{sub 4}) has been widely studied due to high electromagnetic performance, excellent chemical stability, mechanical hardness, and high cubic magnetocrystalline anisotropy. These properties make it a promising candidate for many applications in commercial electronics such as video, audio tapes, high-density digital recording media, and magnetic fluids. Other interesting application is the use of magnetic nanocompounds in the design of magneto elastomers. Magnetoelastomers are dispersions of magnetic particles into an elastomer polymer matrix. These materials are highly promising for applications in the development of sensors and actuators, mainly because of the possibility to optimize the quality parameters of the devices by systematically changing the chemical nature of both the inorganic particles and the organic polymeric matrix, with the consequent modification of the magnetic, electric and elastic properties. Moreover, nanoparticles of cobalt-iron oxides (cobalt ferrite, CoFe{sub 2}O{sub 4}) appears as very interesting compounds for magnetoelasticity, not only because present magnetic anisotropy, moderate-high magnetization and high coercitivity at room temperature, but also because the possibility to modulate its magnetic properties by chemical synthesis, that is by synthesizing nanoparticles of different sizes having thus not only different magnetic parameters but also different magnetic behavior (superparamagnetism or ferromagnetism). That means that most of the magnetic properties of CoFe{sub 2}O{sub 4} ferrite strongly depend on the size and shape of the nanoparticles, which are closely related to the method of preparation. On the other hand, nickel

  7. Magnetic nanoparticles studied by small angle X-ray scattering

    International Nuclear Information System (INIS)

    Oliveira, Cristiano Luis Pinto; Antonel, Soledad; Negri, Martin

    2011-01-01

    Full text: Magnetic nanoparticles have attracted much attention in the past decades because of their potential applications in high-density magnetic recording, magnetic fluids, data storage, spin-tronics, solar cells, sensors and catalysis. Among the magnetic nanoparticles, cobalt ferrite (CoFe 2 O 4 ) has been widely studied due to high electromagnetic performance, excellent chemical stability, mechanical hardness, and high cubic magnetocrystalline anisotropy. These properties make it a promising candidate for many applications in commercial electronics such as video, audio tapes, high-density digital recording media, and magnetic fluids. Other interesting application is the use of magnetic nanocompounds in the design of magneto elastomers. Magnetoelastomers are dispersions of magnetic particles into an elastomer polymer matrix. These materials are highly promising for applications in the development of sensors and actuators, mainly because of the possibility to optimize the quality parameters of the devices by systematically changing the chemical nature of both the inorganic particles and the organic polymeric matrix, with the consequent modification of the magnetic, electric and elastic properties. Moreover, nanoparticles of cobalt-iron oxides (cobalt ferrite, CoFe 2 O 4 ) appears as very interesting compounds for magnetoelasticity, not only because present magnetic anisotropy, moderate-high magnetization and high coercitivity at room temperature, but also because the possibility to modulate its magnetic properties by chemical synthesis, that is by synthesizing nanoparticles of different sizes having thus not only different magnetic parameters but also different magnetic behavior (superparamagnetism or ferromagnetism). That means that most of the magnetic properties of CoFe 2 O 4 ferrite strongly depend on the size and shape of the nanoparticles, which are closely related to the method of preparation. On the other hand, nickel nanoparticles are very interesting

  8. A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells.

    Science.gov (United States)

    Homann, Stefanie; Hofmann, Christian; Gorin, Aleksandr M; Nguyen, Huy Cong Xuan; Huynh, Diana; Hamid, Phillip; Maithel, Neil; Yacoubian, Vahe; Mu, Wenli; Kossyvakis, Athanasios; Sen Roy, Shubhendu; Yang, Otto Orlean; Kelesidis, Theodoros

    2017-01-01

    Transfection is one of the most frequently used techniques in molecular biology that is also applicable for gene therapy studies in humans. One of the biggest challenges to investigate the protein function and interaction in gene therapy studies is to have reliable monospecific detection reagents, particularly antibodies, for all human gene products. Thus, a reliable method that can optimize transfection efficiency based on not only expression of the target protein of interest but also the uptake of the nucleic acid plasmid, can be an important tool in molecular biology. Here, we present a simple, rapid and robust flow cytometric method that can be used as a tool to optimize transfection efficiency at the single cell level while overcoming limitations of prior established methods that quantify transfection efficiency. By using optimized ratios of transfection reagent and a nucleic acid (DNA or RNA) vector directly labeled with a fluorochrome, this method can be used as a tool to simultaneously quantify cellular toxicity of different transfection reagents, the amount of nucleic acid plasmid that cells have taken up during transfection as well as the amount of the encoded expressed protein. Finally, we demonstrate that this method is reproducible, can be standardized and can reliably and rapidly quantify transfection efficiency, reducing assay costs and increasing throughput while increasing data robustness.

  9. Spatial and Temporal Control of Cavitation Allows High In Vitro Transfection Efficiency in the Absence of Transfection Reagents or Contrast Agents.

    Science.gov (United States)

    Chettab, Kamel; Roux, Stéphanie; Mathé, Doriane; Cros-Perrial, Emeline; Lafond, Maxime; Lafon, Cyril; Dumontet, Charles; Mestas, Jean-Louis

    2015-01-01

    Sonoporation using low-frequency high-pressure ultrasound (US) is a non-viral approach for in vitro and in vivo gene delivery. In this study, we developed a new sonoporation device designed for spatial and temporal control of ultrasound cavitation. The regulation system incorporated in the device allowed a real-time control of the cavitation level during sonoporation. This device was evaluated for the in vitro transfection efficiency of a plasmid coding for Green Fluorescent Protein (pEGFP-C1) in adherent and non-adherent cell lines. The transfection efficiency of the device was compared to those observed with lipofection and nucleofection methods. In both adherent and non-adherent cell lines, the sonoporation device allowed high rate of transfection of pEGFP-C1 (40-80%), as determined by flow cytometry analysis of GFP expression, along with a low rate of mortality assessed by propidium iodide staining. The transfection efficiency and toxicity of sonoporation on the non-adherent cell lines Jurkat and K562 were similar to those of nucleofection, while these two cell lines were resistant to transfection by lipofection. Moreover, sonoporation was used to produce three stably transfected human lymphoma and leukemia lines. Significant transfection efficiency was also observed in two fresh samples of human acute myeloid leukemia cells. In conclusion, we developed a user-friendly and cost-effective ultrasound device, well adapted for routine in vitro high-yield transfection experiments and which does not require the use of any transfection reagent or gas micro-bubbles.

  10. Transfected parvalbumin alters calcium homeostasis in teratocarcinoma PCC7 cells

    DEFF Research Database (Denmark)

    Müller, B K; Kabos, P; Belhage, B

    1996-01-01

    transfected. Parvalbumin-transfected and mock-transfected cells were loaded with the calcium indicator fura-2 and were exposed, in the same dish, to different concentrations of the calcium ionophore A23187 or to KCI. The results show that parvalbumin-transfected PCC7 cells had much better calcium buffering...

  11. EXAFS Studies of Palladium Nanoparticles: Size Control and Hydrogenation

    Science.gov (United States)

    Harris, T.; Soussan, L.; Isseroff, R.; Sun, Y.; Rafailovich, M. H.; Frenkel, A. I.

    2007-02-01

    We synthesized several samples of thiol-stabilized Pd nanoparticles by controlling their size with the palladium/thiol ratio from 5:1 to 1:3. Size dependence on the Pd/thiol ratio was verified by comparing the Pd-S and Pd-Pd coordination numbers. The average size of free-standing nanoparticles decreased as the thiol concentration increased and stabilized at the 1:2 value of palladium/thiol ratio. In addition to the free-standing nanoparticles, we analyzed their thin films by depositing them as monolayers on Kapton film using Langmuir-Blodgett technique. We obtained that the structure of the nanoparticles was preserved, within the accuracy of the EXAFS measurement, after preparing Langmuir films. We also studied the effect of hydrogenation on the both type of the nanoparticles (the free standing ones and those in the monolayers). In both cases, exposure to 4 atm H2 pressure resulted in the elongation of the average Pd-Pd distance. The larger change (0.06±0.03 Å, relative to the non-hydrogenated films) was observed in the smaller clusters (with 1:1 Pd thiol ratio), and the smaller change (0.03±0.01 Å) — in the larger clusters (with 3:1 ratio). Such bond length expansion is attributed to the defects caused by H2 adsorption.

  12. In vitro toxicological nanoparticle studies under flow exposure

    Energy Technology Data Exchange (ETDEWEB)

    Sambale, Franziska, E-mail: sambale@iftc.uni-hannover.de; Stahl, Frank; Bahnemann, Detlef; Scheper, Thomas [Gottfried Wilhelm Leibniz University Hanover, Institute for Technical Chemistry (Germany)

    2015-07-15

    The use of nanoparticles is becoming increasingly common in industry and everyday objects. Thus, extensive risk management concerning the potential health risk of nanoparticles is important. Currently, in vitro nanoparticle testing is mainly performed under static culture conditions without any shear stress. However, shear stress is an important biomechanical parameter. Therefore, in this study, a defined physiological flow to different mammalian cell lines such as A549 cells and NIH-3T3 cells has been applied. The effects of zinc oxide and titanium dioxide nanoparticles (TiO{sub 2}-NP), respectively, were investigated under both static and dynamic conditions. Cell viability, cell morphology, and adhesion were proven and compared to the static cell culture. Flow exposure had an impact on the cellular morphology of the cells. NIH-3T3 cells were elongated in the direction of flow and A549 cells exhibited vesicles inside the cells. Zinc oxide nanoparticles reduced the cell viability in the static and in the dynamic culture; however, the dynamic cultures were more sensitive. In the static culture and in the dynamic culture, TiO{sub 2}-NP did not affect cell viability. In conclusion, dynamic culture conditions are important for further in vitro investigations and provide more relevant results than static culture conditions.

  13. A Study on Gamma Irradiation Synthesis of Copper Nanoparticles

    International Nuclear Information System (INIS)

    Ahmad, Shahrul Izwan Bin; Ahmad, Md. Soot Bin Hj.; Radiman, Shahidan Bin

    2009-01-01

    A study on the effect of gamma radiation dose and dose rate on the yield of copper nanoparticles produced had been done. Its objective is to show the relationship between the absorbed doses with the yield of production. The copper sulphate solution is prepared with addition of ethanol as radical scavenger. Then the solution is bubbled with nitrogen for before being irradiated at different absorbed dose and dose rate. There are five different dose rates being used in this experiment. Atomic absorption spectroscopy (AAS) was used to detect directly the quantity of copper nanoparticles produced. The AAS results show positive linear relationship between the yields of copper nanoparticles with increasing absorbed dose. Yield of production show independency with dose rate at every absorbed dose. AAS result is supported with UV-Vis analysis data on the supernatant from irradiated products. Transmission electron microscope (TEM) confirms the existence of copper nanoparticles in all samples that being irradiated at absorbed dose of 100 kGy. The size of nanoparticles is range from 2 to 10 nm. Peak from the XRD analysis show the existence of pure copper.

  14. Optimized PEI-based Transfection Method for Transient Transfection and Lentiviral Production.

    Science.gov (United States)

    Yang, Shaozhe; Zhou, Xiaoling; Li, Rongxiang; Fu, Xiuhong; Sun, Pingnan

    2017-09-14

    Polyethyleneimine (PEI), a cationic polymer vehicle, forms a complex with DNA which then can carry anionic nucleic acids into eukaryotic cells. PEI-based transfection is widely used for transient transfection of plasmid DNA. The efficiency of PEI-based transfection is affected by numerous factors, including the way the PEI/DNA complex is prepared, the ratio of PEI to DNA, the concentration of DNA, the storage conditions of PEI solutions, and more. Considering the major influencing factors, PEI-based transfection has been optimized to improve its efficiency, reproducibility, and consistency. This protocol outlines the steps for ordinary transient transfection and lentiviral production using PEI. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley and Sons, Inc.

  15. Cationic Polybutyl Cyanoacrylate Nanoparticles for DNA Delivery

    Directory of Open Access Journals (Sweden)

    Jinghua Duan

    2009-01-01

    Full Text Available To enhance the intracellular delivery potential of plasmid DNA using nonviral vectors, we used polybutyl cyanoacrylate (PBCA and chitosan to prepare PBCA nanoparticles (NPs by emulsion polymerization and prepared NP/DNA complexes through the complex coacervation of nanoparticles with the DNA. The object of our work is to evaluate the characterization and transfection efficiency of PBCA-NPs. The NPs have a zeta potential of 25.53 mV at pH 7.4 and size about 200 nm. Electrophoretic analysis suggested that the NPs with positive charges could protect the DNA from nuclease degradation and cell viability assay showed that the NPs exhibit a low cytotoxicity to human hepatocellular carcinoma (HepG2 cells. Qualitative and quantitative analysis of transfection in HepG2 cells by the nanoparticles carrying plasmid DNA encoding for enhanced green fluorescent protein (EGFP-N1 was done by digital fluorescence imaging microscopy system and fluorescence-activated cell sorting (FACS. Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours. Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation. The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery.

  16. A High-Throughput Microfluidic Platform for Mammalian Cell Transfection and Culturing

    Science.gov (United States)

    Woodruff, Kristina; Maerkl, Sebastian J.

    2016-01-01

    Mammalian synthetic biology could be augmented through the development of high-throughput microfluidic systems that integrate cellular transfection, culturing, and imaging. We created a microfluidic chip that cultures cells and implements 280 independent transfections at up to 99% efficiency. The chip can perform co-transfections, in which the number of cells expressing each protein and the average protein expression level can be precisely tuned as a function of input DNA concentration and synthetic gene circuits can be optimized on chip. We co-transfected four plasmids to test a histidine kinase signaling pathway and mapped the dose dependence of this network on the level of one of its constituents. The chip is readily integrated with high-content imaging, enabling the evaluation of cellular behavior and protein expression dynamics over time. These features make the transfection chip applicable to high-throughput mammalian protein and synthetic biology studies. PMID:27030663

  17. Sum Frequency Generation Studies of Hydrogenation Reactions on Platinum Nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Krier, James M. [Univ. of California, Berkeley, CA (United States)

    2013-08-31

    Sum Frequency Generation (SFG) vibrational spectroscopy is used to characterize intermediate species of hydrogenation reactions on the surface of platinum nanoparticle catalysts. In contrast to other spectroscopy techniques which operate in ultra-high vacuum or probe surface species after reaction, SFG collects information under normal conditions as the reaction is taking place. Several systems have been studied previously using SFG on single crystals, notably alkene hydrogenation on Pt(111). In this thesis, many aspects of SFG experiments on colloidal nanoparticles are explored for the first time. To address spectral interference by the capping agent (PVP), three procedures are proposed: UV cleaning, H2 induced disordering and calcination (core-shell nanoparticles). UV cleaning and calcination physically destroy organic capping while disordering reduces SFG signal through a reversible structural change by PVP.

  18. Critical Parametric Study on Final Size of Magnetite Nanoparticles

    Science.gov (United States)

    Yusoff, A. H. M.; Salimi, M. N.; Jamlos, M. F.

    2018-03-01

    The great performance of magnetite nanoparticle in varsity field are mainly depended on their size since size determine the saturation magnetisation and also the phase purity. Magnetite nanoparticles were prepared using a simple co-precipitation method in order to study the influence of synthesis condition on the final size. Variable parameters include stirring rate, reaction temperature and pH of the solution can finely tuned the size of the resulting nanoparticles. Generally, any increase in these parameters had a gently reduction on particle size. But, the size was promoted to increase back at certain point due to the specific reason. Nucleation and growth processes are involved to clarify the impact of synthesis condition on the particle sizes. The result obtained give the correct conditions for pure magnetite synthesis at nanoscale size of dimensions less than 100 nm.

  19. A study of multifunctionalization of fluorescent nanoparticle probes

    Science.gov (United States)

    Wong, Hon Tung

    In this project, structural, luminescent and magnetic properties of multifunctional trivalent rare-earth ion (RE3+)-doped down- and up-converting nanoparticles with intrinsic magnetism and luminescence have been investigated. Syntheses, characterizations and surface modifications of multifunctional RE3+-doped nanoparticles have been carried out. All the multifunctional RE3+-doped down- and up-converting nanophosphors were synthesized using a facile hydrothermal method and a novel solution-based method. A variety of characterization techniques have been performed on the obtained phosphors, including X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), photoluminescent spectroscopy (PL), photoluminescent excitation spectroscopy (PLE), lifetime measurement, laser power measurement, vibrating sample magnetometer (VSM), Raman spectroscopy and confocal microscopy. An overview of the studies performed in this project is as follows. Different oxide and fluoride hosts have been synthesized and studied, including Gd 2O3, GdF3, YF3, NaYF4, and the seldom reported KGdF4. Different RE3+ ions have been doped into the above mentioned hosts for the realization of multifunctional nanoparticles with down- and up-conversion. For downconverting phosphors, Eu3+, Tb3+ and Tm3+ ions have been doped to obtain red, green, and blue emission. For upconverting phosphors, Tm3+/Yb3+ and Er3+/Yb 3+ ions have been doped to attain near-infrared to near-infrared and near-infrared to visible upconversion. The effects of hosts on the structural, luminescent and magnetic properties have been systemically studied. It is found that both hosts and dopants concentrations play a major role in determining the spectral features and the luminescent emission intensity. Particularly, nearly pure near-infrared to near-infrared upconversion is obtained in a specific GdF3 host. Characterizations of magnetic properties

  20. A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots

    International Nuclear Information System (INIS)

    Zhang, Hai-Li; Zhang, Ming-Zhen; Li, Xiang-Yong; Wan, Min; Li, Yong-Qiang; Zhang, Rong-Ying; Zhao, Yuan-Di

    2012-01-01

    Highlights: ► An easy and direct way to prepare QDs–DNA complexes was developed. ► Surface charge of QDs was tuned with different ratio of amino and glycolate. ► Transfection efficiency was dependent on the surface zeta potentials of QDs. ► Cellular toxicity of this gene vectors is much lower than commercial liposome. ► Whole intracellular behavior of QDs–DNA complexes can be monitored in real time. -- Abstract: Nanoparticle carrier has been developed by combining water-soluble quantum dots and plasmid DNA expressed enhanced green fluorescent protein (EGFP) in a convenient and direct way. First the QDs with different surface charges were obtained by coating with amino and carboxyl terminals at different ratios. Then plasmid DNA was conjugated to QDs via electrostatic interaction. The resultant QDs–DNA complexes showed enhanced resistance to DNase I digestion. The following transfection experiments demonstrated that the transfection efficiency was dependent on the surface charges on QDs. The real time imaging of the transfection process showed that the nanoparticles experienced binding, penetrating the cell membrane and entering cytoplasm in the first 6 h of transfection. The green fluorescence of EGFP began to appear after 18 h transfection and plasmid DNA was fully expressed in the following 6 h. This new QDs–DNA platform showed great potential as new gene delivery carrier.

  1. In vitro placental model optimization for nanoparticle transport studies

    Directory of Open Access Journals (Sweden)

    Cartwright L

    2012-01-01

    Full Text Available Laura Cartwright1, Marie Sønnegaard Poulsen2, Hanne Mørck Nielsen3, Giulio Pojana4, Lisbeth E Knudsen2, Margaret Saunders1, Erik Rytting2,51Bristol Initiative for Research of Child Health (BIRCH, Biophysics Research Unit, St Michael's Hospital, UH Bristol NHS Foundation Trust, Bristol, UK; 2University of Copenhagen, Faculty of Health Sciences, Department of Public Health, 3University of Copenhagen, Faculty of Pharmaceutical Sciences, Department of Pharmaceutics and Analytical Chemistry, Copenhagen, Denmark; 4Department of Environmental Sciences, Informatics and Statistics, University Ca' Foscari Venice, Venice, Italy; 5Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, Texas, USABackground: Advances in biomedical nanotechnology raise hopes in patient populations but may also raise questions regarding biodistribution and biocompatibility, especially during pregnancy. Special consideration must be given to the placenta as a biological barrier because a pregnant woman's exposure to nanoparticles could have significant effects on the fetus developing in the womb. Therefore, the purpose of this study is to optimize an in vitro model for characterizing the transport of nanoparticles across human placental trophoblast cells.Methods: The growth of BeWo (clone b30 human placental choriocarcinoma cells for nanoparticle transport studies was characterized in terms of optimized Transwell® insert type and pore size, the investigation of barrier properties by transmission electron microscopy, tight junction staining, transepithelial electrical resistance, and fluorescein sodium transport. Following the determination of nontoxic concentrations of fluorescent polystyrene nanoparticles, the cellular uptake and transport of 50 nm and 100 nm diameter particles was measured using the in vitro BeWo cell model.Results: Particle size measurements, fluorescence readings, and confocal microscopy indicated both cellular uptake of

  2. Gene therapy for C-26 colon cancer using heparin-polyethyleneimine nanoparticle-mediated survivin T34A

    Science.gov (United States)

    Zhang, Ling; Gao, Xiang; Men, Ke; Wang, BiLan; Zhang, Shuang; Qiu, Jinfeng; Huang, Meijuan; Gou, MaLing; Huang, Ning; Qian, ZhiYong; Zhao, Xia; Wei, YuQuan

    2011-01-01

    Background Gene therapy provides a novel method for the prevention and treatment of cancer, but the clinical application of gene therapy is restricted, mainly because of the absence of an efficient and safe gene delivery system. Recently, we developed a novel nonviral gene carrier, ie, heparin-polyethyleneimine (HPEI) nanoparticles for this purpose. Methods and results HPEI nanoparticles were used to deliver plasmid-expressing mouse survivin-T34A (ms-T34A) to treat C-26 carcinoma in vitro and in vivo. According to the in vitro studies, HPEI nanoparticles could efficiently transfect the pGFP report gene into C-26 cells, with a transfection efficiency of 30.5% ± 2%. Moreover, HPEI nanoparticle-mediated ms-T34A could efficiently inhibit the proliferation of C-26 cells by induction of apoptosis in vitro. Based on the in vivo studies, HPEI nanoparticles could transfect the Lac-Z report gene into C-26 cells in vivo. Intratumoral injection of HPEI nanoparticle-mediated ms-T34A significantly inhibited growth of subcutaneous C-26 carcinoma in vivo by induction of apoptosis and inhibition of angiogenesis. Conclusion This research suggests that HPEI nanoparticle-mediated ms-T34A may have a promising role in C-26 colon carcinoma therapy. PMID:22072877

  3. Surface oxidation of cobalt nanoparticles studied by Mossbauer spectroscopy

    DEFF Research Database (Denmark)

    Bødker, Franz; Mørup, Steen; Charles, S.W.

    1999-01-01

    The surface oxide formed on cobalt nanoparticles has been studied by Mossbauer emission spectroscopy. Exposure of the cobalt particles to oxygen at room temperature was found to result in the formation of a relatively well-ordered surface oxide with Mossbauer parameters similar to those of CoO....

  4. nanoparticles

    Science.gov (United States)

    Andreu-Cabedo, Patricia; Mondragon, Rosa; Hernandez, Leonor; Martinez-Cuenca, Raul; Cabedo, Luis; Julia, J. Enrique

    2014-10-01

    Thermal energy storage (TES) is extremely important in concentrated solar power (CSP) plants since it represents the main difference and advantage of CSP plants with respect to other renewable energy sources such as wind, photovoltaic, etc. CSP represents a low-carbon emission renewable source of energy, and TES allows CSP plants to have energy availability and dispatchability using available industrial technologies. Molten salts are used in CSP plants as a TES material because of their high operational temperature and stability of up to 500°C. Their main drawbacks are their relative poor thermal properties and energy storage density. A simple cost-effective way to improve thermal properties of fluids is to dope them with nanoparticles, thus obtaining the so-called salt-based nanofluids. In this work, solar salt used in CSP plants (60% NaNO3 + 40% KNO3) was doped with silica nanoparticles at different solid mass concentrations (from 0.5% to 2%). Specific heat was measured by means of differential scanning calorimetry (DSC). A maximum increase of 25.03% was found at an optimal concentration of 1 wt.% of nanoparticles. The size distribution of nanoparticle clusters present in the salt at each concentration was evaluated by means of scanning electron microscopy (SEM) and image processing, as well as by means of dynamic light scattering (DLS). The cluster size and the specific surface available depended on the solid content, and a relationship between the specific heat increment and the available particle surface area was obtained. It was proved that the mechanism involved in the specific heat increment is based on a surface phenomenon. Stability of samples was tested for several thermal cycles and thermogravimetric analysis at high temperature was carried out, the samples being stable.

  5. Fibronectin enhances transfection of Staphylococcus aureus.

    OpenAIRE

    Thompson, N E; Bergdoll, M S; Pattee, P A

    1985-01-01

    The factor in normal sera primarily responsible for the enhancement of transfection (and transformation) of Staphylococcus aureus was identified as fibronectin. Serum samples which were depleted of fibronectin by affinity chromatography showed a marked decrease in enhancing activity. Fibronectin isolated from sera of several animal species demonstrated enhancing activity.

  6. Efficient gene delivery to human umbilical cord mesenchymal stem cells by cationized Porphyra yezoensis polysaccharide nanoparticles.

    Science.gov (United States)

    Yu, Qingtong; Cao, Jin; Chen, Baoding; Deng, Wenwen; Cao, Xia; Chen, Jingjing; Wang, Yan; Wang, Shicheng; Yu, Jiangnan; Xu, Ximing; Gao, Xiangdong

    2015-01-01

    This study centered on an innovative application of Porphyra yezoensis polysaccharide (PPS) with cationic modification as a safe and efficient nonviral gene vector to deliver a plasmid encoding human Wnt3a (pWnt3a) into human umbilical cord mesenchymal stem cells (HUMSCs). After modification with branched low-molecular-weight (1,200 Da) polyethylenimine, the cationized PPS (CPPS) was combined with pWnt3a to form spherical nanoscale particles (CPPS-pWnt3a nanoparticles). Particle size and distribution indicated that the CPPS-pWnt3a nanoparticles at a CPPS:pWnt3a weight ratio of 40:1 might be a potential candidate for DNA plasmid transfection. A cytotoxicity assay demonstrated that the nanoparticles prepared at a CPPS:pWnt3a weight ratio of 40:1 were nontoxic to HUMSCs compared to those of Lipofectamine 2000 and polyethylenimine (25 kDa). These nanoparticles were further transfected to HUMSCs. Western blotting demonstrated that the nanoparticles (CPPS:pWnt3a weight ratio 40:1) had the greatest transfection efficiency in HUMSCs, which was significantly higher than that of Lipofectamine 2000; however, when the CPPS:pWnt3a weight ratio was increased to 80:1, the nanoparticle-treated group showed no obvious improvement in translation efficiency over Lipofectamine 2000. Therefore, CPPS, a novel cationic polysaccharide derived from P. yezoensis, could be developed into a safe, efficient, nonviral gene vector in a gene-delivery system.

  7. Feasibility and Limits of Magnetically Labeling Primary Cultured Rat T Cells with Ferumoxides Coupled with Commonly Used Transfection Agents

    Directory of Open Access Journals (Sweden)

    Cedric Berger

    2006-04-01

    Full Text Available Visualization and quantification of inflammatory processes is of high importance for early diagnosis of a multitude of diseases. Magnetic resonance imaging (MRI using iron oxide (FeO nanoparticles as contrast agents allows the study of macrophage infiltration during inflammation in a variety of tissues. Macrophages are effectors of the immune response, their appearance being orchestrated by activated T lymphocytes. Therefore, tracking of labeled T lymphocytes, which initiate the immune process, should enable earlier detection of tissue inflammation. In this study, we investigate the feasibility of specifically labeling harvested T cells by using dextran-coated FeO nanoparticles and commonly available transfection agents (TAs. Physicochemical properties of the newly formed FeO/TA vesicles were determined as well as their cell toxicity and their T cell activation potential. The labeling efficiency of each FeO/TA combination was evaluated by measuring the transverse MRI relaxation rate R2 by X-ray spectroscopy and magnetic selection. Toxicity and labeling efficacy differed significantly among TAs. The best results were achieved by using polyamine TAs and in particular by using poly-l-lysine at a concentration of 1.5 µg/mL administered in combination with 22.5 µg iron/mL. By using this protocol, up to 60% of harvested T cells could be labeled. Microscopic investigation revealed FeO/TA nanoparticles not only localized within the cytoplasma of the cells but also sticking to the outer membrane surface.

  8. Study of optical phonon modes of CdS nanoparticles using Raman ...

    Indian Academy of Sciences (India)

    In this paper we report the study of optical phonon modes of nanoparticles of CdS using Raman spectroscopy. Nanoparticle sample for the present study was synthesized through chemical precipitation technique. The CdS nanoparticles were then subjected to heat treatment at low temperature (150°C) for extended time ...

  9. Uptake of DNA by cancer cells without a transfection reagent.

    Science.gov (United States)

    Kong, Yanping; Zhang, Xianbo; Zhao, Yongliang; Xue, Yanfang; Zhang, Ye

    2017-01-21

    Cancer cells exhibit elevated levels of glucose uptake and may obtain pre-formed, diet-derived fatty acids from the bloodstream to boost their rapid growth; they may also use nucleic acid from their microenvironment. The study of processing nucleic acid by cancer cells will help improve the understanding of the metabolism of cancer. DNA is commonly packaged into a viral or lipid particle to be transferred into cells; this process is called transfection in laboratory. Cancer cells are known for having gene mutations and the evolving ability of endocytosis. Their uptake of DNAs might be different from normal cells; they may take in DNAs directly from the environment. In this report, we studied the uptake of DNAs in cancer cells without a transfection reagent. A group of DNA fragments were prepared with PCR and labeled with isotope phosphorous-32 to test their uptake by Huh 7 (liver cancer) and THLE3 (normal liver cells) after incubation overnight by counting radioactivity of the cells' genomic DNA. Multiple cell lines including breast cancer and lung cancer were tested with the same method. DNA molecules were also labeled with fluorescence to test the location in the cells using a kit of "label it fluorescence in situ hybridization (FISH)" from Mirus (USA). The data demonstrated that hepatocellular carcinoma cells possess the ability to take in large DNA fragments directly without a transfection reagent whereas normal liver cells cannot. Huh7 and MDA-MB231 cells displayed a significantly higher Rhodamine density in the cytoplasmic phagosomes and this suggests that the mechanism of uptake of large DNA by cancer cells is likely endocytosis. The efficacy of uptake is related to the DNA's size. Some cell lines of lung cancer and breast cancer also showed similar uptake of DNA. In the present study, we have revealed the evidence that some cancer cells, but not nontumorigenic cells, can take DNA fragments directly from the environment without the aid of the transfecting

  10. DNA uptake, intracellular trafficking and gene transfection after ultrasound exposure.

    Science.gov (United States)

    Liu, Ying; Yan, Jing; Santangelo, Philip J; Prausnitz, Mark R

    2016-07-28

    Ultrasound has been studied as a promising tool for intracellular gene delivery. In this work, we studied gene transfection of a human prostate cancer cell line exposed to megahertz pulsed ultrasound in the presence of contrast agent and assessed the efficiency of fluorescently labelled DNA delivery into cell nuclei, which is necessary for gene transfection. At the sonication conditions studied, ~30% of cells showed DNA uptake 30min after sonication, but that fraction decreased over time to ~10% of cells after 24h. Most cells containing DNA had DNA in their nuclei, but the amount varied significantly. Transfection efficiency peaked at ~10% at 8h post sonication. Among those cells containing DNA, ~30% of DNA was localized in the cell nuclei, ~30% was in autophagosomes/autophagolysosomes and the remainder was "free" in the cytoplasm 30min after sonication. At later times up to 24h, ~30% of DNA continued to be found in the nuclei and most or all of the rest of the DNA was in autophagosomes/autophagolysosomes. These results demonstrate that ultrasound can deliver DNA into cell nuclei shortly after sonication and that the rest of the DNA can be cleared by autophagosomes/autophagolysosomes. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Simulation of micro/nano electroporation for cell transfection

    Science.gov (United States)

    Zhang, Guocheng; Fan, Na; Jiang, Hai; Guo, Jian; Peng, Bei

    2018-03-01

    The 3D micro/nano electroporation for transfection has become a powerful biological cell research technique with the development of micro-nano manufacturing technology. The micro channels connected the cells with transfection reagents on the chip were important to the transmemnbrane potentical, which directly influences the electroporation efficiency. In this study, a two-dimensional model for electroporation of cells was designed to address the effects of channels’ sizes and number on transmembrane potential. The simulation results indicated that the transmembrane potential increased with increasing size of channels’ entrances. Moreover, compared with single channel entrance, the transmembrane potential was higher when the cells located at multiple channels entrances. These results suggest that it IS required to develop higher micro manufacturing technology to create channels as we expected size.

  12. Silicalite nanoparticles that promote transgene expression

    International Nuclear Information System (INIS)

    Pearce, Megan E; Mai, Hoang Q; Salem, Aliasger K; Lee, Namhoon; Larsen, Sarah C

    2008-01-01

    Here, we report on a new zeolite-based silicalite nanoparticle that can enhance the transfection efficiencies generated by poly ethylene imine-plasmid DNA (PEI-pDNA) complexes via a sedimentation mechanism and can enhance the transfection efficiencies of pDNA alone when surface functionalized with amine groups. The silicalite nanoparticles have a mean size of 55 nm. Functionalizing the silicalite nanoparticles with amine groups results in a clear transition in zeta potential from -25.9 ± 2.3 mV (pH 7.4) for unfunctionalized silicalite nanoparticles to 4.9 ± 0.7 mV (pH 7.4) for amine functionalized silicalite nanoparticles. We identify that silicalite nanoparticles used to promote non-viral vector acceleration to the cell surface are found in acidic vesicles or the cytoplasm but not the nucleus. An MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay showed that the silicalite nanoparticles were non-toxic at the concentrations tested for transfection. We show that surface functionalization of silicalite nanoparticles with amine groups results in a significant (230%) increase in transfection efficiency of pDNA when compared to unfunctionalized silicalite nanoparticles. Silicalite nanoparticles enhanced pDNA-PEI induced transfection of human embryonic kidney (HEK-293) cells by over 150%

  13. Dual-degradable disulfide-containing PEI–Pluronic/DNA polyplexes: transfection efficiency and balancing protection and DNA release

    Directory of Open Access Journals (Sweden)

    Zhang L

    2013-09-01

    Full Text Available Lifen Zhang,* Zhenzhen Chen,* Yanfeng LiState Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metals Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology, Lanzhou University, Lanzhou, People's Republic of China*These authors contributed equally to this workAbstract: Polymeric gene-delivery vectors to achieve lack of toxicity and a balance between protection and DNA release remains a formidable challenge. Incorporating intracellular environment-responsive degradable bonds is an appreciable step toward developing safer transfection agents. In this study, novel, dual-degradable polycation copolymers (Pluronic-diacrylate [PA]–polyethyleneimine [PEI]–SS were synthesized through the addition of low molecular weight (800 Da PEI cross-linked with SS (PEI-SS to PA. Three PA-PEI-SS copolymers (PA-PEI-SS1, 2, and 3 with different PEI-SS to Pluronic molar ratios were investigated and found to strongly condense plasmid DNA into positively charged nanoparticles with an average particle size of approximately 200 nm and to possess higher stability against DNase I digestion and sodium heparin. Disulfide and ester bonds of the copolymers were susceptible to intracellular redox conditions. In vitro experiments demonstrated that the PA-PEI-SS copolymers had significantly lower cytotoxicity and higher transfection efficiency in both BGC-823 and 293T cell lines than the controls of degradable PEI-SS and nondegradable 25 kDa PEI. Transfection activity was influenced by the PEI-SS content in the polymers and PA-PEI-SS1 showed the highest efficiency of the three copolymers. These studies suggest that these dual-degradable copolymers could be used as potential biocompatible gene delivery carriers.Keywords: Pluronic, PEI, gene vector, dual-degradable, disulfide-containing linker

  14. A comprehensive high-throughput FTIR spectroscopy-based method for evaluating the transfection event: estimating the transfection efficiency and extracting associated metabolic responses.

    Science.gov (United States)

    Rosa, Filipa; Sales, Kevin C; Cunha, Bernardo R; Couto, Andreia; Lopes, Marta B; Calado, Cecília R C

    2015-10-01

    Reporter genes are routinely used in every laboratory for molecular and cellular biology for studying heterologous gene expression and general cellular biological mechanisms, such as transfection processes. Although well characterized and broadly implemented, reporter genes present serious limitations, either by involving time-consuming procedures or by presenting possible side effects on the expression of the heterologous gene or even in the general cellular metabolism. Fourier transform mid-infrared (FT-MIR) spectroscopy was evaluated to simultaneously analyze in a rapid (minutes) and high-throughput mode (using 96-wells microplates), the transfection efficiency, and the effect of the transfection process on the host cell biochemical composition and metabolism. Semi-adherent HEK and adherent AGS cell lines, transfected with the plasmid pVAX-GFP using Lipofectamine, were used as model systems. Good partial least squares (PLS) models were built to estimate the transfection efficiency, either considering each cell line independently (R (2) ≥ 0.92; RMSECV ≤ 2 %) or simultaneously considering both cell lines (R (2) = 0.90; RMSECV = 2 %). Additionally, the effect of the transfection process on the HEK cell biochemical and metabolic features could be evaluated directly from the FT-IR spectra. Due to the high sensitivity of the technique, it was also possible to discriminate the effect of the transfection process from the transfection reagent on KEK cells, e.g., by the analysis of spectral biomarkers and biochemical and metabolic features. The present results are far beyond what any reporter gene assay or other specific probe can offer for these purposes.

  15. Antigen-presenting cells transfected with Hsp65 messenger RNA fail to treat experimental tuberculosis

    International Nuclear Information System (INIS)

    Rocha, C.D.; Trombone, A.P.F.; Lorenzi, J.C.C.; Almeida, L.P.; Gembre, A.F.; Padilha, E.; Ramos, S.G.; Silva, C.L.; Coelho-Castelo, A.A.M.

    2012-01-01

    In the last several years, the use of dendritic cells has been studied as a therapeutic strategy against tumors. Dendritic cells can be pulsed with peptides or full-length protein, or they can be transfected with DNA or RNA. However, comparative studies suggest that transfecting dendritic cells with messenger RNA (mRNA) is superior to other antigen-loading techniques in generating immunocompetent dendritic cells. In the present study, we evaluated a new therapeutic strategy to fight tuberculosis using dendritic cells and macrophages transfected with Hsp65 mRNA. First, we demonstrated that antigen-presenting cells transfected with Hsp65 mRNA exhibit a higher level of expression of co-stimulatory molecules, suggesting that Hsp65 mRNA has immunostimulatory properties. We also demonstrated that spleen cells obtained from animals immunized with mock and Hsp65 mRNA-transfected dendritic cells were able to generate a mixed Th1/Th2 response with production not only of IFN-γ but also of IL-5 and IL-10. In contrast, cells recovered from mice immunized with Hsp65 mRNA-transfected macrophages were able to produce only IL-5. When mice were infected with Mycobacterium tuberculosis and treated with antigen-presenting cells transfected with Hsp65 mRNA (therapeutic immunization), we did not detect any decrease in the lung bacterial load or any preservation of the lung parenchyma, indicating the inability of transfected cells to confer curative effects against tuberculosis. In spite of the lack of therapeutic efficacy, this study reports for the first time the use of antigen-presenting cells transfected with mRNA in experimental tuberculosis

  16. Antigen-presenting cells transfected with Hsp65 messenger RNA fail to treat experimental tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, C.D.; Trombone, A.P.F.; Lorenzi, J.C.C.; Almeida, L.P.; Gembre, A.F.; Padilha, E. [Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Ramos, S.G. [Departamento de Patologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Silva, C.L.; Coelho-Castelo, A.A.M. [Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil)

    2012-09-21

    In the last several years, the use of dendritic cells has been studied as a therapeutic strategy against tumors. Dendritic cells can be pulsed with peptides or full-length protein, or they can be transfected with DNA or RNA. However, comparative studies suggest that transfecting dendritic cells with messenger RNA (mRNA) is superior to other antigen-loading techniques in generating immunocompetent dendritic cells. In the present study, we evaluated a new therapeutic strategy to fight tuberculosis using dendritic cells and macrophages transfected with Hsp65 mRNA. First, we demonstrated that antigen-presenting cells transfected with Hsp65 mRNA exhibit a higher level of expression of co-stimulatory molecules, suggesting that Hsp65 mRNA has immunostimulatory properties. We also demonstrated that spleen cells obtained from animals immunized with mock and Hsp65 mRNA-transfected dendritic cells were able to generate a mixed Th1/Th2 response with production not only of IFN-γ but also of IL-5 and IL-10. In contrast, cells recovered from mice immunized with Hsp65 mRNA-transfected macrophages were able to produce only IL-5. When mice were infected with Mycobacterium tuberculosis and treated with antigen-presenting cells transfected with Hsp65 mRNA (therapeutic immunization), we did not detect any decrease in the lung bacterial load or any preservation of the lung parenchyma, indicating the inability of transfected cells to confer curative effects against tuberculosis. In spite of the lack of therapeutic efficacy, this study reports for the first time the use of antigen-presenting cells transfected with mRNA in experimental tuberculosis.

  17. Optimization of transfection of green fluorescent protein in pursuing mesenchymal stem cells in vivo

    Directory of Open Access Journals (Sweden)

    Pınar Elçi

    2008-12-01

    Full Text Available OBJECTIVE: Green Fluorescent Protein (GFP has been used as a marker of gene expression and a single cell marker in living organisms in cell biology studies. The important areas that GFP is used are expression levels of different genes in different organisms by inserting GFP in these genes and as a marker in living cells. In this study, we tried to optimize transfection of mesenchymal stem cells, (MSCs used for regeneration of damaged tissues in animals, by GFP containing plasmid vector by which MSCs can be followed in vivo.METHODS: To this aim, phM-GFP plasmid vector carrying GFP gene and effectene transfection reagent were used. RESULTS: The data revealed that twice transfection of MSCs resulted in higher expression of GFP for longer times as compared to once transfected MSCs. On the other hand, leaving the chemical transfection agents in the medium induced apoptosis after a while. CONCLUSION: As a conclusion we suggest the transfection of MSCs twice with 48 hours interval and removal of transfection agents after 8 hours which removed toxic and apoptotic effects of the chemicals.

  18. Synergistic effect of electrical and chemical factors on endocytosis in micro-discharge plasma gene transfection

    Science.gov (United States)

    Jinno, M.; Ikeda, Y.; Motomura, H.; Isozaki, Y.; Kido, Y.; Satoh, S.

    2017-06-01

    We have developed a new micro-discharge plasma (MDP)-based gene transfection method, which transfers genes into cells with high efficiency and low cytotoxicity; however, the mechanism underlying the method is still unknown. Studies revealed that the N-acetylcysteine-mediated inhibition of reactive oxygen species (ROS) activity completely abolished gene transfer. In this study, we used laser-produced plasma to demonstrate that gene transfer does not occur in the absence of electrical factors. Our results show that both electrical and chemical factors are necessary for gene transfer inside cells by microplasma irradiation. This indicates that plasma-mediated gene transfection utilizes the synergy between electrical and chemical factors. The electric field threshold required for transfection was approximately 1 kV m-1 in our MDP system. This indicates that MDP irradiation supplies sufficient concentrations of ROS, and the stimulation intensity of the electric field determines the transfection efficiency in our system. Gene transfer by plasma irradiation depends mainly on endocytosis, which accounts for at least 80% of the transfer, and clathrin-mediated endocytosis is a dominant endocytosis. In plasma-mediated gene transfection, alterations in electrical and chemical factors can independently regulate plasmid DNA adhesion and triggering of endocytosis, respectively. This implies that plasma characteristics can be adjusted according to target cell requirements, and the transfection process can be optimized with minimum damage to cells and maximum efficiency. This may explain how MDP simultaneously achieves high transfection efficiency with minimal cell damage.

  19. Photoluminescence and conductivity studies of anthracene-functionalized ruthenium nanoparticles

    Science.gov (United States)

    Chen, Wei; Pradhan, Sulolit; Chen, Shaowei

    2011-05-01

    studies indicate that anthracene functionalization may be exploited as an effective route towards the manipulation of nanoparticle optoelectronic properties.Carbene-stabilized ruthenium nanoparticles were functionalized with anthryl moieties by olefin metathesis reactions with 9-vinylanthracene, at a surface concentration of about 19.7%, as estimated by 1H NMR spectroscopic measurements. Because of the conjugated metal-ligand interfacial bonding interactions, UV-vis measurements of the resulting nanoparticles showed a new broad absorption band centered at 612 nm, in addition to the peaks observed with monomeric vinylanthracene. FTIR measurements depicted apparent red-shifts of the aromatic vibrational stretches as compared to those of the monomeric vinylanthracene, suggestive of decreasing bonding order of the aromatic moieties as a result of extended conjugation between the particle-bound anthracene groups. Photoluminescence measurements confirmed the notion that effective intraparticle charge delocalization occurred by virtue of the conjugated metal-ligand interfacial bonding interactions, with apparent red-shifts of the excitation peaks and blue-shifts of the emission features, as compared to those of the monomeric vinylanthracene. The diminishment of the Stokes shift was, at least in part, attributed to the different chemical environments surrounding the anthryl moieties on the nanoparticle surface. Electronic conductivity measurements showed that because of the conjugated Ru z.dbd C π bonds, the activation energy for interparticle charge transport was about one order of magnitude lower than that observed with particles passivated by alkanethiolates. Additionally, whereas the original carbene-stabilized nanoparticles exhibited a semiconductor-metal transition within the temperature range of 100 to 320 K, anthracene-functionalized nanoparticles displayed apparent semiconducting behaviors with the ensemble conductivity increasing monotonically with temperature, most

  20. Investigation of plasma induced electrical and chemical factors and their contribution processes to plasma gene transfection.

    Science.gov (United States)

    Jinno, Masafumi; Ikeda, Yoshihisa; Motomura, Hideki; Kido, Yugo; Satoh, Susumu

    2016-09-01

    This study has been done to know what kind of factors in plasmas and processes on cells induce plasma gene transfection. We evaluated the contribution weight of three groups of the effects and processes, i.e. electrical, chemical and biochemical ones, inducing gene transfection. First, the laser produced plasma (LPP) was employed to estimate the contribution of the chemical factors. Second, liposomes were fabricated and employed to evaluate the effects of plasma irradiation on membrane under the condition without biochemical reaction. Third, the clathrin-dependent endocytosis, one of the biochemical processes was suppressed. It becomes clear that chemical factors (radicals and reactive oxygen/nitrogen species) do not work by itself alone and electrical factors (electrical current, charge and field) are essential to plasma gene transfection. It turned out the clathrin-dependent endocytosis is the process of the transfection against the 60% in all the transfected cells. The endocytosis and electrical poration are dominant in plasma gene transfection, and neither permeation through ion channels nor chemical poration is dominant processes. The simultaneous achievement of high transfection efficiency and high cell survivability is attributed to the optimization of the contribution weight among three groups of processes by controlling the weight of electrical and chemical factors. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Repeated Gene Transfection Impairs the Engraftment of Transplanted Porcine Neonatal Pancreatic Cells

    Directory of Open Access Journals (Sweden)

    Min Koo Seo

    2011-02-01

    Full Text Available BackgroundPreviously, we reported that neonatal porcine pancreatic cells transfected with hepatocyte growth factor (HGF gene in an Epstein-Barr virus (EBV-based plasmid (pEBVHGF showed improved proliferation and differentiation compared to those of the control. In this study, we examined if pancreatic cells transfected repeatedly with pEBVHGF can be successfully grafted to control blood glucose in a diabetes mouse model.MethodsNeonatal porcine pancreatic cells were cultured as a monolayer and were transfected with pEBVHGF every other day for a total of three transfections. The transfected pancreatic cells were re-aggregated and transplanted into kidney capsules of diabetic nude mice or normal nude mice. Blood glucose level and body weight were measured every other day after transplantation. The engraftment of the transplanted cells and differentiation into beta cells were assessed using immunohistochemistry.ResultsRe-aggregation of the pancreatic cells before transplantation improved engraftment of the cells and facilitated neovascularization of the graft. Right before transplantation, pancreatic cells that were transfected with pEBVHGF and then re-aggregated showed ductal cell marker expression. However, ductal cells disappeared and the cells underwent fibrosis in a diabetes mouse model two to five weeks after transplantation; these mice also did not show controlled blood glucose levels. Furthermore, pancreatic cells transplanted into nude mice with normal blood glucose showed poor graft survival regardless of the type of transfected plasmid (pCEP4, pHGF, or pEBVHGF.ConclusionFor clinical application of transfected neonatal porcine pancreatic cells, further studies are required to develop methods of overcoming the damage for the cells caused by repeated transfection and to re-aggregate them into islet-like structures.

  2. Use of SANS and SAXS in study of nanoparticles

    International Nuclear Information System (INIS)

    Goyal, P.S.; Aswal, V.K.

    2004-01-01

    Small Angle Neutron Scattering (SANS) and Small Angle X-ray Scattering (SAXS) are essentially diffraction experiments involving small (∼1 deg) scattering angles. Unlike conventional diffraction experiments (XRD or Neutron Diffraction) where one examines the specimen with an atomic resolution (∼0.2nm), SANS / SAXS are used to investigate structures on a length scale of ∼10 nm. Hence, SANS / SAXS are ideal techniques for studying the sizes and shapes of nano-particles. Because of differences in contrast for neutrons and x-rays, a combined SANS and SAXS study, at times, provides information about the core and the shell structure of nano particles. This talk will give an introduction to the techniques of SANS and SAXS and their applications in material science. Recent results of study of core-shell structure of micelles (nanoparticles of organic material) will be shown. (author)

  3. Demonstration by transfection studies that mutations in the adrenocorticotropin receptor gene are one cause of the hereditary syndrome of glucocorticoid deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Naville, D.; Barjhoux, L.; Jaillard, C. [Hopital Debrousse, Tours (France)] [and others

    1996-04-01

    The hereditary syndrome of unresponsiveness to ACTH is a rare autosomal recessive disorder characterized by low levels of serum cortisol and high levels of plasma ACTH. There is no cortisol response to exogenous ACTH. Recent cloning of the human ACTH receptor gene has enabled us to study this gene in patients with glucocorticoid deficiency. By using the PCR to amplify the coding sequence of the ACTH receptor gene, we identified three mutations in two unrelated patients. One mutation present in homozygous form converted the negatively charged Asp{sup 107}, located in the third transmembrane domain, to an uncharged Asn residue. The second patient was a compound heterozygote: the paternal allele contained a one-nucleotide insertion leading to a stop codon within the third extracellular loop, and the maternal allele contained a point mutation converting Cys{sup 235} to Phe, also in the third extracellular loop. Normal and mutant ACTH receptor genes were expressed in the M3 cell line, and intracellular cAMP production in response to ACTH was measured. For the mutant receptors, no response to physiological ACTH concentrations was detected, suggesting an impaired binding of ACTH to the receptors and/or an altered coupling to the adenylate cyclase effector. 24 refs., 6 figs., 2 tabs.

  4. Toward Contactless Biology: Acoustophoretic DNA Transfection

    Science.gov (United States)

    Vasileiou, Thomas; Foresti, Daniele; Bayram, Adem; Poulikakos, Dimos; Ferrari, Aldo

    2016-02-01

    Acoustophoresis revolutionized the field of container-less manipulation of liquids and solids by enabling mixing procedures which avoid contamination and loss of reagents due to the contact with the support. While its applications to chemistry and engineering are straightforward, additional developments are needed to obtain reliable biological protocols in a contactless environment. Here, we provide a first, fundamental step towards biological reactions in air by demonstrating the acoustophoretic DNA transfection of mammalian cells. We developed an original acoustophoretic design capable of levitating, moving and mixing biological suspensions of living mammalians cells and of DNA plasmids. The precise and sequential delivery of the mixed solutions into tissue culture plates is actuated by a novel mechanism based on the controlled actuation of the acoustophoretic force. The viability of the contactless procedure is tested using a cellular model sensitive to small perturbation of neuronal differentiation pathways. Additionally, the efficiency of the transfection procedure is compared to standard, container-based methods for both single and double DNA transfection and for different cell types including adherent growing HeLa cancer cells, and low adhesion neuron-like PC12 cells. In all, this work provides a proof of principle which paves the way to the development of high-throughput acoustophoretic biological reactors.

  5. Photo-transfection of mammalian cells via femtosecond laser pulses

    CSIR Research Space (South Africa)

    Mthunzi, P

    2009-06-01

    Full Text Available ). Transfection efficiencies between 40 - 63 % are recorded. We show for the first time that, due to their different sensitivity, surface receptors and membrane structure the cell lines mentioned above displayed varying photo-transfection efficiencies at different...

  6. Solid lipid nanoparticles with and without hydroxypropyl-β-cyclodextrin: a comparative study of nanoparticles designed for colonic drug delivery

    Science.gov (United States)

    Spada, Gianpiera; Gavini, Elisabetta; Cossu, Massimo; Rassu, Giovanna; Giunchedi, Paolo

    2012-03-01

    New solid lipid nanoparticles (SLN), composed of Compritol ATO888 (C) and hydroxypropyl-β-cyclodextrin (HP), were developed in order to study a new colon-specific formulation for diclofenac sodium (D) delivery. The prepared batches differ from each other by the molecular ratio between HP and D and by the composition of the matrix. Nanoparticles composed of an exclusively lipid matrix and nanoparticles with an oligomeric and lipid matrix were compared in order to establish the effect of both components on the drug delivery tests performed. The SLN preparation method was based on the oil/water hot homogenization process. Emulsions produced were cooled at room temperature and lyophilized in order to obtain dried nanoparticles; possible damage to nanoparticle shape and size was avoided by the addition of cryoprotectants to the aqueous dispersion of nanoparticles before exsiccation. An in vitro toxicity study was performed using CaCo2 cells to establish the safety of the prepared SLN. Data obtained showed that production method studied guarantees emulsions composed of nanosized drops which can be dried by lyophilization into SLN with a size range of 300-600 nm. In vitro and ex vivo tests demonstrated that dried SLN can be considered as colon delivery systems; however, the matrix composition as well as the presence of cryoprotectant on their surface influences the release and permeation rate of D. The in vitro toxicity studies indicated that the SLN are well tolerated.

  7. Solid lipid nanoparticles with and without hydroxypropyl-β-cyclodextrin: a comparative study of nanoparticles designed for colonic drug delivery

    International Nuclear Information System (INIS)

    Spada, Gianpiera; Gavini, Elisabetta; Cossu, Massimo; Rassu, Giovanna; Giunchedi, Paolo

    2012-01-01

    New solid lipid nanoparticles (SLN), composed of Compritol ATO888 (C) and hydroxypropyl-β-cyclodextrin (HP), were developed in order to study a new colon-specific formulation for diclofenac sodium (D) delivery. The prepared batches differ from each other by the molecular ratio between HP and D and by the composition of the matrix. Nanoparticles composed of an exclusively lipid matrix and nanoparticles with an oligomeric and lipid matrix were compared in order to establish the effect of both components on the drug delivery tests performed. The SLN preparation method was based on the oil/water hot homogenization process. Emulsions produced were cooled at room temperature and lyophilized in order to obtain dried nanoparticles; possible damage to nanoparticle shape and size was avoided by the addition of cryoprotectants to the aqueous dispersion of nanoparticles before exsiccation. An in vitro toxicity study was performed using CaCo 2 cells to establish the safety of the prepared SLN. Data obtained showed that production method studied guarantees emulsions composed of nanosized drops which can be dried by lyophilization into SLN with a size range of 300–600 nm. In vitro and ex vivo tests demonstrated that dried SLN can be considered as colon delivery systems; however, the matrix composition as well as the presence of cryoprotectant on their surface influences the release and permeation rate of D. The in vitro toxicity studies indicated that the SLN are well tolerated. (paper)

  8. Transfection efficiency of normal and cancer cell lines and monitoring of promoter activity by single-cell bioluminescence imaging.

    Science.gov (United States)

    Horibe, Tomohisa; Torisawa, Aya; Akiyoshi, Ryutaro; Hatta-Ohashi, Yoko; Suzuki, Hirobumi; Kawakami, Koji

    2014-02-01

    The bioluminescence system (luciferase reporter assay system) is widely used to study gene expression, signal transduction and other cellular activities. Although transfection of reporter plasmid DNA to mammalian cell lines is an indispensable experimental step, the transfection efficiency of DNA varies among cell lines, and several cell lines are not suitable for this type of assay because of the low transfection efficiency. In this study, we confirm the transfection efficiency of reporter DNA to several cancer and normal cell lines after transient transfection by single-cell imaging. Luminescence images could be obtained from living single cells after transient transfection, and the calculated transfection efficiency of this method was similar to that of the conventional reporter assay using a luminometer. We attempted to measure the activity of the Bip promoter under endoplasmic reticulum stress conditions using both high and low transfection efficiency cells for plasmid DNA at the single-cell level, and observed activation of this promoter even in cells with the lowest transfection efficiency. These results show that bioluminescence imaging of single cells is a powerful tool for the analysis of gene expression based on a reporter assay using limited samples such as clinical specimens or cells from primary culture, and could provide additional information compared with the conventional assay. Copyright © 2013 John Wiley & Sons, Ltd.

  9. Optimizing conditions for calcium phosphate mediated transient transfection

    Directory of Open Access Journals (Sweden)

    Ling Guo

    2017-03-01

    Conclusions: Calcium phosphate mediated transfection is the most low-cost approach to introduce recombinant DNA into culture cells. However, the utility of this procedure is limited in highly-differentiated cells. Here we describe the specific HBS-buffered saline, PH, glycerol shock, vortex strength, transfection medium, and particle concentrations conditions necessary to optimize this transfection method in highly differentiated cells.

  10. Chitosan-Graft-Polyethylenimine/DNA Nanoparticles as Novel Non-Viral Gene Delivery Vectors Targeting Osteoarthritis

    Science.gov (United States)

    Lv, Lulu; Zhao, Huiqing

    2014-01-01

    The development of safe and efficient gene carriers is the key to the clinical success of gene therapy. The present study was designed to develop and evaluate the chitosan-graft-polyethylenimine (CP)/DNA nanoparticles as novel non-viral gene vectors for gene therapy of osteoarthritis. The CP/DNA nanoparticles were produced through a complex coacervation of the cationic polymers with pEGFP after grafting chitosan (CS) with a low molecular weight (Mw) PEI (Mw = 1.8 kDa). Particle size and zeta potential were related to the weight ratio of CP:DNA, where decreases in nanoparticle size and increases in surface charge were observed as CP content increased. The buffering capacity of CP was significantly greater than that of CS. The transfection efficiency of CP/DNA nanoparticles was similar with that of the Lipofectamine™ 2000, and significantly higher than that of CS/DNA and PEI (25 kDa)/DNA nanoparticles. The transfection efficiency of the CP/DNA nanoparticles was dependent on the weight ratio of CP:DNA (w/w). The average cell viability after the treatment with CP/DNA nanoparticles was over 90% in both chondrocytes and synoviocytes, which was much higher than that of PEI (25 kDa)/DNA nanoparticles. The CP copolymers efficiently carried the pDNA inside chondrocytes and synoviocytes, and the pDNA was detected entering into nucleus. These results suggest that CP/DNA nanoparticles with improved transfection efficiency and low cytotoxicity might be a safe and efficient non-viral vector for gene delivery to both chondrocytes and synoviocytes. PMID:24392152

  11. A study on the effect of chemically synthesized magnetite nanoparticles on earthworm: Eudrilus eugeniae

    Science.gov (United States)

    Samrot, Antony V.; Justin, C.; Padmanaban, S.; Burman, Ujjala

    2017-02-01

    Most look into the benefits of the nanoparticles, but keeping aside the benefits; this study focuses on the impacts of nanoparticles on living systems. Improper disposal of nanoparticles into the environment is a subject of pollution or nano-pollution which in turn affects the flora and fauna in the ecosystem, particularly soil ecosystem. Thus, this study was done to understand the impacts of chemically synthesized magnetite nanoparticles on earthworm— Eudrilus eugeniae, a soil-dependent organism which acquires food and nutrition from decaying matters. The chemically synthesized magnetite nanoparticles were characterized by UV-visible spectrophotometry, Fourier transform infrared spectroscopy and field emission scanning electron microscopy. Earthworms were allowed to interact with different concentrations of synthesized nanoparticles and the effect of the nanoparticles was analysed by studying the phenotypic changes followed by histology and inductively coupled plasma optical emission spectrometry analyses.

  12. Stability study of sodium colistimethate-loaded lipid nanoparticles.

    Science.gov (United States)

    Moreno-Sastre, M; Pastor, M; Esquisabel, A; Sans, E; Viñas, M; Bachiller, D; Pedraz, J L

    2016-11-01

    In the last decades, the encapsulation of antibiotics into nanoparticulate carriers has gained increasing attention for the treatment of infectious diseases. Sodium colistimethate-loaded solid lipid nanoparticles (Colist-SLNs) and nanostructured lipid carriers (Colist-NLCs) were designed aiming to treat the pulmonary infection associated to cystic fibrosis patients. The nanoparticles were freeze-dried using trehalose as cryoprotectant. The stability of both nanoparticles was analysed over one year according to the International Conference of Harmonisation (ICH) guidelines by determining the minimum inhibitory concentration (MIC) against clinically isolated Pseudomonas aeruginosa strains and by studying their physico-chemical characteristics. The results showed that Colist-SLNs lost their antimicrobial activity at the third month; on the contrary, the antibacterial activity of Colist-NLCs was maintained throughout the study within an adequate range (MIC ≤16 μg/mL). In addition, Colist-NLCs exhibited suitable physico-chemical properties at 5 °C and 25 °C/60% relative humidity over one year. Altogether, Colist-NLCs proved to have better stability than Colist-SLNs.

  13. A study of atomic interaction between suspended nanoparticles and sodium atoms in liquid sodium

    International Nuclear Information System (INIS)

    Saito, Jun-ichi; Ara, Kuniaki

    2010-01-01

    A feasibility study of suppression of the chemical reactivity of sodium itself using an atomic interaction between nanoparticles and sodium atoms has been carried out. We expected that the atomic interaction strengthens when the nanoparticle metal is the transition element which has a major difference in electronegativity from sodium. We also calculated the atomic interaction between nanoparticle and sodium atoms. It became clear that the atomic bond between the nanoparticle atom and the sodium atom is larger than that between sodium atoms, and the charge transfer takes place to the nanoparticle atom from the sodium atom. Using sodium with suspended nanoparticles, the fundamental physical properties related to the atomic interaction were investigated to verify the atomic bond. The surface tension of sodium with suspended nanoparticles increased, and the evaporation rate of sodium with suspended nanoparticles also decreased compared with that of sodium. Therefore the presence of the atomic interaction between nanoparticles and sodium was verified from these experiments. Because the fundamental physical property changes by the atomic interaction, we expected changes in the chemical reactivity characteristics. The chemical reaction properties of sodium with suspended nanoparticles with water were investigated experimentally. The released reaction heat and the reaction rate of sodium with suspended nanoparticles were reduced than those of sodium. The influence of the charge state of nanoparticle on the chemical process with water was theoretically investigated to speculate on the cause of reaction suppression. The potential energy in both primary and side reactions changed by the charge transfer, and the free energy of activation of the reaction with water increased. Accordingly, the reaction barrier also increased. This suggests there is a possibility of the reduction in the reaction of sodium by the suspension of nanoparticles. Consequently the possibility of the

  14. Off-target responses in the HeLa proteome subsequent to transient plasmid-mediated transfection.

    Science.gov (United States)

    Hagen, Lars; Sharma, Animesh; Aas, Per Arne; Slupphaug, Geir

    2015-01-01

    Transient transfection of mammalian cells with plasmid expression vectors and chemical transfection reagents is widely used to study protein transport and dynamics as well as phenotypic alterations mediated by the overexpressed protein. Despite the undisputed impact of this technique, surprisingly little is known about the cellular effects mediated by the transfection process per se. Conceivably, off-target effects could have implications upon proteins or processes being studied and understanding the molecular pathways affected would add value to the interpretation of experimental observations subsequent to cell transfection. Here we have used a SILAC-based proteomic approach to study differentially expressed proteins after transfection of HeLa cells with ECFP vector using a commonly employed non-liposome based transfection reagent, Fugene®HD. Whereas the transfection reagent itself mediated minimal effects upon protein expression, 11 proteins were found to be significantly upregulated after transfection, all of which were associated with an interferon type I/II response. The upregulated proteins might potentially inflict major cellular processes such as RNA splicing, chromatin remodeling, post-translational protein modification and cell cycle control. The results were validated by western analysis as well as quantitative RT-PCR and this demonstrated that an essentially identical response was induced in HeLa by transfection using an empty pUC18 vector, which does not contain a mammalian virus promoter, as well as a liposome-based transfection reagent, Lipofectamine(TM)2000. Notably, no induction of the interferon response was observed in HEK293 cells, suggesting that these cells might be preferable to HeLa to avoid undesired off-target effects in transfection studies encompassing interferon-signaling and antiviral responses. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Novel mechanism of gene transfection by low-energy shock wave

    Science.gov (United States)

    Hoon Ha, Chang; Cheol Lee, Seok; Kim, Sunghyen; Chung, Jihwa; Bae, Hasuk; Kwon, Kihwan

    2015-01-01

    Extracorporeal shock wave (SW) therapy has been studied in the transfection of naked nucleic acids into various cell lines through the process of sonoporation, a process that affects the permeation of cell membranes, which can be an effect of cavitation. In this study, siRNAs were efficiently transfected into primary cultured cells and mouse tumor tissue via SW treatment. Furthermore SW-induced siRNA transfection was not mediated by SW-induced sonoporation, but by microparticles (MPs) secreted from the cells. Interestingly, the transfection effect of the siRNAs was transferable through the secreted MPs from human umbilical vein endothelial cell (HUVEC) culture medium after treatment with SW, into HUVECs in another culture plate without SW treatment. In this study, we suggest for the first time a mechanism of gene transfection induced by low-energy SW through secreted MPs, and show that it is an efficient physical gene transfection method in vitro and represents a safe therapeutic strategy for site-specific gene delivery in vivo. PMID:26243452

  16. Hydrophobic modification of polyethyleneimine for gene transfectants

    International Nuclear Information System (INIS)

    Kim, Sung Tae; Choi, Joon Sig; Jang, Hyung Suk; Suh, Hea Ran; Park, Jong Sang

    2001-01-01

    A new gene transfer system was developed by using polylipoplexes, which were prepared by hydrophobic modification of polyethyleneimine (PEI, MW 2000). PEI 25kDa is well known for its excellent transfection efficiency but it has extreme cytotoxicity; therefore, its application for medical use is strictly limited. PEI 2kDa is able to form complexes with DNA and has low cytotoxicity. However, unfortunately, it shows no transfection efficiency so it can not be a candidate carrier for gene therapy. We designed novel polycationic amphilphiles by conjugating hydrophobic moieties, such as cholesterol and myristate, to PEI 2kDa. Cholesterol-conjugated PEI (PEI-Chol: P10C, P17C and P30C) and myristate-conjugated PEI (PEI-Myr:P10M, P16M and P26M) are different from the other cationic lipids in that they can form lipopolyplexes with plasmid DNA that have extra multi-positive charges in their hydrophilic parts. From a different point of view, they are also considered to be PEI derivatives with a small proportion of hydrophobic moiety. As a result of the modification, PEI-Chol and PEI-Myr showed much enhanced transfection activity but somewhat increased cytotoxicity. We also examined the effect of the amount of hydrophobic moiety on lipopolyplex-mediated gene transfer and observed that P17C and P26M are the most effective carriers in the series of two groups. MTT assay indicated that the more myristyl groups were attached to PEI, the more injurious results were observed. In the case of PEI-Chol, however, the opposite tendency was observed

  17. Hematopoietic stem cell (CD34+) uptake of superparamagnetic iron oxide is enhanced by but not dependent on a transfection agent.

    Science.gov (United States)

    England, Timothy J; Bath, Philip M W; Abaei, Maryam; Auer, Dorothee; Jones, D Rhodri E

    2013-03-01

    Tracking the fate of cells after infusion would be a valuable asset for many stem cell therapies, but very few (cell) labels are approved for human therapeutic use. Superparamagnetic iron oxide particles (SPIO) can be internalized into stem cells in vitro to allow real-time tracking with gradient echo magnetic resonance imaging, but SPIO are approved for (diagnostic) imaging and not for (therapeutic) cell labeling in vivo. In this study, we investigated the possibility of labeling stem cells with an SPIO approved for patient use, albeit in a novel manner by enhancing uptake with the use of a transfection agent, also approved for patient use. Although there are many reports of hematopoietic stem cells being labeled with SPIO, there is some controversy regarding the efficiency of this and whether undifferentiated CD34+ progenitor (stem) cells are able to take up iron in the absence of a transfection agent to enhance the process. Human CD34+ cells were treated in vitro as follows: incubation with (i) medium only (control), (ii) ferumoxide (Endorem) and (iii) ferumoxide (Endorem) plus exposure to a transfection agent (protamine sulfate). Cells were incubated for 2, 4 and 24 hours and assessed for viability, differentiation capacity and visualized in vitro with 3-T magnetic resonance imaging. The cells were also analyzed by means of flow cytometry and morphology examined by electron microscopy. CD34+ hematopoietic progenitor cells can internalize ferumoxide (Endorem) independently of a transfection agent. However, uptake of ferumoxide is enhanced after exposure to protamine sulfate. Iron labeling of CD34+ cells in this manner does not affect cell viability and does not appear to affect the potential of the cells to grow in culture. Iron-labeled CD34+ cells can be visualized in vitro on 3-T magnetic resonance image scanning. Endorem and protamine sulfate can be combined to promote iron oxide nanoparticle uptake by CD34+ cells, and this methodology can potentially be used

  18. Modifying the chemistry of graphene with substrate selection: A study of gold nanoparticle formation

    Science.gov (United States)

    Zaniewski, Anna M.; Trimble, Christie J.; Nemanich, Robert J.

    2015-03-01

    Graphene and metal nanoparticle composites are a promising class of materials with unique electronic, optical, and chemical properties. In this work, graphene is used as a reducing surface to grow gold nanoparticles out of solution-based metal precursors. The nanoparticle formation is found to strongly depend upon the graphene substrate selection. The studied substrates include diamond, p-type silicon, aluminum oxide, lithium niobate, and copper. Our results indicate that the chemical properties of graphene depend upon this selection. For example, for the same reaction times and concentration, the reduction of gold chloride to gold nanoparticles on graphene/lithium niobate results in 3% nanoparticle coverage compared to 20% coverage on graphene/silicon and 60% on graphene/copper. On insulators, nanoparticles preferentially form on folds and edges. Energy dispersive X-ray analysis is used to confirm the nanoparticle elemental makeup.

  19. Modifying the chemistry of graphene with substrate selection: A study of gold nanoparticle formation

    International Nuclear Information System (INIS)

    Zaniewski, Anna M.; Trimble, Christie J.; Nemanich, Robert J.

    2015-01-01

    Graphene and metal nanoparticle composites are a promising class of materials with unique electronic, optical, and chemical properties. In this work, graphene is used as a reducing surface to grow gold nanoparticles out of solution-based metal precursors. The nanoparticle formation is found to strongly depend upon the graphene substrate selection. The studied substrates include diamond, p-type silicon, aluminum oxide, lithium niobate, and copper. Our results indicate that the chemical properties of graphene depend upon this selection. For example, for the same reaction times and concentration, the reduction of gold chloride to gold nanoparticles on graphene/lithium niobate results in 3% nanoparticle coverage compared to 20% coverage on graphene/silicon and 60% on graphene/copper. On insulators, nanoparticles preferentially form on folds and edges. Energy dispersive X-ray analysis is used to confirm the nanoparticle elemental makeup

  20. Studies on polyethylene glycol coating on NiFe2O4 nanoparticles for biomedical applications

    International Nuclear Information System (INIS)

    Phadatare, M.R.; Khot, V.M.; Salunkhe, A.B.; Thorat, N.D.; Pawar, S.H.

    2012-01-01

    The NiFe 2 O 4 nanoparticles were prepared by the combustion method and these nanoparticles were successfully coated with polyethylene glycol (PEG) for the possible biomedical applications such as magnetic resonance imaging, drug delivery, tissue repair, magnetic fluid hyperthermia etc. The structural and magnetic characterizations of NiFe 2 O 4 nanoparticles were carried out by x-ray diffraction and vibrating sample magnetometry techniques, respectively. The morphology of the uncoated and coated nanoparticles was studied by scanning electron microscopy. The existence of PEG layer on NiFe 2 O 4 nanoparticles was confirmed by fourier transform infrared spectroscopy technique. - Highlights: ► Synthesis of nanocrystalline NiFe 2 O 4 by the combustion method. ► Magnetic properties of the NiFe 2 O 4 nanoparticles at room temperature. ► Coating of NiFe 2 O 4 nanoparticles by Polyethylene glycol (PEG).

  1. Modifying the chemistry of graphene with substrate selection: A study of gold nanoparticle formation

    Energy Technology Data Exchange (ETDEWEB)

    Zaniewski, Anna M.; Trimble, Christie J.; Nemanich, Robert J. [Department of Physics, Arizona State University, Tempe, Arizona 85281 (United States)

    2015-03-23

    Graphene and metal nanoparticle composites are a promising class of materials with unique electronic, optical, and chemical properties. In this work, graphene is used as a reducing surface to grow gold nanoparticles out of solution-based metal precursors. The nanoparticle formation is found to strongly depend upon the graphene substrate selection. The studied substrates include diamond, p-type silicon, aluminum oxide, lithium niobate, and copper. Our results indicate that the chemical properties of graphene depend upon this selection. For example, for the same reaction times and concentration, the reduction of gold chloride to gold nanoparticles on graphene/lithium niobate results in 3% nanoparticle coverage compared to 20% coverage on graphene/silicon and 60% on graphene/copper. On insulators, nanoparticles preferentially form on folds and edges. Energy dispersive X-ray analysis is used to confirm the nanoparticle elemental makeup.

  2. Synthesis and Study of Silver Nanoparticles

    Science.gov (United States)

    Soloman, Sally D.; Bahadory, Mozghan; Jeyarajasingam, Aravindan V.; Rutkowsky, Susan A.; Boritz, Charles; Mulfinger, Lorraine

    2007-01-01

    A laboratory experiment was conducted in which the students synthesized yellow colloidal silver, estimate particle size using visible spectroscopy and studied aggregation effects. The students were thus introduced to nanotechnology along with other topics such as redox chemistry, limiting and excess reactants, spectroscopy and atomic size.

  3. Single-Tailed Lipidoids Enhance the Transfection Activity of Their Double-Tailed Counterparts.

    Science.gov (United States)

    Wu, Yihang; Li, Linxian; Chen, Qing; Su, Yi; Levkin, Pavel A; Davidson, Gary

    2016-01-11

    Cationic lipid-like molecules (lipidoids) are widely used for in vitro and in vivo gene delivery. Nearly all lipidoids developed to date employ double-tail or multiple-tail structures for transfection. Single-tail lipidoids are seldom considered for transfection as they have low efficiency in gene delivery. So far, there is no detailed study on the contribution to transfection efficiency of single-tail lipidoids when combined with standard double-tail lipidoids. Here, we use combinatorial chemistry to synthesize 17 double-tail and 17 single-tail lipidoids using thiol-yne and thiol-ene click chemistry, respectively. HEK 293T cells were used to analyze transfection efficiency by fluorescence microscopy and calculated based on the percentage of cells transfected. The size and zeta potential of liposomes and lipoplexes were characterized by dynamic light scattering (DLS). Intracellular DNA delivery and trafficking was further examined using confocal microscopy. Our study shows that combining single with double-tail lipidoids increases uptake of lipoplexes, as well as cellular transfection efficiency.

  4. Exploring the Correlation Between Lipid Packaging in Lipoplexes and Their Transfection Efficacy

    Science.gov (United States)

    Moghaddam, Behfar; McNeil, Sarah E.; Zheng, Qinguo; Mohammed, Afzal R.; Perrie, Yvonne

    2011-01-01

    Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent. PMID:24309311

  5. siRNA transfection in larvae of the barnacle Amphibalanus amphitrite

    KAUST Repository

    Zhang, G.

    2015-06-25

    RNA interference (RNAi) provides an efficient and specific technique for functional genomic studies. Yet, no successful application of RNAi has been reported in barnacles. In this study, siRNA against p38 MAPK was synthesized and then transfected into A. amphitrite larvae at either the nauplius or cyprid stage, or at both stages. Effects of siRNA transfection on the p38 MAPK level were hardly detectable in the cyprids when they were transfected at the nauplius stage. In contrast, larvae that were transfected at the cyprid stage showed lower levels of p38 MAPK than the blank and reagent controls. However, significantly decreased levels of phosphorylated p38 MAPK (pp38 MAPK) and reduced settlement rates were observed only in ‘double transfections’, in which larvae were exposed to siRNA solution at both the nauplius and cyprid stages. A relatively longer transfection time and more larval cells directly exposed to siRNA might explain the higher efficiency of double transfection experiments.

  6. Chitosan nanoparticles-trypsin interactions: Bio-physicochemical and molecular dynamics simulation studies.

    Science.gov (United States)

    Salar, Safoura; Mehrnejad, Faramarz; Sajedi, Reza H; Arough, Javad Mohammadnejad

    2017-10-01

    Herein, we investigated the effect of the chitosan nanoparticles (CsNP) on the structure, dynamics, and activity of trypsin. The enzyme activity in complex with the nanoparticles slightly increased, which represents the interactions between the nanoparticles and the enzyme. The kinetic parameters of the enzyme, K m and k cat , increased after adding the nanoparticles, resulting in a slight increase in the catalytic efficiency (k cat /K m ). However, the effect of the nanoparticles on the kinetic stability of trypsin has not exhibited significant variations. Fluorescence spectroscopy did not show remarkable changes in the trypsin conformation in the presence of the nanoparticles. The circular dichroism (CD) spectroscopy results also revealed the secondary structure of trypsin attached to the nanoparticles slightly changed. Furthermore, we used molecular dynamics (MD) simulation to find more information about the interaction mechanisms between the nanoparticles and trypsin. The root mean square deviation (RMSD) of Cα atoms results have shown that in the presence of the nanoparticles, trypsin was stable. The simulation and the calculation of the binding free energy demonstrate that the nonpolar interactions are the most important forces for the formation of stable nanoparticle-trypsin complex. This study has explicitly elucidated that the nanoparticles have not considerable effect on the trypsin. Copyright © 2017. Published by Elsevier B.V.

  7. Synthesis of Manganese Tetroxide Nanoparticles Using Precipitation and Study of Its Structure and Optical Characteristics

    Directory of Open Access Journals (Sweden)

    Reza Shokoohi

    2016-12-01

    Full Text Available Considering extensive applications of manganese tetroxide nanoparticles in various industries due to its special properties, conducting studies on how to achieve more suitable ways to produce smaller nanoparticles is of great importance. In this study, nanoparticles of manganese tetroxide (Mn3O4 were synthesized by a co-precipitation method. In order to determine the characteristics of the structure, size, and specific surface of the resulting nanoparticles, techniques such as XRD, BET, BJH, FESEM, and FTIR were employed. Also, the nanoparticles were quantified with EDS and their colony size was examined using DLS experiments. The findings revealed a production of crystalline manganese tetroxide nanoparticles with a space group of 141/amd (S.G. (141 and a molecular weight of 228.81 with the international code of ICSD Card # 89 - 4837. The specific surface area was 32.147 m2/g with a pore volume of 0.1041 cm3/g. The XRD and EDX analyses verify the production of the Mn3O4 nanoparticles. The size of the nanostructures is approximately 19 nm. The method used in this study could produce the Mn3O4 nanoparticles in a much easier way without the need for surfactants. Compared to the nanoparticles produced in other studies, the size of the nanoparticles produced in the present study is remarkably smaller. Moreover, less amount of the metal salt was used.

  8. Acidity-responsive gene delivery for "superfast" nuclear translocation and transfection with high efficiency.

    Science.gov (United States)

    Zhu, Jing-Yi; Zeng, Xuan; Qin, Si-Yong; Wan, Shuang-Shuang; Jia, Hui-Zhen; Zhuo, Ren-Xi; Feng, Jun; Zhang, Xian-Zheng

    2016-03-01

    In principle, not only efficient but rapid transfection is required since it can maximize the bioavailability of vector-carried gene prior to the cellular excretion. However, the "rapid" goal has been paid few attentions so far in the research field of vector-aided transfection. As a pioneering attempt, the present study designed a lysosome-targeting acidity-responsive nanoassembly as gene vectors, which proved the amazing potency to mediate the "Superfast" transnuclear gene transport and gene transfection with high efficiency in vitro and in vivo. The nanoassembly was constructed on the pH-reversible covalent boronic acid-diol coupling between 1,3-diol-rich oligoethylenimine (OEI-EHDO) and phenylboronic acid modified cholesterol (Chol-PBA). The rapid and efficient nuclei-tropic delivery and transfection was demonstrated to highly rely on the lysosome-acidity induced assembly destruction followed by the easy liberation of gene payloads inside cells. The nanoassembly-mediated transfection at 8 h can afford the outcome even comparable to that achieved at 48 h by the golden standard of PEI25k, and the transfection efficiency can still remain at a high level during 48 h. In contrast, time-dependent efficiency enhancement was identified for the transfections using PEI25k and OEI-EHDO as delivery vectors. Moreover, owing to the hydroxyl-rich surface, this delivery nanosystem presented strong tolerance to the serum-induced transfection inhibition that frequently occurred for the polycationic gene vectors such as PEI25k. The in vitro and in vivo results manifested the low toxicity of this bio-decomposable nanoassembly. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. SANS study to probe nanoparticle dispersion in nanocomposite membranes of aromatic polyamide and functionalized silica nanoparticles.

    Science.gov (United States)

    Jadav, Ghanshyam L; Aswal, Vinod K; Singh, Puyam S

    2010-11-01

    Silica nanoparticles produced from organically functionalized silicon alkoxide precursors were incorporated into polyamide film to produce a silica-polyamide nanocomposite membrane with enhanced properties. The dispersion of the silica nanoparticles in the nanocomposite membrane was characterized by performing small-angle neutron scattering (SANS) measurements on dilute reactant systems and dilute solution suspensions of the final product. Clear scattering of monodisperse spherical particles of 10-18 A R(g) were observed from dilute solutions of the initial reactant system. These silica nanoparticles initially reacted with diamine monomers of polyamide and subsequently were transformed into polyamide-coated silica nanoparticles; finally nanoparticle aggregates of 27-45 A R(g) were formed. The nanoparticle dispersion of the membrane as the nanosized aggregates is in corroboration with ring- or chain-like assemblies of the nanoparticles dispersed in the bulk polyamide phase as observed by transmission electron microscopy. It is demonstrated that dispersions of silica nanoparticles as the nanosized aggregates in the polyamide phase could be achieved in the nanocomposite membrane with a silica content up to about 2 wt.%. Nanocomposite membranes with higher silica loading approximately 10 wt.% lead to the formation of large aggregates of sizes over 100 A R(g) in addition to the nanosized aggregates. Copyright 2010 Elsevier Inc. All rights reserved.

  10. Study of structural modification of PVA by incorporating Ag nanoparticles

    DEFF Research Database (Denmark)

    Saini, I.; Sharma, A.; Rozra, J.

    2016-01-01

    Nanocomposites of PVA with Ag nanoparticles dispersed in it were synthesized using solution casting method. The morphology and size distribution of Ag nanoparticles embedded in PVA matrix were obtained by transmission electron microscopy (TEM) and Field emission scanning electron microscopy (FE......-SEM). Raman spectroscopy was used to examine structural changes taking place inside polyvinyl alcohol (PVA) matrix due to incorporation of Ag nanoparticle. Raman analysis indicates that Ag nanoparticles interact with PVA through H-bonding. © 2016 Author(s)....

  11. Study of structural modification of PVA by incorporating Ag nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Saini, Isha; Sharma, Annu, E-mail: asharma@kuk.ac.in; Rozra, Jyoti; Aggarwal, Sanjeev [Department of Physics, Kurukshetra University, Kurukshetra 136119 (India); Dhiman, Rajnish [Department of Physics and Chemistry, University of Southern Denmark, Campusvej 55, DK-5230 Odense M (Denmark); Sharma, Pawan K. [Department of Chemistry, Kurukshetra University, Kurukshetra 136119 (India)

    2016-05-23

    Nanocomposites of PVA with Ag nanoparticles dispersed in it were synthesized using solution casting method. The morphology and size distribution of Ag nanoparticles embedded in PVA matrix were obtained by transmission electron microscopy (TEM) and Field emission scanning electron microscopy (FE-SEM). Raman spectroscopy was used to examine structural changes taking place inside polyvinyl alcohol (PVA) matrix due to incorporation of Ag nanoparticle. Raman analysis indicates that Ag nanoparticles interact with PVA through H-bonding.

  12. Transfection of rat embryo cells with mutant p53 increases the intrinsic radiation resistance

    International Nuclear Information System (INIS)

    Pardo, F.S.; Su, M.; Gerweck, L.; Schmidt, E.V.; Borek, C.; Preffer, F.; Dombkowski, D.

    1994-01-01

    Dominant oncogenic sequences have been shown to modulate the intrinsic radiation sensitivity of cells of both human and murine tumor cell lines. Whether transfection with candidate tumor-suppressor genes can modulate intrinsic radiation sensitivity is unknown. The data presented here demonstrate that transfection of rat embryo cells with a mutant p53 allele can increase the intrinsic radiation resistance of cells in vitro. First, transfection with mutant p53 resulted in transformed cellular morphology. Second, the transfected clone and the corresponding pooled population of transfected clones were more resistant to ionizing radiation in vitro. Last, analyses of the parameters of cell kinetics suggested that the radiobiological effects were unlikely to be due to altered parameters of cell kinetics at the time of irradiation, suggesting that mutant p53 altered the intrinsic radiation resistance of transfected cells by a more direct mechanism. Further experimentation will be necessary to develop a mechanistic approach for the study of these alterations. 29 refs., 3 figs., 2 tabs

  13. Persistent human cardiac Na+ currents in stably transfected mammalian cells

    Science.gov (United States)

    Wang, Ging Kuo; Russell, Gabriella; Wang, Sho-Ya

    2013-01-01

    Miniature persistent late Na+ currents in cardiomyocytes have been linked to arrhythmias and sudden death. The goals of this study are to establish a stable cell line expressing robust persistent cardiac Na+ currents and to test Class 1 antiarrhythmic drugs for selective action against resting and open states. After transient transfection of an inactivation-deficient human cardiac Na+ channel clone (hNav1.5-CW with L409C/A410W double mutations), transfected mammalian HEK293 cells were treated with 1 mg/ml G-418. Individual G-418-resistant colonies were isolated using glass cylinders. One colony with high expression of persistent Na+ currents was subjected to a second colony selection. Cells from this colony remained stable in expressing robust peak Na+ currents of 996 ± 173 pA/pF at +50 mV (n = 20). Persistent late Na+ currents in these cells were clearly visible during a 4-second depolarizing pulse albeit decayed slowly. This slow decay is likely due to slow inactivation of Na+ channels and could be largely eliminated by 5 μM batrachotoxin. Peak cardiac hNav1.5-CW Na+ currents were blocked by tetrodotoxin with an IC50 value of 2.27 ± 0.08 μM (n = 6). At clinic relevant concentrations, Class 1 antiarrhythmics are much more selective in blocking persistent late Na+ currents than their peak counterparts, with a selectivity ratio ranging from 80.6 (flecainide) to 3 (disopyramide). We conclude that (1) Class 1 antiarrhythmics differ widely in their resting- vs. open-channel selectivity, and (2) stably transfected HEK293 cells expressing large persistent hNav1.5-CW Na+ currents are suitable for studying as well as screening potent open-channel blockers. PMID:23695971

  14. Gold and gold-copper nanoparticles in 2-propanol: A radiation chemical study

    International Nuclear Information System (INIS)

    Dey, G.R.

    2011-01-01

    The studies on the reduction of Au 3+ to gold nanoparticles in presence and absence of Cu 2+ under deoxygenated conditions in 2-propanol by radiolytic method have been carried out. On γ-radiolysis, preliminary yellow colored solution of Au 3+ changed to purple color owing to gold nanoparticles formation, which exhibits an absorption peak at around 540 nm. In the presence of Cu 2+ , absorption of gold-copper nanoparticles, which was also produced during γ-radiolysis, was red shifted in contrast to the system containing no Cu 2+ . Under DLS studies the sizes of gold nanoparticles in the absence and the presence of Cu 2+ were found to be larger (>400 nm). However, in presence of polyethylene glycol, a stabilizer the nanoparticle sizes became smaller, sizes measured for gold and gold-copper nanoparticles are 40 and 140 nm, respectively. Moreover, the change in UV-vis spectra in the Cu 2+ and Au 3+ mixed system highlights the formation of gold-copper nanoparticles in core-shell type arrangement. - Highlights: → Present radiation chemical study highlights high reactivity of Au ·2+ with Cu 2+ . → Absorption of gold-copper nanoparticles is blue shifted as compared to copper nanoparticles. → Change in UV-vis spectra with dose emphasizes core-shell type arrangement of Au-Cu nanoparticles.

  15. Inner ear gene transfection in neonatal mice using adeno-associated viral vector: a comparison of two approaches.

    Directory of Open Access Journals (Sweden)

    Li Xia

    Full Text Available Local gene transfection is a promising technique for the prevention and/or correction of inner ear diseases, particularly those resulting from genetic defects. Adeno-associated virus (AAV is an ideal viral vector for inner ear gene transfection because of its safety, stability, long-lasting expression, and its high tropism for many different cell types. Recently, a new generation of AAV vectors with a tyrosine mutation (mut-AAV has demonstrated significant improvement in transfection efficiency. A method for inner ear gene transfection via the intact round window membrane (RWM has been developed in our laboratory. This method has not been tested in neonatal mice, an important species for the study of inherited hearing loss. Following a preliminary study to optimize the experimental protocol in order to reduce mortality, the present study investigated inner ear gene transfection in mice at postnatal day 7. We compared transfection efficiency, the safety of the scala tympani injection via RWM puncture, and the trans-RWM diffusion following partial digestion with an enzyme technique. The results revealed that approximately 47% of inner hair cells (IHCs and 17% of outer hair cells (OHCs were transfected via the trans-RWM approach. Transfection efficiency via RWM puncture (58% and 19% for IHCs and OHCs, respectively was slightly higher, but the difference was not significant.

  16. Biological Monitoring of Inhaled Nanoparticles in Patients: An Appealing Approach To Study Causal Link between Human Respiratory Pathology and Exposure to Nanoparticles.

    Science.gov (United States)

    Forest, Valérie; Vergnon, Jean-Michel; Pourchez, Jérémie

    2017-09-18

    Although necessary, in vitro and in vivo studies are not fully successful at predicting nanomaterials toxicity. We propose to associate such assays to the biological monitoring of nanoparticles in clinical samples to get more relevant data on the chemical and physical nature and dose of nanoparticles found in humans. The concept is to establish the load of nanoparticles in biological samples of patients. Then, by comparing samples from different patient groups, nanoparticles of interest could be identified and a potential link between a given nanoparticle type and toxicity could be suggested. It must be confirmed by investigating the biological effects induced by these nanoparticles using in vitro or in vivo models (mechanistic or dose-response studies). This translational approach from the bedside to the bench and vice versa could allow a better understanding of the nanoparticle effects and mechanisms of toxicity that can contribute, at least in part, to a disease.

  17. Graphene and carbon nanotube nanocomposite for gene transfection.

    Science.gov (United States)

    Hollanda, L M; Lobo, A O; Lancellotti, M; Berni, E; Corat, E J; Zanin, H

    2014-06-01

    Graphene and carbon nanotube nanocomposite (GCN) was synthesised and applied in gene transfection of pIRES plasmid conjugated with green fluorescent protein (GFP) in NIH-3T3 and NG97 cell lines. The tips of the multi-walled carbon nanotubes (MWCNTs) were exfoliated by oxygen plasma etching, which is also known to attach oxygen content groups on the MWCNT surfaces, changing their hydrophobicity. The nanocomposite was characterised by high resolution scanning electron microscopy; energy-dispersive X-ray, Fourier transform infrared and Raman spectroscopies, as well as zeta potential and particle size analyses using dynamic light scattering. BET adsorption isotherms showed the GCN to have an effective surface area of 38.5m(2)/g. The GCN and pIRES plasmid conjugated with the GFP gene, forming π-stacking when dispersed in water by magnetic stirring, resulting in a helical wrap. The measured zeta potential confirmed that the plasmid was connected to the nanocomposite. The NIH-3T3 and NG97 cell lines could phagocytize this wrap. The gene transfection was characterised by fluorescent protein produced in the cells and pictured by fluorescent microscopy. Before application, we studied GCN cell viability in NIH-3T3 and NG97 line cells using both MTT and Neutral Red uptake assays. Our results suggest that GCN has moderate stability behaviour as colloid solution and has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity and good transfection efficiency. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Synthesis and in vitro release study of ibuprofen-loaded gelatin graft copolymer nanoparticles.

    Science.gov (United States)

    Haroun, Ahmed A; El-Halawany, N R; Loira-Pastoriza, C; Maincent, P

    2014-01-01

    This work deals with the preparation, characterization and in vitro release study of IBU-loaded gel graft copolymer nanoparticles. Gelatin (Gel) graft copolymer nanoparticles were prepared using styrene (Sty) and/or 2-hydroxyethyl methacrylate (HEMA) monomers in the presence of potassium persulfate and glutaraldehyde as an initiator and cross-linker, respectively. The prepared nanoparticles as sustained release drug carriers were investigated using the nonsteriodal anti-inflammatory model drug, ibuprofen (IBU). The prepared nanoparticles as sustained release drug carriers were investigated using the nonsteriodal anti-inflammatory model drug, IBU. The prepared Gel/HEMA and Gel/Sty nanoparticles exhibited particles size ranging from 15 to 17 nm and from 0.42 to 5 mm, respectively. The dissolution of IBU in phosphate buffer, pH 7.4, at 37°C from the prepared nanoparticles was evaluated using UV spectroscopy. In addition, the prepared nanoparticles were characterized using Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), transmitting electron microscope (TEM) and zeta potential/particle size analyzer. In vitro dissolution study showed that the dissolution rates of the crosslinked nanoparticles were retarded relative to the uncrosslinked ones. Moreover, the released amount constantly decreases with increasing gluteraldehyde content in the gel nanoparticles. Crosslinked gel-based graft copolymers exhibited slow IBU release within six hours. Furthermore, results from different characterization techniques such as TEM, particles size and zeta potential measurements confirmed the formation of pH-responsive gel-graft copolymer nanoparticles.

  19. Comparative toxicity study of Ag, Au, and Ag-Au bimetallic nanoparticles on Daphnia magna.

    Science.gov (United States)

    Li, Ting; Albee, Brian; Alemayehu, Matti; Diaz, Rocio; Ingham, Leigha; Kamal, Shawn; Rodriguez, Maritza; Bishnoi, Sandra Whaley

    2010-09-01

    A comparative assessment of the 48-h acute toxicity of aqueous nanoparticles synthesized using the same methodology, including Au, Ag, and Ag-Au bimetallic nanoparticles, was conducted to determine their ecological effect in freshwater environments through the use of Daphnia magna, using their mortality as a toxicological endpoint. D. magna are one of the standard organisms used for ecotoxicity studies due to their sensitivity to chemical toxicants. Particle suspensions used in toxicity testing were well-characterized through a combination of absorbance measurements, atomic force or electron microscopy, flame atomic absorption spectrometry, and dynamic light scattering to determine composition, aggregation state, and particle size. The toxicity of all nanoparticles tested was found to be dose and composition dependent. The concentration of Au nanoparticles that killed 50% of the test organisms (LC(50)) ranged from 65-75 mg/L. In addition, three different sized Ag nanoparticles (diameters = 36, 52, and 66 nm) were studied to analyze the toxicological effects of particle size on D. magna; however, it was found that toxicity was not a function of size and ranged from 3-4 μg/L for all three sets of Ag nanoparticles tested. This was possibly due to the large degree of aggregation when these nanoparticles were suspended in standard synthetic freshwater. Moreover, the LC(50) values for Ag-Au bimetallic nanoparticles were found to be between that of Ag and Au but much closer to that of Ag. The bimetallic particles containing 80% Ag and 20% Au were found to have a significantly lower toxicity to Daphnia (LC(50) of 15 μg/L) compared to Ag nanoparticles, while the toxicity of the nanoparticles containing 20% Ag and 80% Au was greater than expected at 12 μg/L. The comparison results confirm that Ag nanoparticles were much more toxic than Au nanoparticles, and that the introduction of gold into silver nanoparticles may lower their environmental impact by lowering the amount

  20. Nanoparticles as conjugated delivery agents for therapeutic applications

    Science.gov (United States)

    Muroski, Megan Elizabeth

    This dissertation explores the use of nanoparticles as conjugated delivery agents. Chapter 1 is a general introduction. Chapter 2 discusses the delivery by a nanoparticle platform provides a method to manipulate gene activation, by taking advantage of the high surface area of a nanoparticle and the ability to selectively couple a desired biological moiety to the NP surface. The nanoparticle based transfection approach functions by controlled release of gene regulatory elements from a 6 nm AuNP (gold nanoparticle) surface. The endosomal release of the regulatory elements from the nanoparticle surface results in endogenous protein knockdown simultaneously with exogenous protein expression for the first 48 h. The use of fluorescent proteins as the endogenous and exogenous signals for protein expression enables the efficiency of co-delivery of siRNA (small interfering RNA) for GFP (green fluorescent protein) knockdown and a dsRed-express linearized plasmid for induction to be optically analyzed in CRL-2794, a human kidney cell line expressing an unstable green fluorescent protein. Delivery of the bimodal nanoparticle in cationic liposomes results in 20% GFP knockdown within 24 h of delivery and continues exhibiting knockdown for up to 48 h for the bimodal agent. Simultaneous dsRed expression is observed to initiate within the same time frame with expression levels reaching 34% after 25 days although cells have divided approximately 20 times, implying daughter cell transfection has occurred. Fluorescence cell sorting results in a stable colony, as demonstrated by Western blot analysis. The simultaneous delivery of siRNA and linearized plasmid DNA on the surface of a single nanocrystal provides a unique method for definitive genetic control within a single cell and leads to a very efficient cell transfection protocol. In Chapter 3, we wanted to understand the NP complex within the cell, and to look at the dynamics of release utilizing nanometal surface energy transfer as

  1. Study of Ag and Au Nanoparticles Synthesized by Arc Discharge in Deionized Water

    Directory of Open Access Journals (Sweden)

    Der-Chi Tien

    2010-01-01

    Full Text Available The paper presents a study of Ag and Au nanofluids synthesized by the arc discharge method (ADM in deionized water. The metallic Ag nanoparticle (Ag0 and ionic Ag (Ag+ have played an important role in the battle against germs which are becoming more drug-resistant every year. Our study indicates that Ag nanoparticle suspension (SNPS fabricated by using ADM without added surfactants exclusively contains the metallic Ag nanoparticle and ionic Ag. Besides that, the ADM in deionized water has also been employed for the fabrication process of Au nanoparticles. The experimental results indicate that the prepared Ag nanoparticles can react with the dissolved H2CO3 in deionized water, leading to the formation of Ag2CO3. Significantly different to Ag, the prepared Au nanoparticles with their surfaces bonded by oxygen are suspended in deionized water by the formation of hydrogen bonded with the neighboring water molecules.

  2. Quantification of Functionalised Gold Nanoparticle-Targeted Knockdown of Gene Expression in HeLa Cells

    Science.gov (United States)

    Jiwaji, Meesbah; Sandison, Mairi E.; Reboud, Julien; Stevenson, Ross; Daly, Rónán; Barkess, Gráinne; Faulds, Karen; Kolch, Walter; Graham, Duncan; Girolami, Mark A.; Cooper, Jonathan M.; Pitt, Andrew R.

    2014-01-01

    Introduction Gene therapy continues to grow as an important area of research, primarily because of its potential in the treatment of disease. One significant area where there is a need for better understanding is in improving the efficiency of oligonucleotide delivery to the cell and indeed, following delivery, the characterization of the effects on the cell. Methods In this report, we compare different transfection reagents as delivery vehicles for gold nanoparticles functionalized with DNA oligonucleotides, and quantify their relative transfection efficiencies. The inhibitory properties of small interfering RNA (siRNA), single-stranded RNA (ssRNA) and single-stranded DNA (ssDNA) sequences targeted to human metallothionein hMT-IIa are also quantified in HeLa cells. Techniques used in this study include fluorescence and confocal microscopy, qPCR and Western analysis. Findings We show that the use of transfection reagents does significantly increase nanoparticle transfection efficiencies. Furthermore, siRNA, ssRNA and ssDNA sequences all have comparable inhibitory properties to ssDNA sequences immobilized onto gold nanoparticles. We also show that functionalized gold nanoparticles can co-localize with autophagosomes and illustrate other factors that can affect data collection and interpretation when performing studies with functionalized nanoparticles. Conclusions The desired outcome for biological knockdown studies is the efficient reduction of a specific target; which we demonstrate by using ssDNA inhibitory sequences targeted to human metallothionein IIa gene transcripts that result in the knockdown of both the mRNA transcript and the target protein. PMID:24926959

  3. Implementing atomic force microscopy (AFM) for studying kinetics of gold nanoparticle's growth

    DEFF Research Database (Denmark)

    Georgiev, P.; Bojinova, A.; Kostova, B.

    2013-01-01

    In a novel experimental approach Atomic Force Microscopy (AFM) was applied as a tool for studying the kinetics of gold nanoparticle growth. The gold nanoparticles were obtained by classical Turkevich citrate synthesis at two different temperatures. From the analysis of AFM images during...... the synthesis process the nanoparticle s' sizes were obtained. To demonstrate the applicability and the reliability of the proposed experimental approach we studied the nanoparticles growth at two different temperatures by spectrophotometric measurements and compared them with the results from AFM experimental...

  4. Repeated Aurora-A siRNA Transfection Results in Effective Apoptosis of A549 Cells Compared to Single Transfection.

    Science.gov (United States)

    Wang, Zhonghua; Sun, Wenwu; Cao, Jianping; Cui, Haiyang; Ma, Zhuang

    2016-01-01

    Suppression of Aurora kinase A (Aurora-A, AURKA) by Aurora-A siRNA has been proposed for lung tumor treatment. However, protocols using single administration have shown little benefit in some types of lung tumor. Given that transfection efficiency of Aurora-A siRNA is low due to tightly packed cells in the tumor, we hypothesized that repeated administration would result in efficient cell apoptosis. We compared single vs. repeated transfection (thrice) in A549 cells by transfecting Aurora-A siRNA (siA) on the 1st or 1st, 2nd and 3rd day after cell seeding. A random sequence was used as the negative siRNA control (siC). Cells in the single transfection group received only transfection reagent without siRNAs on the 2nd and 3rd day. Two days after the third transfection, both single and repeated siA administration decreased mRNA expression of Aurora-A and cell viability compared to no administration and siC single administration. However, the decrease in these two indices with repeated transfection was more obvious than that following single administration: cell viability decreased to 72.8 ± 3.05% (p transfection and to 64.2 ± 1.99% (p transfection, compared with normal control cells, respectively. Gene expression decreased to 17 ± 16.6% (p transfection and to 43.2 ± 13.0% (p transfection. Compared to single transfection, repeated Aurora-A siRNA transfection decreased Aurora-A, which, in turn, resulted in effective apoptosis of A549 cells.

  5. Interaction studies between biosynthesized silver nanoparticle with calf thymus DNA and cytotoxicity of silver nanoparticles

    Science.gov (United States)

    Roy, Swarup; Sadhukhan, Ratan; Ghosh, Utpal; Das, Tapan Kumar

    2015-04-01

    The interaction of calf thymus DNA (CTDNA) with silver nanoparticles (SNP) has been investigated following spectroscopic studies, analysis of melting temperature (Tm) curves and hydrodynamic measurement. In spectrophotometric titration and thermal denaturation studies of CTDNA it was found that SNP can form a complex with double-helical DNA and the increasing value of Tm also supported the same. The association constant of SNP with DNA from UV-Vis study was found to be 4.1 × 103 L/mol. The fluorescence emission spectra of intercalated ethidium bromide (EB) with increasing concentration of SNP represented a significant reduction of EB intensity and quenching of EB fluorescence. The results of circular dichroism (CD) suggested that SNP can change the conformation of DNA. From spectroscopic, hydrodynamic, and DNA melting studies, SNP has been found to be a DNA groove binder possessing partial intercalating property. Cell cytotoxicity of SNP was compared with that of normal silver salt solution on HeLa cells. Our results show that SNP has less cytotoxicity compared to its normal salt solution and good cell staining property.

  6. Engineering of magnetic DNA nanoparticles for tumor-targeted therapy

    International Nuclear Information System (INIS)

    Hosseinkhani, Hossein; Chen Yiru; He Wenjie; Hong Poda; Yu, Dah-Shyong; Domb, Abraham J.

    2013-01-01

    This study aims to engineer novel targeted delivery system composed of magnetic DNA nanoparticles to be effective as an efficient targeted gene therapy vehicle for tumor therapy. A polysaccharide, dextran, was chosen as the vector of plasmid DNA-encoded NK4 that acts as an HGF-antagonist and anti-angiogenic regulator for inhibitions of tumor growth, invasion, and metastasis. Spermine (Sm) was chemically introduced to the hydroxyl groups of dextran to obtain dextran-Sm. When Fe 2+ solution was added to the mixture of dextran-Sm and a plasmid DNA, homogenous DNA nanoparticles were formed via chemical metal coordination bonding with average size of 230 nm. Characterization of DNA nanoparticles was performed via dynamic light scattering measurement, electrophoretic light scattering measurement, as well as transmission electron microscope. DNA nanoparticles effectively condensed plasmid DNA into nanoparticles and enhanced the stability of DNA, while significantly improved transfection efficiency in vitro and tumor accumulation in vivo. In addition, magnetic DNA nanoparticles exhibited high efficiency in antitumor therapy with regards to tumor growth as well as survival of animals evaluated in the presence of external magnetic field. We conclude that the magnetic properties of these DNA nanoparticles would enhance the tracking of non-viral gene delivery systems when administrated in vivo in a test model. These findings suggest that DNA nanoparticles effectively deliver DNA to tumor and thereby inhibiting tumor growth.

  7. Engineering of magnetic DNA nanoparticles for tumor-targeted therapy

    Energy Technology Data Exchange (ETDEWEB)

    Hosseinkhani, Hossein, E-mail: hosseinkhani@yahoo.com [Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology (Taiwan Tech) (China); Chen Yiru [National Yang-Ming University, Department of Biomedical Engineering (China); He Wenjie; Hong Poda [Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology (Taiwan Tech) (China); Yu, Dah-Shyong [Nanomedicine Research Center, National Defense Medical Center (China); Domb, Abraham J. [Institute of Drug Research, The Center for Nanoscience and Nanotechnology, School of Pharmacy, Faculty of Medicine, Hebrew University of Jerusalem (Israel)

    2013-01-15

    This study aims to engineer novel targeted delivery system composed of magnetic DNA nanoparticles to be effective as an efficient targeted gene therapy vehicle for tumor therapy. A polysaccharide, dextran, was chosen as the vector of plasmid DNA-encoded NK4 that acts as an HGF-antagonist and anti-angiogenic regulator for inhibitions of tumor growth, invasion, and metastasis. Spermine (Sm) was chemically introduced to the hydroxyl groups of dextran to obtain dextran-Sm. When Fe{sup 2+} solution was added to the mixture of dextran-Sm and a plasmid DNA, homogenous DNA nanoparticles were formed via chemical metal coordination bonding with average size of 230 nm. Characterization of DNA nanoparticles was performed via dynamic light scattering measurement, electrophoretic light scattering measurement, as well as transmission electron microscope. DNA nanoparticles effectively condensed plasmid DNA into nanoparticles and enhanced the stability of DNA, while significantly improved transfection efficiency in vitro and tumor accumulation in vivo. In addition, magnetic DNA nanoparticles exhibited high efficiency in antitumor therapy with regards to tumor growth as well as survival of animals evaluated in the presence of external magnetic field. We conclude that the magnetic properties of these DNA nanoparticles would enhance the tracking of non-viral gene delivery systems when administrated in vivo in a test model. These findings suggest that DNA nanoparticles effectively deliver DNA to tumor and thereby inhibiting tumor growth.

  8. Engineering of magnetic DNA nanoparticles for tumor-targeted therapy

    Science.gov (United States)

    Hosseinkhani, Hossein; Chen, Yi-Ru; He, Wenjie; Hong, Po-Da; Yu, Dah-Shyong; Domb, Abraham J.

    2013-01-01

    This study aims to engineer novel targeted delivery system composed of magnetic DNA nanoparticles to be effective as an efficient targeted gene therapy vehicle for tumor therapy. A polysaccharide, dextran, was chosen as the vector of plasmid DNA-encoded NK4 that acts as an HGF-antagonist and anti-angiogenic regulator for inhibitions of tumor growth, invasion, and metastasis. Spermine (Sm) was chemically introduced to the hydroxyl groups of dextran to obtain dextran-Sm. When Fe2+ solution was added to the mixture of dextran-Sm and a plasmid DNA, homogenous DNA nanoparticles were formed via chemical metal coordination bonding with average size of 230 nm. Characterization of DNA nanoparticles was performed via dynamic light scattering measurement, electrophoretic light scattering measurement, as well as transmission electron microscope. DNA nanoparticles effectively condensed plasmid DNA into nanoparticles and enhanced the stability of DNA, while significantly improved transfection efficiency in vitro and tumor accumulation in vivo. In addition, magnetic DNA nanoparticles exhibited high efficiency in antitumor therapy with regards to tumor growth as well as survival of animals evaluated in the presence of external magnetic field. We conclude that the magnetic properties of these DNA nanoparticles would enhance the tracking of non-viral gene delivery systems when administrated in vivo in a test model. These findings suggest that DNA nanoparticles effectively deliver DNA to tumor and thereby inhibiting tumor growth.

  9. Structural and photoluminescence studies of TiO{sub 2} nanoparticles synthesized by solution combustion method

    Energy Technology Data Exchange (ETDEWEB)

    Balamurugan, M., E-mail: chem.muruga@gmail.com; Silambarasan, M. [Centre for Photonics and Nanotechnology, Department of Science, Sona College of Technology, Salem – 636 005, Tamilnadu (India); Saravanan, S. [Centre for Photonics and Nanotechnology, Department of Science, Sona College of Technology, Salem – 636 005, Tamilnadu (India); Department of Frontier Materials, Nagoya Institute of Technology, Nagoya - 466-8555 (Japan); Soga, Tetsuo [Department of Frontier Materials, Nagoya Institute of Technology, Nagoya - 466-8555 (Japan)

    2015-06-24

    In this study titanium dioxide nanoparticle is prepared by simple solution combustion method. The powder X-ray diffraction pattern indicates the prepared titanium dioxide nanoparticles crystalline nature with tetragonal structure. Also it shows the nanoparticle is anatase and rutile mixed phase. The Field Emission Scanning Electron Microscopy image shows the nanostructure of particles in the size range about 50 nm. Room temperature photoluminescence shows intrinsic defects of oxygen vacancies.

  10. Coarsening of Pd nanoparticles in an oxidizing atmosphere studied by in situ TEM

    DEFF Research Database (Denmark)

    Simonsen, Søren Bredmose; Chorkendorff, Ib; Dahl, Søren

    2016-01-01

    The coarsening of supported palladium nanoparticles in an oxidizing atmosphere was studied in situ by means of transmission electron microscopy (TEM). Specifically, the Pd nanoparticles were dispersed on a planar and amorphous Al2O3 support and were observed during the exposure to 10 mbar technical...... for the Ostwald ripening process indicates that the observed change in the particle size distribution can be accounted for by wetting of the Al2O3 support by the larger Pd nanoparticles....

  11. Small angle neutron scattering study of disordered and crystalline iron nanoparticle assemblies

    International Nuclear Information System (INIS)

    Farrell, D.F.; Ijiri, Y.; Kelly, C.V.; Borchers, J.A.; Rhyne, J.J.; Ding, Y.; Majetich, S.A.

    2006-01-01

    Monodisperse surfactant-coated iron nanoparticles are used to form both disordered nanoparticle assemblies and ordered face-centered cubic nanoparticle crystals. The structural order is probed by small angle X-ray scattering, and the magnetic scattering is studied using small angle neutron scattering. The magnetic scattering corresponding to different length scales is interpreted in terms of collective correlations among the particles within the assemblies

  12. The Synergistic Effect between Electrical and Chemical Factors in Plasma Gene/Molecule-Transfection

    Science.gov (United States)

    Jinno, Masafumi

    2016-09-01

    This study has been done to know what kind of factors in plasma and processes on cells promote plasma gene/molecule transfection. We have discovered a new plasma source using a microcapillary electrode which enables high transfection efficiency and high cell survivability simultaneously. However, the mechanism of the transfection by plasma was not clear. To clarify the transfection mechanisms by micro plasma, we focused on the effects of electrical (current, charge, field, etc.) and chemical (radicals, RONS, etc.) factors generated by the micro plasma and evaluated the contribution weight of three groups of the effects and processes, i.e. electrical, chemical and biochemical ones. At first, the necessity of the electrical factors was estimated by the laser produced plasma (LPP). Mouse L-929 fibroblast cell was cultured on a 96-well plate or 12-well micro slide chamber. Plasmids pCX-EGFP in Tris-EDTA buffer was dropped on the cells and they were exposed to the capillary discharge plasma (CDP) or the LPP. In the case of the CDP, the plasma was generated between the tip of the capillary electrode and the cells so that both electrical and chemical factors were supplied to the cells. In this setup, about 20% of average transfection efficiency was obtained. In the case of the LPP, the plasma was generated apart from the cells so that electrical factors were not supplied to the cells. In this setup, no transfection was observed. These results show that the electrical factors are necessary for the plasma gene transfection. Next, the necessity of the chemical factors was estimated the effect of catalase to remove H2O2 in CDP. The transfection efficiency decreased to 0.4 by scavenging H2O2 with catalase. However, only the solution of H2O2 caused no gene transfection in cells. These results shows that H2O2 is important species to cause gene/molecule transfection but still needs a synergistic effect with electrical or other chemical factors. This work was partly supported by

  13. Nanoparticles for antimicrobial purposes in Endodontics: A systematic review of in vitro studies

    Energy Technology Data Exchange (ETDEWEB)

    Samiei, Mohammad [Faculty of Dentistry, Tabriz University of Medical Sciences, Tabriz (Iran, Islamic Republic of); School of Advanced Medicine, Tabriz University of Medical Sciences, Tabriz (Iran, Islamic Republic of); Farjami, Afsaneh; Dizaj, Solmaz Maleki [Hematology & Oncology Research Center and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz (Iran, Islamic Republic of); Lotfipour, Farzaneh, E-mail: lotfipoor@tbzmed.ac.ir [School of Advanced Medicine, Tabriz University of Medical Sciences, Tabriz (Iran, Islamic Republic of); Hematology & Oncology Research Center and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz (Iran, Islamic Republic of)

    2016-01-01

    Introduction: Antimicrobial nanoparticles with enhanced physiochemical properties have attracted attention as modern antimicrobials, especially in the complicated oral cavity environment. The goal of the present article is to review the current state of nanoparticles used for antimicrobial purposes in root canal infections. Methods: A review was conducted in electronic databases using MeSH keywords to identify relevant published literature in English. The analysis and eligibility criteria were documented according to the Preferred Reporting Items for Systematic Reviews and Meta Analysis (PRISMA-guidelines). No restrictions on publication date were imposed. Data regarding root canal disinfections, general antimicrobial mechanisms of nanoparticles, type of nanoparticles as antimicrobial agent and antimicrobial effect of nanoparticles in endodontics were collected and subjected to descriptive data analysis. Results: The literature search in electronic databases according to the inclusion criteria provided 83 titles and abstracts. Among them 15 papers were related to antimicrobial effect of nanoparticles in Endodontics. Silver nanoparticles with sustainable activity were the most studied agent for its antimicrobial behavior in root canal infection. Aided polymeric nanoparticles with photo or ultrasound, glass bioactive nanoparticles as well as Calcium derivative based nanoparticles, with improved activity in comparison with the non-nano counterparts, are of importance in infection control of dental root canal. Bioactive Non-organic nanoparticles with structural capabilities present enhanced antimicrobial activity in root canal infections. Discussion: All included studies showed an enhanced or at least equal effect of nanoparticulate systems to combat dental root canal infections compared to conventional antimicrobial procedures. However, it is crucial to understand their shortcomings and their probable cellular effects and toxicity as well as environmental effects

  14. Green synthesis, morphological and optical studies of CuO nanoparticles

    Science.gov (United States)

    Reddy, K. Rayapa

    2017-12-01

    CuO nanoparticles were synthesized by environmentally benign green route with copper acetate precursor using Caloropis procera leaf extract. The CuO nanoparticles produced by the process after calcination at 700° C were characterized by X-ray diffraction (XRD) and the pattern reveals monoclinic structure. The cylindrical shape of CuO nanoparticles were observed by scanning electron microscopy. FT-IR spectra confirmed the adsorption of surfactant molecules at the surface of CuO nanoparticles and presence of Cusbnd O bonding. Thermal studies were carried out by thermogravimetric and deferential thermal analysis techniques. Raman spectra, is used to explore the local atomic arrangements and vibrations of CuO nanoparticles. In addition, UV-Visible spectra were used to estimate the band gap energy of CuO nanoparticles.

  15. Polymeric nanoparticles containing diazepam: preparation, optimization, characterization, in-vitro drug release and release kinetic study

    Science.gov (United States)

    Bohrey, Sarvesh; Chourasiya, Vibha; Pandey, Archna

    2016-03-01

    Nanoparticles formulated from biodegradable polymers like poly(lactic-co-glycolic acid) (PLGA) are being extensively investigated as drug delivery systems due to their two important properties such as biocompatibility and controlled drug release characteristics. The aim of this work to formulated diazepam loaded PLGA nanoparticles by using emulsion solvent evaporation technique. Polyvinyl alcohol (PVA) is used as stabilizing agent. Diazepam is a benzodiazepine derivative drug, and widely used as an anticonvulsant in the treatment of various types of epilepsy, insomnia and anxiety. This work investigates the effects of some preparation variables on the size and shape of nanoparticles prepared by emulsion solvent evaporation method. These nanoparticles were characterized by photon correlation spectroscopy (PCS), transmission electron microscopy (TEM). Zeta potential study was also performed to understand the surface charge of nanoparticles. The drug release from drug loaded nanoparticles was studied by dialysis bag method and the in vitro drug release data was also studied by various kinetic models. The results show that sonication time, polymer content, surfactant concentration, ratio of organic to aqueous phase volume, and the amount of drug have an important effect on the size of nanoparticles. Hopefully we produced spherical shape Diazepam loaded PLGA nanoparticles with a size range under 250 nm with zeta potential -23.3 mV. The in vitro drug release analysis shows sustained release of drug from nanoparticles and follow Korsmeyer-Peppas model.

  16. Combining polyethylenimine and Fe(III) for mediating pDNA transfection.

    Science.gov (United States)

    Jorge, Andreia F; Röder, Ruth; Kos, Petra; Dias, Rita S; Wagner, Ernst; Pais, Alberto A C C

    2015-06-01

    The potential use of Fe(III) ions in biomedical applications may predict the interest of its combination with pDNA-PEI polyplexes. The present work aims at assessing the impact of this metal on pDNA complex properties. Variations in the formation of complexes were imposed by using two types of biological buffers at different salt conditions. The incorporation of pDNA in complexes was characterised by gel electrophoresis and dynamic light scattering. Transfection efficiency and cytotoxicity were evaluated in HeLa and HUH-7 cell lines, supported by flow cytometry assays. Fe(III) enhances pDNA incorporation in the complex, irrespective of the buffer used. Transfection studies reveal that the addition of Fe(III) to complexes at low ionic strength reduces gene transfection, while those prepared under high salt content do not affect or, in a specific case, increase gene transfection up to 5 times. This increase may be a consequence of a favoured interaction of polyplexes with cell membrane and uptake. At low salt conditions, results attained with chloroquine indicate that the metal may inhibit polyplex endosomal escape. A reduction on the amount of PEI (N/P 5) formed at intermediary ionic strength, complemented by Fe(III), reduces the size of complexes while maintaining a transfection efficiency similar to that obtained to N/P 6. Fe(III) emerges as a good supporting condensing agent to modulate pDNA-PEI properties, including condensation, size and cytotoxicity, without a large penalty on gene transfection. This study highlights important aspects that govern pDNA transfection and elucidates the benefits of incorporating the versatile Fe(III) in a gene delivery system. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Transfected parvalbumin alters calcium homeostasis in teratocarcinoma PCC7 cells

    DEFF Research Database (Denmark)

    Müller, B K; Kabos, P; Belhage, B

    1996-01-01

    Indirect evidence supports a protective role of some EF-hand calcium-binding proteins against calcium-induced neurotoxicity. Little is known about how these proteins influence cytosolic calcium levels. After cloning the parvalbumin cDNA into an expression vector, teratocarcinoma cells (PCC7) were...... transfected. Parvalbumin-transfected and mock-transfected cells were loaded with the calcium indicator fura-2 and were exposed, in the same dish, to different concentrations of the calcium ionophore A23187 or to KCI. The results show that parvalbumin-transfected PCC7 cells had much better calcium buffering...

  18. PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells

    Directory of Open Access Journals (Sweden)

    Liang Gao

    2011-01-01

    Full Text Available Abstract The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly(D,L-lactide-co-glycolide (PLGA-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

  19. Comparative study of gum arabic and PVP as stabilizing agents for synthesis of gold nanoparticles

    International Nuclear Information System (INIS)

    Silva, Andressa A.; Leal, Jessica; Geraldes, Adriana N.; Lugao, Ademar B.

    2015-01-01

    Use Colloidal metallic nanoparticles such as gold nanoparticles have received a great attention, due in part to their specific properties and potential applications. Control of size and uniformity of nanoparticles is important to prevent aggregation. High-molecular-weight polymers were used as stabilizer agents. Natural polymers, such as gum Arabic, are used as stabilizer because of capping nanoparticles behavior and present advantages such as solubility, non- toxicity and its compatibility for pharmaceutical and biomedical applications. Previous studies showed that the hydrophilic group of Poly(vinyl pyrrolidone) (PVP) caused repulsion on gold nanoparticles surface because steric interactions with polymer, for this reason this kind of polymers could be used as stabilizer agent. The aim of this work is to study the synthesis and stabilization of gold nanoparticles with PVP and gum Arabic using gamma radiation. The results obtained by samples analysis using UV-Visible showed that the gamma irradiation doses influenced the nanoparticles formation by PVP but that is not the case with the GA, because for smaller quantity of Arabic gum in different doses produced and stabilized nanoparticles. The samples were observed for 20 days and showed stability. We have obtained preliminary results showed that the use of radiation is applicable to the formation of gold nanoparticles. (author)

  20. Comparative study of gum arabic and PVP as stabilizing agents for synthesis of gold nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Andressa A.; Leal, Jessica; Geraldes, Adriana N.; Lugao, Ademar B., E-mail: andressa_alvess@yahoo.com.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2015-07-01

    Use Colloidal metallic nanoparticles such as gold nanoparticles have received a great attention, due in part to their specific properties and potential applications. Control of size and uniformity of nanoparticles is important to prevent aggregation. High-molecular-weight polymers were used as stabilizer agents. Natural polymers, such as gum Arabic, are used as stabilizer because of capping nanoparticles behavior and present advantages such as solubility, non- toxicity and its compatibility for pharmaceutical and biomedical applications. Previous studies showed that the hydrophilic group of Poly(vinyl pyrrolidone) (PVP) caused repulsion on gold nanoparticles surface because steric interactions with polymer, for this reason this kind of polymers could be used as stabilizer agent. The aim of this work is to study the synthesis and stabilization of gold nanoparticles with PVP and gum Arabic using gamma radiation. The results obtained by samples analysis using UV-Visible showed that the gamma irradiation doses influenced the nanoparticles formation by PVP but that is not the case with the GA, because for smaller quantity of Arabic gum in different doses produced and stabilized nanoparticles. The samples were observed for 20 days and showed stability. We have obtained preliminary results showed that the use of radiation is applicable to the formation of gold nanoparticles. (author)

  1. A family of cationic polyamides for in vitro and in vivo gene transfection.

    Science.gov (United States)

    Zhang, Chengnan; Jin, Rong; Zhao, Peng; Lin, Chao

    2015-08-01

    The purpose of this study is to develop biodegradable cationic polyamides for non-viral gene delivery and elucidate their structural effects on gene transfection activity. To this end, a group of novel cationic polyamides were synthesized by polycondensation reaction between different di-p-nitrophenyl esters and tertiary amine-containing primary diamines. These linear polyamides have flexible alkylene group (ethylene or propylene), protonable amino group and bioreducible disulfide linkage in the polyamide main chain. The alkylene group and disulfide linkage in these polyamides have a distinct effect on their gene delivery properties including buffering capacity, gene binding ability and intracellular gene release profile. Those cationic polyamides containing disulfide linkage and 1,4-bis(3-aminopropyl)piperazine (BAP) residue exhibited high buffering capacity (endosomal escape ability), high gene binding ability, and intracellular gene release ability, thus inducing fast gene nucleus translocation and robust gene transfection in vitro against different cell lines and rat bone marrow mesenchymal stem cells. Moreover, the transfection efficiencies in vitro were comparable or higher than those of 25 kDa branched polyethylenimine and Lipofectamine 2000 transfection agent as positive controls. These cationic polyamides and their polyplexes were of low cytotoxicity when an optimal transfection efficacy was achieved. In vivo transfection tests showed that bioreducible BAP-based polyamides were applicable for intravenous gene delivery in a mouse model, leading to higher level of transgene expression in the liver as compared to 22 kDa linear polyethylenimine as a positive control. These cationic polyamides provide a useful platform to elucidate the relationship between chemical functionalities and gene transfection activity. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  2. Chitosan polyplex nanoparticle vector for miR-145 expression in MCF-7: Optimization by design of experiment.

    Science.gov (United States)

    Tekie, Farnaz Sadat Mirzazadeh; Atyabi, Fatemeh; Soleimani, Masoud; Arefian, Ehsan; Atashi, Amir; Kiani, Melika; Khoshayand, Mohammad Reza; Amini, Mohsen; Dinarvand, Rassoul

    2015-11-01

    miR-145, a tumor suppressor micro RNA (miRNA), is down regulated in cancer and can be introduced as a therapeutic agent in various cancers including breast cancer. In this study, miR-145 plasmid was transfected to MCF-7 cells using chitosan polyplex nanoparticles. The vector was prepared according to an optimized fabricating method determined by response surface analysis and D-optimal design. Effects of chitosan molecular weight (Mw) and polymer amine to DNA phosphate ratio (N/P) as the variables were investigated on size, zeta potential, stability, and transfection efficiency of the polyplex nanoparticles. The results indicated that there is an interaction between effects of Mw and N/P ratio on the size of nanoparticles. Gel retardation assay demonstrated that the stability of the complexes in serum and preparation medium during storage time depends on the formulation variables. Statistical analysis affirmed that in spite of particle size, the variables of N/P ratio, time of incubation, and zeta potential affect the gene transfection. In conclusion, by selecting the perfect formulation prepared through an optimized method, it is possible to achieve a high transfection efficacy for miR-145 as an anticancer biological macromolecule. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. A rapid pathway toward a superb gene delivery system: programming structural and functional diversity into a supramolecular nanoparticle library.

    Science.gov (United States)

    Wang, Hao; Liu, Kan; Chen, Kuan-Ju; Lu, Yujie; Wang, Shutao; Lin, Wei-Yu; Guo, Feng; Kamei, Ken-ichiro; Chen, Yi-Chun; Ohashi, Minori; Wang, Mingwei; Garcia, Mitch André; Zhao, Xing-Zhong; Shen, Clifton K-F; Tseng, Hsian-Rong

    2010-10-26

    Nanoparticles are regarded as promising transfection reagents for effective and safe delivery of nucleic acids into a specific type of cells or tissues providing an alternative manipulation/therapy strategy to viral gene delivery. However, the current process of searching novel delivery materials is limited due to conventional low-throughput and time-consuming multistep synthetic approaches. Additionally, conventional approaches are frequently accompanied with unpredictability and continual optimization refinements, impeding flexible generation of material diversity creating a major obstacle to achieving high transfection performance. Here we have demonstrated a rapid developmental pathway toward highly efficient gene delivery systems by leveraging the powers of a supramolecular synthetic approach and a custom-designed digital microreactor. Using the digital microreactor, broad structural/functional diversity can be programmed into a library of DNA-encapsulated supramolecular nanoparticles (DNA⊂SNPs) by systematically altering the mixing ratios of molecular building blocks and a DNA plasmid. In vitro transfection studies with DNA⊂SNPs library identified the DNA⊂SNPs with the highest gene transfection efficiency, which can be attributed to cooperative effects of structures and surface chemistry of DNA⊂SNPs. We envision such a rapid developmental pathway can be adopted for generating nanoparticle-based vectors for delivery of a variety of loads.

  4. [Comparison of efficiency and cytotoxicity of different transfection reagents in transfecting RIP140-siRNA into Kupffer cells].

    Science.gov (United States)

    Li, Ji; Liu, Zuojin

    2015-12-01

    To compare the efficiency and cytotoxicity of different transfection reagents used in transfection of RIP140-siRNA into Kupffer cells to optimize the transfection conditions. Kupffer cells were transfected with RIP140-siRNA labeled with GFP as the reporter gene using lipofectamine 2000, Roche reagent (X-treme GENE siRNA Transfection Reagent) and puro screening lentivirus (1.0×10(8) TU/mL) as the transfection reagents. The transfection effect was observed under a fluorescent inverted microscope, and laser scanning confocal microscopy was used to analyze RIP140 expression in trasnfected Kupffer cells. Flow cytometry was performed to detect cell apoptosis, and CCK-8 test was used to evaluate the cell proliferation inhibition. RT-RCR and Western blotting were performed to detect the expressions of RIP140 mRNA and protein in the trasnfected cells. Puro screening lentivirus yielded the highest cell transfection efficiency, which exceeded 90%, followed by Roche reagent and then by lipofectamine 2000. Flow cytometry and CCK-8 test showed that the cytotoxicity was the mildest with Roche reagent, moderate with lentivirus, and severe with lipofectamine 2000. The cells trasnfected with lentivirus showed a significantly lower RIP140 expression than cells trasnfected with lipofectamine 2000 and Roche reagent (Ptransfection, as compared with the other two trasnfection reagents, can achieve good transfection efficiency with a relativelty low cytotoxicity, and allows for better controllability and stability of the trasnfectiion conditions.

  5. In vivo toxicologic study of larger silica nanoparticles in mice

    Directory of Open Access Journals (Sweden)

    Chan WT

    2017-04-01

    Full Text Available Wai-Tao Chan,1–3 Cheng-Che Liu,4 Jen-Shiu Chiang Chiau,5 Shang-Ting Tsai,6 Chih-Kai Liang,6 Mei-Lien Cheng,5 Hung-Chang Lee,7,8 Chun-Yun Yeung,1,3,9 Shao-Yi Hou2,6 1Department of Pediatric Gastroenterology, Hepatology and Nutrition, MacKay Children’s Hospital, 2Graduate Institute of Engineering Technology, National Taipei University of Technology, 3Mackay Medicine, Nursing, and Management College, 4Institute of Preventive Medicine, National Defense Medical Center, Taipei, 5Department of Medical Research, MacKay Memorial Hospital, Hsinchu, 6Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei, 7Department of Pediatrics, MacKay Memorial Hospital, Hsinchu, 8Department of Pediatrics, Taipei Medical University, Taipei, 9Department of Medicine, Mackay Medical College, New Taipei, Taiwan, Republic of China Abstract: Silica nanoparticles (SiNPs are being studied and used for medical purposes. As nanotechnology grows rapidly, its biosafety and toxicity have frequently raised concerns. However, diverse results have been reported about the safety of SiNPs; several studies reported that smaller particles might exhibit toxic effects to some cell lines, and larger particles of 100 nm were reported to be genotoxic to the cocultured cells. Here, we investigated the in vivo toxicity of SiNPs of 150 nm in various dosages via intravenous administration in mice. The mice were observed for 14 days before blood examination and histopathological assay. All the mice survived and behaved normally after the administration of nanoparticles. No significant weight change was noted. Blood examinations showed no definite systemic dysfunction of organ systems. Histopathological studies of vital organs confirmed no SiNP-related adverse effects. We concluded that 150 nm SiNPs were biocompatible and safe for in vivo use in mice. Keywords: in vivo, mice, silica nanoparticle, nanotoxicity

  6. Gene therapy for C-26 colon cancer using heparin-polyethyleneimine nanoparticle-mediated survivin T34A

    Directory of Open Access Journals (Sweden)

    Zhang L

    2011-10-01

    Full Text Available Ling Zhang1,*, Xiang Gao1,2,*, Ke Men1, BiLan Wang1, Shuang Zhang1, Jinfeng Qiu1, Meijuan Huang1, MaLing Gou1, Ning Huang2, ZhiYong Qian1, Xia Zhao1, YuQuan Wei11State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, 2Department of Pathophysiology, College of Preclinical and Forensic Medical Sciences, Sichuan University, Chengdu, People’s Republic of China*These authors contributed equally to this workBackground: Gene therapy provides a novel method for the prevention and treatment of cancer, but the clinical application of gene therapy is restricted, mainly because of the absence of an efficient and safe gene delivery system. Recently, we developed a novel nonviral gene carrier, ie, heparin-polyethyleneimine (HPEI nanoparticles for this purpose.Methods and results: HPEI nanoparticles were used to deliver plasmid-expressing mouse survivin-T34A (ms-T34A to treat C-26 carcinoma in vitro and in vivo. According to the in vitro studies, HPEI nanoparticles could efficiently transfect the pGFP report gene into C-26 cells, with a transfection efficiency of 30.5% ± 2%. Moreover, HPEI nanoparticle-mediated ms-T34A could efficiently inhibit the proliferation of C-26 cells by induction of apoptosis in vitro. Based on the in vivo studies, HPEI nanoparticles could transfect the Lac-Z report gene into C-26 cells in vivo. Intratumoral injection of HPEI nanoparticle-mediated ms-T34A significantly inhibited growth of subcutaneous C-26 carcinoma in vivo by induction of apoptosis and inhibition of angiogenesis.Conclusion: This research suggests that HPEI nanoparticle-mediated ms-T34A may have a promising role in C-26 colon carcinoma therapy.Keywords: gene therapy, mouse survivin-T34A, colon cancer, polyethyleneimine, nanoparticles, cancer therapy

  7. Cationic, amphiphilic copolymer micelles as nucleic acid carriers for enhanced transfection in rat spinal cord.

    Science.gov (United States)

    Gwak, So-Jung; Nice, Justin; Zhang, Jeremy; Green, Benjamin; Macks, Christian; Bae, Sooneon; Webb, Ken; Lee, Jeoung Soo

    2016-04-15

    Spinal cord injury commonly leads to permanent motor and sensory deficits due to the limited regenerative capacity of the adult central nervous system (CNS). Nucleic acid-based therapy is a promising strategy to deliver bioactive molecules capable of promoting axonal regeneration. Branched polyethylenimine (bPEI: 25kDa) is one of the most widely studied nonviral vectors, but its clinical application has been limited due to its cytotoxicity and low transfection efficiency in the presence of serum proteins. In this study, we synthesized cationic amphiphilic copolymers, poly (lactide-co-glycolide)-graft-polyethylenimine (PgP), by grafting low molecular weight PLGA (4kDa) to bPEI (25kDa) at approximately a 3:1 ratio as an efficient nonviral vector. We show that PgP micelle is capable of efficiently transfecting plasmid DNA (pDNA) and siRNA in the presence of 10% serum in neuroglioma (C6) cells, neuroblastoma (B35) cells, and primary E8 chick forebrain neurons (CFN) with pDNA transfection efficiencies of 58.8%, 75.1%, and 8.1%, respectively. We also show that PgP provides high-level transgene expression in the rat spinal cord in vivo that is substantially greater than that attained with bPEI. The combination of improved transfection and reduced cytotoxicity in vitro in the presence of serum and in vivo transfection of neural cells relative to conventional bPEI suggests that PgP may be a promising nonviral vector for therapeutic nucleic acid delivery for neural regeneration. Gene therapy is a promising strategy to overcome barriers to axonal regeneration in the injured central nervous system. Branched polyethylenimine (bPEI: 25kDa) is one of the most widely studied nonviral vectors, but its clinical application has been limited due to cytotoxicity and low transfection efficiency in the presence of serum proteins. Here, we report cationic amphiphilic copolymers, poly (lactide-co-glycolide)-graft-polyethylenimine (PgP) that are capable of efficiently transfecting reporter

  8. Transfection of brain capillary endothelial cells in primary culture with defined blood-brain barrier properties.

    Science.gov (United States)

    Burkhart, Annette; Thomsen, Louiza Bohn; Thomsen, Maj Schneider; Lichota, Jacek; Fazakas, Csilla; Krizbai, István; Moos, Torben

    2015-08-07

    Primary brain capillary endothelial cells (BCECs) are a promising tool to study the blood-brain barrier (BBB) in vitro, as they maintain many important characteristics of the BBB in vivo, especially when co-cultured with pericytes and/or astrocytes. A novel strategy for drug delivery to the brain is to transform BCECs into protein factories by genetic modifications leading to secretion of otherwise BBB impermeable proteins into the central nervous system. However, a huge challenge underlying this strategy is to enable transfection of non-mitotic BCECs, taking a non-viral approach. We therefore aimed to study transfection in primary, non-mitotic BCECs cultured with defined BBB properties without disrupting the cells' integrity. Primary cultures of BCECs, pericytes and astrocytes were generated from rat brains and used in three different in vitro BBB experimental arrangements, which were characterised based on a their expression of tight junction proteins and other BBB specific proteins, high trans-endothelial electrical resistance (TEER), and low passive permeability to radiolabeled mannitol. Recombinant gene expression and protein synthesis were examined in primary BCECs. The BCECs were transfected using a commercially available transfection agent Turbofect™ to express the red fluorescent protein HcRed1-C1. The BCECs were transfected at different time points to monitor transfection in relation to mitotic or non-mitotic cells, as indicated by fluorescence-activated cell sorting analysis after 5-and 6-carboxylfluorescein diacetate succinidyl ester incorporation. The cell cultures exhibited important BBB characteristics judged from their expression of BBB specific proteins, high TEER values, and low passive permeability. Among the three in vitro BBB models, co-culturing with BCECs and astrocytes was well suited for the transfection studies. Transfection was independent of cell division and with equal efficacy between the mitotic and non-mitotic BCECs. Importantly

  9. In vitro transfection of bone marrow-derived dendritic cells with TATp-liposomes.

    Science.gov (United States)

    Pappalardo, Juan Sebastián; Langellotti, Cecilia A; Di Giacomo, Sebastián; Olivera, Valeria; Quattrocchi, Valeria; Zamorano, Patricia I; Hartner, William C; Levchenko, Tatyana S; Torchilin, Vladimir P

    2014-01-01

    Dendritic cells (DC) are antigen-presenting cells uniquely capable of priming naïve T cells and cross-presenting antigens, and they determine the type of immune response elicited against an antigen. TAT peptide (TATp), is an amphipathic, arginine-rich, cationic peptide that promotes penetration and translocation of various molecules and nanoparticles into cells. TATp-liposomes (TATp-L) used for DC transfection were prepared using TATp derivatized with a lipid-terminated polymer capable of anchoring in the liposomal membrane. Here, we show that the addition of TATp to DNA-loaded liposomes increased the uptake of DNA in DC. DNA-loaded TATp-L increased the in vitro transfection efficiency in DC cultures as evidenced by a higher expression of the enhanced green fluorescent protein and bovine herpes virus type 1 glycoprotein D (gD). The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L, as indicated by the secretion of interleukin 6.

  10. Drug Delivery and Cell Transfection Using Shock Waves Produced by Nanothermites

    Science.gov (United States)

    Gangopadhyay, Shubhra

    2009-06-01

    Shock waves have non-destructive life science applications in cell transfection and drug delivery. Based on molecular dynamics simulations, the shockwave causes transient compression of the cell membrane, which causes the hydrophobic interior of the lipid bilayer to become thinner. This allows diffusion of water molecules across the membrane. Recently, the nanothermite composition consisting of CuO nanorods and Al nanoparticles was shown to propagate at velocities in the same range as metallic azides and fulminates; however, the CuO/Al mixture produces lower pressure levels. An in vitro testing system was developed to deliver shock waves produced by nanothermites into cell suspensions and/or tissues. The plasmid encoded for production of green-fluorescent protein was delivered into cells including, among other types, chicken cardiomyocytes, cell lines (T47-D and Ins-1), and Arabidopsis plant cells. It was found that the nanothermite pressure impulses induced transfection resulting in production of green fluorescent protein in 99% of the cardiomyocytes. Additionally, transfected cell survival was evaluated, and the pressure impulses did not produce any elevated levels of cell death compared with control cell suspensions.

  11. The challenges of testing metal and metal oxide nanoparticles in algal bioassays: titanium dioxide and gold nanoparticles as case studies

    DEFF Research Database (Denmark)

    Hartmann, Nanna Isabella Bloch; Engelbrekt, Christian; Zhang, Jingdong

    2013-01-01

    Aquatic toxicology of engineered nanoparticles is challenged by methodological difficulties stemming partly from highly dynamic and poorly understood behavior of nanoparticles in biological test systems. In this paper scientific and technical challenges of testing not readily soluble nanoparticle...

  12. Multifunctional non-viral gene vectors with enhanced stability, improved cellular and nuclear uptake capability, and increased transfection efficiency

    Science.gov (United States)

    Yang, Zhe; Jiang, Zhaozhong; Cao, Zhong; Zhang, Chao; Gao, Di; Luo, Xingen; Zhang, Xiaofang; Luo, Huiyan; Jiang, Qing; Liu, Jie

    2014-08-01

    We have developed a new multifunctional, non-viral gene delivery platform consisting of cationic poly(amine-co-ester) (PPMS) for DNA condensation, PEG shell for nanoparticle stabilization, poly(γ-glutamic acid) (γ-PGA) and mTAT (a cell-penetrating peptide) for accelerated cellular uptake, and a nuclear localization signal peptide (NLS) for enhanced intracellular transport of DNA to the nucleus. In vitro study showed that coating of the binary PPMS/DNA polyplex with γ-PGA promotes cellular uptake of the polyplex particles, particularly by γ-glutamyl transpeptidase (GGT)-positive cells through the GGT-mediated endocytosis pathway. Conjugating PEG to the γ-PGA led to the formation of a ternary PPMS/DNA/PGA-g-PEG polyplex with decreased positive charges on the surface of the polyplex particles and substantially higher stability in serum-containing aqueous medium. The cellular uptake rate was further improved by incorporating mTAT into the ternary polyplex system. Addition of the NLS peptide was designed to facilitate intracellular delivery of the plasmid to the nucleus--a rate-limiting step in the gene transfection process. As a result, compared with the binary PPMS/LucDNA polyplex, the new mTAT-quaternary PPMS/LucDNA/NLS/PGA-g-PEG-mTAT system exhibited reduced cytotoxicity, remarkably faster cellular uptake rate, and enhanced transport of DNA to the nucleus. All these advantageous functionalities contribute to the remarkable gene transfection efficiency of the mTAT-quaternary polyplex both in vitro and in vivo, which exceeds that of the binary polyplex and commercial Lipofectamine™ 2000/DNA lipoplex. The multifunctional mTAT-quaternary polyplex system with improved efficiency and reduced cytotoxicity represents a new type of promising non-viral vectors for the delivery of therapeutic genes to treat tumors.We have developed a new multifunctional, non-viral gene delivery platform consisting of cationic poly(amine-co-ester) (PPMS) for DNA condensation, PEG shell

  13. A study of phase transfer processes of Ag nanoparticles

    Science.gov (United States)

    Li, De-Gang; Chen, Shen-Hao; Zhao, Shi-Yong; Hou, Xian-Ming; Ma, Hou-Yi; Yang, Xue-Geng

    2002-11-01

    With the protection of sodium oleate, Ag nanoparticles are produced through the reduction of AgNO 3 with NaBH 4 in an aqueous solution. The possible mechanism of phase transfer of the Ag nanoparticles was discussed. At a suitable concentration of sodium oleate, after adding NaH 2PO 4, the oleic acid molecule can change its position on the surface of Ag nanoparticles under the effects of water and toluene and become amphipathic. So most of the nanoparticles form a film between water/toluene. For the case of a higher concentration of sodium oleate, excess sodium oleate will form a closed monolayer film on the surface of the Ag nanoparticles. After adding NaH 2PO 4, the oleic acid molecule cannot move on the Ag nanoparticles surface, thus the colloid particles are hydrophobic but not amphipathic. So most of the particles transfer to the organic phase. UV-Vis spectra, TEM and conventional metallographic microscopy are used to characterize the Ag nanoparticles and nanoparticles films.

  14. Experimental and theoretical studies of nanoparticles of antiferromagnetic materials

    DEFF Research Database (Denmark)

    Mørup, Steen; Madsen, Daniel Esmarch; Frandsen, Cathrine

    2007-01-01

    The magnetic properties of nanoparticles of antiferromagnetic materials are reviewed. The magnetic structure is often similar to the bulk structure, but there are several examples of size-dependent magnetic structures. Owing to the small magnetic moments of antiferromagnetic nanoparticles, the co...

  15. Mössbauer studies of superparamagnetic ferrite nanoparticles for functional application

    International Nuclear Information System (INIS)

    Mažeika, K.; Jagminas, A.; Kurtinaitienė, M.

    2013-01-01

    Nanoparticles of CoFe 2 O 4 and MnFe 2 O 4 prepared for functional applications in nanomedicine were studied using Mössbauer spectrometry. Superparamagnetic properties of nanoparticles of different size and composition were compared applying collective excitations and multilevel models for the description of the Mössbauer spectra.

  16. Complexity of gold nanoparticle formation disclosed by dynamics study

    DEFF Research Database (Denmark)

    Engelbrekt, Christian; Jensen, Palle Skovhus; Sørensen, Karsten

    2013-01-01

    Although chemically synthesized gold nanoparticles (AuNPs) from gold salt (HAuCl4) are among the most studied nanomaterials, understanding the formation mechanisms is a challenge mainly due to limited dynamics information. A range of in situ methods with down to millisecond (ms) time resolution...... have been employed in the present report to monitor time-dependent physical and chemical properties in aqueous solution during the chemical synthesis. Chemical synthesis of AuNPs is a reduction process accompanied by release of ions and protons, and formation of solid particles. Dynamic information...... from redox potential, pH, conductivity, and turbidity of the solution enables distinct observation of reduction and nucleation/growth of AuNPs phases. The dynamics of the electrochemical potential shows that reduction of gold salt (HAuCl 4 and its hydrolyzed forms) occurs via intermediate [AuCl 2...

  17. Density functional theory studies of transition metal nanoparticles in catalysis

    DEFF Research Database (Denmark)

    Greeley, Jeffrey Philip; Rankin, Rees; Zeng, Zhenhua

    2013-01-01

    Periodic Density Functional Theory calculations are capable of providing powerful insights into the structural, energetics, and electronic phenomena that underlie heterogeneous catalysis on transition metal nanoparticles. Such calculations are now routinely applied to single crystal metal surfaces...... and to subnanometer metal clusters. Descriptions of catalysis on truly nanosized structures, however, are generally not as well developed. In this talk, I will illustrate different approaches to analyzing nanocatalytic phenomena with DFT calculations. I will describe case studies from heterogeneous catalysis...... and electrocatalysis, in which single crystal models are combined with Wulff construction-based ideas to produce descriptions of average nanocatalyst behavior. Then, I will proceed to describe explicitly DFT-based descriptions of catalysis on truly nanosized particles (

  18. Correlation between cationic lipid-based transfection and cell division

    Energy Technology Data Exchange (ETDEWEB)

    Kirchenbuechler, Inka; Kirchenbuechler, David; Elbaum, Michael, E-mail: michael@elbaum.ac.il

    2016-07-01

    We evaluate the temporal relation between protein expression by cationic lipid-mediated transfection and cell division using time lapse fluorescence microscopy. Detailed image analysis provides new insights on the single cell level while simultaneously achieving appropriate statistics. Earlier evidence by less direct methods such as flow cytometry indicates a primary route for transfection involving nuclear envelope breakdown, but also suggests the existence of a pathway independent of mitosis. We confirm and quantify both mechanisms. We found the timing for successful transfection to be unexpectedly flexible, contrary to assertions of a narrow time window. Specifically, cells dividing more than 24 h after exposure to the transfection medium express the probed protein at a comparable level to cells in a mitotic state during or shortly after transfection. This finding can have a profound impact on the guidance and development of non-viral gene delivery materials. - Highlights: • Cationic lipid-based transfection supports protein expression without cell division. • Protein expression is unrelated to cell cycle status at the time of transfection. • Time-lapse imaging provides direct evaluation without statistical averaging. • Lipoplex dissociation is a likely target for improvement of transfection efficiency.

  19. A nanoparticle dispersion method for in vitro and in vivo nanotoxicity study.

    Science.gov (United States)

    Kim, Seong C; Chen, Da-Ren; Qi, Chaolong; Gelein, Robert M; Finkelstein, Jacob N; Elder, Alison; Bentley, Karen; Oberdörster, Günter; Pui, David Y H

    2010-03-01

    The dispersion in air of nanoparticles of different sizes, materials and morphologies with controlled agglomeration involving aerosol delivery for in vivo and in vitro studies is one of the most difficult challenges in the field of nanoparticle toxicology. We describe here a nanoparticle dispersion system using an electrospray method to deliver airborne nanoparticles (approximately 10-100 nm) with spatial uniformity and controllable particle concentration for in vitro and in vivo studies. With the dispersion method, single nanoparticles (polystyrene latex particles, TiO(2), Au, Mn, quantum dots, and carbon nanotubes) can be delivered to cells and animals via the air. The degree of agglomeration can be controlled by changing the suspension feeding rate to simulate realistic conditions for exposure studies.

  20. [EFFECT OF TRITON X-100 ON LIPOSOME MEDIATED BONE MORPHOGENETIC PROTEIN 2 BY TRANSFECTION OF RAT BONE MARROW MESENCHYMAL STEM CELLS].

    Science.gov (United States)

    Xia, Delin; Huang, Mingke; Fu, Guangxing; Ma, Zheng; Wu, Shuangjiang; Zhou, Hangyu

    2015-01-01

    To study the effect of Triton X-100 promoting liposome-mediated bone morphogenetic protein 2 (BMP-2) gene transfection of rat bone marrow mesenchymal stem cells (BMSCs). BMSCs were separated and cultured from the femur and tibia of healthy Wistar rats (8-week-old, male). The 3rd passage BMSCs identified by detecting the surface antigen were used to transfect. The optimum concentration of Triton X-100 for liposome mediated gene transfection was determined with ELISA meter by the way of MTT. In optimum concentration of Triton X-100, liposome mediated BMP-2 gene was transfected to BMSCs. The experiment was divided into 3 groups according to types of trasfection agents: BMSCs were transfected with Triton X-100+liposome+BMP-2 (experimental group), with liposome+ BMP-2 (conventional transfection group), and untransfected BMSCs served as blank control group. After 48 hours of transfecting, the green fluorescent protein (GFP) in cells was detected through inverted fluorescence microscope. After 72 hours of transfection, real-time fluorescence quantitative PCR was applied to measure the mRNA expression of BMP-2. 0.01% Triton X-100 was determined to be the optimum concentration for not only making the BMSCs maintain vitality, but also achieving a certain effect on BMSCs. After trasfecting for 48 hours, GFP was observed through inverted fluorescence microscope in the experimental group and conventional transfection group, but was not observed in the blank control group. After trasfecting for 72 hours, the relative BMP-2 mRNA expression level was 5.94 ± 0.12 in the experimental group, and was 4.99 ± 0.08 in the conventional transfection group, showing significant difference (t = 360.28, P = 0.02). The transfection efficiency was increased by 19% in the experimental group. 0.010% Triton X-100 can promote the liposome mediated BMP-2 gene transfection of rat BMSUs, and can improve the transfection efficiency.

  1. Nanoparticles for antimicrobial purposes in Endodontics: A systematic review of in vitro studies.

    Science.gov (United States)

    Samiei, Mohammad; Farjami, Afsaneh; Dizaj, Solmaz Maleki; Lotfipour, Farzaneh

    2016-01-01

    Antimicrobial nanoparticles with enhanced physiochemical properties have attracted attention as modern antimicrobials, especially in the complicated oral cavity environment. The goal of the present article is to review the current state of nanoparticles used for antimicrobial purposes in root canal infections. A review was conducted in electronic databases using MeSH keywords to identify relevant published literature in English. The analysis and eligibility criteria were documented according to the Preferred Reporting Items for Systematic Reviews and Meta Analysis (PRISMA-guidelines). No restrictions on publication date were imposed. Data regarding root canal disinfections, general antimicrobial mechanisms of nanoparticles, type of nanoparticles as antimicrobial agent and antimicrobial effect of nanoparticles in endodontics were collected and subjected to descriptive data analysis. The literature search in electronic databases according to the inclusion criteria provided 83 titles and abstracts. Among them 15 papers were related to antimicrobial effect of nanoparticles in Endodontics. Silver nanoparticles with sustainable activity were the most studied agent for its antimicrobial behavior in root canal infection. Aided polymeric nanoparticles with photo or ultrasound, glass bioactive nanoparticles as well as Calcium derivative based nanoparticles, with improved activity in comparison with the non-nano counterparts, are of importance in infection control of dental root canal. Bioactive Non-organic nanoparticles with structural capabilities present enhanced antimicrobial activity in root canal infections. All included studies showed an enhanced or at least equal effect of nanoparticulate systems to combat dental root canal infections compared to conventional antimicrobial procedures. However, it is crucial to understand their shortcomings and their probable cellular effects and toxicity as well as environmental effects. Copyright © 2015 Elsevier B.V. All rights

  2. The development of mechanically formed stable nanobubbles intended for sonoporation-mediated gene transfection.

    Science.gov (United States)

    Abdalkader, Rodi; Kawakami, Shigeru; Unga, Johan; Higuchi, Yuriko; Suzuki, Ryo; Maruyama, Kazuo; Yamashita, Fumiyoshi; Hashida, Mitsuru

    2017-11-01

    In this study, stable nano-sized bubbles (nanobubbles [NBs]) were produced using the mechanical agitation method in the presence of perfluorocarbon gases. NBs made with perfluoropropane had a smaller size (around 400 nm) compared to that of those made with perfluorobutane or nitrogen gas. The lipid concentration in NBs affected both their initial size and post-formulation stability. NBs formed with a final lipid concentration of 0.5 mg/ml tended to be more stable, having a uniform size distribution for 24 h at room temperature and 50 h at 4 °C. In vitro gene expression revealed that NBs/pDNA in combination with ultrasound (US) irradiation had significantly higher transfection efficacy in colon C26 cells. Moreover, for in vivo gene transfection in mice left limb muscles, there was notable local transfection activity by NBs/pDNA when combined with US irradiation. In addition, the aged NBs kept at room temperature or 4 °C were still functional at enhancing gene transfection in mice. We succeeded in preparing stable NBs for efficient in vivo gene transfection, using the mechanical agitation method.

  3. Proton NMR studies of functionalized nanoparticles in aqueous environments

    Science.gov (United States)

    Tataurova, Yulia Nikolaevna

    Nanoscience is an emerging field that can provide potential routes towards addressing critical issues such as clean and sustainable energy, environmental remediation and human health. Specifically, porous nanomaterials, such as zeolites and mesoporous silica, are found in a wide range of applications including catalysis, drug delivery, imaging, environmental protection, and sensing. The characterization of the physical and chemical properties of nanocrystalline materials is essential to the realization of these innovative applications. The great advantage of porous nanocrystals is their increased external surface area that can control their biological, chemical and catalytic activities. Specific functional groups synthesized on the surface of nanoparticles are able to absorb heavy metals from the solution or target disease cells, such as cancer cells. In these studies, three main issues related to functionalized nanomaterials will be addressed through the application of nuclear magnetic resonance (NMR) techniques including: 1) surface composition and structure of functionalized nanocrystalline particles; 2) chemical properties of the guest molecules on the surface of nanomaterials, and 3) adsorption and reactivity of surface bound functional groups. Nuclear magnetic resonance (NMR) is one of the major spectroscopic techniques available for the characterization of molecular structure and conformational dynamics with atomic level detail. This thesis deals with the application of 1H solution state NMR to porous nanomaterial in an aqueous environment. Understanding the aqueous phase behavior of functionalized nanomaterials is a key factor in the design and development of safe nanomaterials because their interactions with living systems are always mediated through the aqueous phase. This is often due to a lack of fundamental knowledge in interfacial chemical and physical phenomena that occur on the surface of nanoparticles. The use of solution NMR spectroscopy results

  4. Functionalized polystyrene nanoparticles as a platform for studying bio–nano interactions

    Directory of Open Access Journals (Sweden)

    Cornelia Loos

    2014-12-01

    Full Text Available Nanoparticles of various shapes, sizes, and materials carrying different surface modifications have numerous technological and biomedical applications. Yet, the mechanisms by which nanoparticles interact with biological structures as well as their biological impact and hazards remain poorly investigated. Due to their large surface to volume ratio, nanoparticles usually exhibit properties that differ from those of bulk materials. Particularly, the surface chemistry of the nanoparticles is crucial for their durability and solubility in biological media as well as for their biocompatibility and biodistribution. Polystyrene does not degrade in the cellular environment and exhibits no short-term cytotoxicity. Because polystyrene nanoparticles can be easily synthesized in a wide range of sizes with distinct surface functionalizations, they are perfectly suited as model particles to study the effects of the particle surface characteristics on various biological parameters. Therefore, we have exploited polystyrene nanoparticles as a convenient platform to study bio–nano interactions. This review summarizes studies on positively and negatively charged polystyrene nanoparticles and compares them with clinically used superparamagnetic iron oxide nanoparticles.

  5. Correlation between cationic lipid-based transfection and cell division.

    Science.gov (United States)

    Kirchenbuechler, Inka; Kirchenbuechler, David; Elbaum, Michael

    2016-07-01

    We evaluate the temporal relation between protein expression by cationic lipid-mediated transfection and cell division using time lapse fluorescence microscopy. Detailed image analysis provides new insights on the single cell level while simultaneously achieving appropriate statistics. Earlier evidence by less direct methods such as flow cytometry indicates a primary route for transfection involving nuclear envelope breakdown, but also suggests the existence of a pathway independent of mitosis. We confirm and quantify both mechanisms. We found the timing for successful transfection to be unexpectedly flexible, contrary to assertions of a narrow time window. Specifically, cells dividing more than 24h after exposure to the transfection medium express the probed protein at a comparable level to cells in a mitotic state during or shortly after transfection. This finding can have a profound impact on the guidance and development of non-viral gene delivery materials. Copyright © 2016. Published by Elsevier Inc.

  6. Purification of DNA for the transfection of a Spodoptera frugiperda cell line.

    Science.gov (United States)

    Slack, Jeffrey M; Lawrence, Susan D

    2002-01-01

    Spodoptera frugiperda (Sf-9) cells have been widely used in baculovirus expression systems, transient gene expression studies and transgenic cell lines. These applications commonly require the transfection of bacterial plasmid DNA. One of the most reliable methods of preparing transfection-quality plasmid DNA is cesium chloride (CsCl) density gradient centrifugation. However, the traditional CsCl DNA purification is a long and laborious process. We have made a series of modifications to the traditional method that makes it faster, safer and easier. In the current study we demonstrate that DNA prepared by our modified CsCl method was also better for the transfection of Sf-9 cells than DNA prepared by the traditional CsCl method.

  7. Anionic solid lipid nanoparticles supported on protamine/DNA complexes

    International Nuclear Information System (INIS)

    Ye Jiesheng; Liu Chunxi; Chen Zhijin; Zhang Na; Wang Aihua

    2008-01-01

    The objective of this study was to design novel anionic ternary nanoparticles for gene delivery. These ternary nanoparticles were equipped with protamine/DNA binary complexes (150-200 nm) as the support, and the anionic formation was achieved by absorption of anionic solid lipid nanoparticles (≤20 nm) onto the surface of the binary complexes. The small solid lipid nanoparticles (SLNs) were prepared by a modified film dispersion-ultrasonication method, and adsorption of the anionic SLNs onto the binary complexes was typically carried out in water via electrostatic interaction. The formulated ternary nanoparticles were found to be relatively uniform in size (257.7 ± 10.6 nm) with a 'bumpy' surface, and the surface charge inversion from 19.28 ± 1.14 mV to -17.16 ± 1.92 mV could be considered as evidence of the formation of the ternary nanoparticles. The fluorescence intensity measurements from three batches of the ternary nanoparticles gave a mean adsorption efficiency of 96.75 ± 1.13%. Circular dichroism spectra analysis showed that the protamine/DNA complexes had been coated by small SLNs, and that the anionic ternary nanoparticles formed did not disturb the construction of the binary complexes. SYBR Green I analysis suggested that the ternary nanoparticles could protect the DNA from nuclease degradation, and cell viability assay results showed that they exhibit lower cytotoxicity to A549 cells compared with the binary complexes and lipofectamine. The transfection efficiency of the ternary nanoparticles was better than that of naked DNA and the binary complexes, and almost equal to that of lipofectamine/DNA complexes, as revealed by inversion fluorescence microscope observation. These results indicated that the anionic ternary nanoparticles could facilitate gene transfer in cultured cells, and might alleviate the drawbacks of the conventional cationic vector/DNA complexes for gene delivery in vivo

  8. Electron transfer at the contact between Al electrode and gold nanoparticles of polymer: Nanoparticle resistive switching devices studied by alternating current impedance spectroscopy

    International Nuclear Information System (INIS)

    Ouyang, Jianyong

    2013-01-01

    Electron transfer at the contact between an Al electrode and Au nanoparticles of polymer:nanoparticle devices is studied by ac impedance spectroscopy. The devices have a polystyrene layer embedded with Au nanoparticles capped with conjugated 2-naphthalenethiol sandwiched between Al and MoO 3 /Al electrodes, and they exhibit electrode-sensitive resistive switches. The devices in the pristine or high resistance state have high capacitance. The capacitance decreases after the devices switch to a low resistance state by a voltage scan. The change in the capacitance is attributed to the voltage-induced change on the electronic structure of the contact between the Al electrode and Au nanoparticles

  9. Combined TEM and NC-AFM study of Al2O3-supported Pt nanoparticles

    DEFF Research Database (Denmark)

    Jensen, Thomas Nørregaard; Simonsen, Søren Bredmose; Chorkendorff, Ib

    Sintering, the growth of large particles at the expense of smaller ones, is one of the main causes of catalysts deactivation, since the physicochemical properties of a nanoparticle may depend strongly on its size, shape and composition. For application as heterogeneous catalysts, the nanoparticle...... kinks and edges often play an important role for the catalytic activity. In order to preserve these sites, it is important to stabilize the supported nanoparticles with sizes of a few nanometers during operational conditions at often high temperatures and in the relevant gas environments. A prototypical...... nanocatalyst system for studying coarsening consists of Pt nanoparticles supported on an Al2O3 material which is relevant as an oxidation catalyst in diesel and lean-burn engine exhaust after-treatment technologies. In this study we address the effect on sintering of the shape of Pt nanoparticles supported...

  10. A study on synthesis and properties of Ag nanoparticles immobilized polyacrylamide hydrogel composites

    International Nuclear Information System (INIS)

    Saravanan, P.; Padmanabha Raju, M.; Alam, Sarfaraz

    2007-01-01

    Synthesis of Ag nanoparticles containing polyacrylamide (PAm) hydrogel composites was performed by free-radical cross-linking polymerization of acrylamide monomer in an aqueous medium containing Ag + ions. The Ag nanoparticle/PAm composites exhibit faint yellow colour and are found to stable under ambient conditions, without undergoing oxidation. TEM micrographs reveal the presence of nearly spherical and well-separated Ag nanoparticles with diameters in the range of 4-7 nm. UV-vis studies apparently show the characteristic surface plasmon band at ∼415 nm, for the existence of Ag nanoparticles within the hydrogel matrix. The effect of varying Ag + ion concentration within the PAm hydrogels on the amount of formation of Ag nanoparticles, as well as on the bulk properties of hydrogel nanocomposites such as equilibrium swelling, optical and electrical properties are studied. The Ag/PAm hydrogel nanocomposites have higher swelling ratio and lower electron transfer resistance than its corresponding conventional hydrogel

  11. Systematic study of ligand structures of metal oxide EUV nanoparticle photoresists

    KAUST Repository

    Jiang, Jing

    2015-03-19

    Ligand stabilized metal oxide nanoparticle resists are promising candidates for EUV lithography due to their high sensitivity for high-resolution patterning and high etching resistance. As ligand exchange is responsible for the patterning mechanism, we systematically studied the influence of ligand structures of metal oxide EUV nanoparticles on their sensitivity and dissolution behavior. ZrO2 nanoparticles were protected with various aromatic ligands with electron withdrawing and electron donating groups. These nanoparticles have lower sensitivity compared to those with aliphatic ligands suggesting the structures of these ligands is more important than their pka on resist sensitivity. The influence of ligand structure was further studied by comparing the nanoparticles’ solubility for a single type ligand to mixtures of ligands. The mixture of nanoparticles showed improved pattern quality. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

  12. In vitro transfection of bone marrow-derived dendritic cells with TATp-liposomes

    Directory of Open Access Journals (Sweden)

    Pappalardo JS

    2014-02-01

    Full Text Available Juan Sebastián Pappalardo,1–3 Cecilia A Langellotti,2 Sebastián Di Giacomo,1 Valeria Olivera,1 Valeria Quattrocchi,2 Patricia I Zamorano,1,2 William C Hartner,3 Tatyana S Levchenko,3 Vladimir P Torchilin3 1Virology Institute, Center for Research in Veterinary and Agronomic Sciences, National Institute for Agricultural Technology (INTA, Hurlingham, BA, Argentina; 2National Council for Scientific and Technical Research (CONICET, Autonomous City of Buenos Aires, Argentina; 3Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston, MA, USA Abstract: Dendritic cells (DC are antigen-presenting cells uniquely capable of priming naïve T cells and cross-presenting antigens, and they determine the type of immune response elicited against an antigen. TAT peptide (TATp, is an amphipathic, arginine-rich, cationic peptide that promotes penetration and translocation of various molecules and nanoparticles into cells. TATp-liposomes (TATp-L used for DC transfection were prepared using TATp derivatized with a lipid-terminated polymer capable of anchoring in the liposomal membrane. Here, we show that the addition of TATp to DNA-loaded liposomes increased the uptake of DNA in DC. DNA-loaded TATp-L increased the in vitro transfection efficiency in DC cultures as evidenced by a higher expression of the enhanced green fluorescent protein and bovine herpes virus type 1 glycoprotein D (gD. The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L, as indicated by the secretion of interleukin 6. Keywords: dendritic cell transfection, green fluorescent protein, bovine herpes virus 1 glycoprotein D, liposomes, TAT peptide, interleukin 6

  13. Freshwater ecotoxicity characterisation factor for metal oxide nanoparticles: A case study on titanium dioxide nanoparticle

    DEFF Research Database (Denmark)

    Salieri, Beatrice; Righi, Serena; Pasteris, Andrea

    2015-01-01

    when performing Life Cycle Impact Assessment, where characterization models and consequently characterization factors (CFs) for ENPs are missing. This paper aims to provide the freshwater ecotoxicity CF for titanium dioxide nanoparticles (nano-TiO2). The USEtox™ model has been selected...

  14. Comparative study of titania nanoparticles and nanotubes as antibacterial agents

    Science.gov (United States)

    Jing, Zhihong; Guo, Daojun; Wang, Weihua; Zhang, Shufang; Qi, Wei; Ling, Baoping

    2011-09-01

    Anatase titania nanoparticles with a high surface area (about 587.7 m 2/g) were synthesized by sol-gel method using isobutyl alcohol as solvent, and then anatase titania nanotubes with needlelike shape, which had diameters of about 5 nm and wall thickness of about 1 nm, could be obtained by microwave process using the above titania nanoparticles as precursors. Both titania nanoparticles and nanotubes were characterized through X-ray diffraction, transmission electron microscopy, high-resolution transmission electron microscopy, Fourier transform infrared spectroscopy, ultraviolet-visible spectroscopy, photoluminescence spectroscopy and nitrogen adsorption-desorption isotherm technique. The antibacterial activities of both titania nanoparticles and nanotubes against Escherichia coli ( E. coli) were developed by quantification and qualitative ways, e.g. microcalorimetric method and disk diffusion method. At the same time, their antibacterial activities against E. coli were also investigated in dark and under UV irradiation. As a result, both the titania nanoparticles and nanotubes had good antibacterial activities against E. coli due to their low inhibitory concentration and large diameter of antibacterial circle. In addition, the titania nanoparticles displayed higher antibacterial activities than those of the titania nanotubes under UV irradiation, though they presented similar antibacterial activities in dark. The differences in antibacterial activities between titania nanoparticles and nanotubes might be attributed to the changes of their microstructure in our works.

  15. Effects of parameters, plasmid dosages and topological structures on transfection efficiency of porcine fetal fibroblasts using different electroporators.

    Science.gov (United States)

    Zhong, Cui-Li; Li, Guo-Ling; Mo, Jian-Xin; Quan, Rong; Wang, Hao-Qiang; Li, Zi-Cong; Wu, Zhen-Fang; Zhang, Xian-Wei

    2017-10-20

    To obtain an ideal transfection efficiency of porcine fetal fibroblasts, fluorescence activated cell sorting (FACS) was used to optimize parameters for transfection of porcine fetal fibroblasts (PFFs) with ECM? 830, NEPA 21 and Nucleofector? 2b in different conditions such as electroporation parameters, plasmid dosages and topological structures. The results show that the optimum poring pulse parameter of NEPA 21 is voltage 200 V, continuous 3 ms, interval 50 ms, 3 times, voltage attenuation range of 10%; and the transfection efficiency of Nucleofector? 2b is highest under U-023 program. Under the optimum conditions, FACS analysis demonstrates that Nucleofector? 2b and ECM? 830 have the highest transfection efficiency when transfecting 10 μg supercoiled plasmids into PFFs, and 8 μg for NEPA 21. Supercoiled plasmids show higher transfection efficiencies than linearized plasmids. Moreover, Nucleofector? 2b has the highest transfection efficiency among the three electroporation instruments. This study paves the way to generate transgenic or gene editing pigs with high efficiency.

  16. Optimization of transfection parameters for ultrasound/SonoVue microbubble-mediated hAng-1 gene delivery in vitro.

    Science.gov (United States)

    Zhou, Qing; Chen, Jin-Ling; Chen, Qian; Wang, Xiao; Deng, Qing; Hu, Bo; Guo, Rui-Qiang

    2012-12-01

    This study aimed to explore the effects of microbubble concentration, gene dosage, cell-microbubble mixing mode and fetal bovine serum (FBS) on gene delivery. 293T cells were transfected with Sonovue microbubbles carrying the hAng-1 gene via ultrasound irradiation. Various ultrasound exposure parameters and microbubble and DNA concentrations were investigated. In addition, FBS and the cell suspension or adherent mode was explored. Transfection efficiency and cell viability were used to determine the optimal transfection parameters. hAng-1 gene transfection efficiency gradually increased with elongation of ultrasound exposure and increasing microbubble concentration. However, if ultrasound irradiation exceeded 1.5 W/cm² and 30 sec or the microbubble concentration was over 20%, hAng-1 gene expression was significantly decreased, coupled with extensive cell death. Gene transfection levels were low under DNA concentrations less than 15 µg/ml. Furthermore, the gene transfer rate was significantly increased under cell suspension mode; FBS had no effect on hAng-1 gene transfection. The integrity of hAng-1 DNA was not affected by ultrasonic irradiation under optimal conditions. The optimal transfection parameters for the hAng-1 gene and Sonovue microbubble were ultrasound exposure of 1.5 W/cm² and 30 sec, 20% microbubbles, 15 µg/ml of DNA and under cell suspension mode.

  17. Ecotoxicological studies of CdS nanoparticles on photosynthetic microorganisms.

    Science.gov (United States)

    Brayner, Roberta; Dahoumane, Si Amar; Nguyen, Julie Ngoc-Lan; Yéprémian, Claude; Djediat, Chakib; Couté, Alain; Fiévet, Fernand

    2011-03-01

    The potential ecotoxicity of nanosized cadmium sulfide (CdS), synthesized by the polyol process, was investigated using common Anabaena flos-aquae cyanobacteria and Euglena gracilis euglenoid microalgae. The photosynthetic activities of these microorganisms, after addition of free Cd2+ ions and CdS nanoparticles, varied with the presence of tri-n-octylphosphine oxide (TOPO) used to protect surface particle to avoid toxicity and also to control particle size and shape during the synthesis. The nanoparticle concentration was varied from 10(-3) to 5 x 10(-4) M. It was observed that the cadmium concentration, the addition of TOPO protective agent and the particle dissolution process in the culture medium play an important role during the ecotoxicological tests. Viability tests were followed by PAM fluorimetry. Cd2+ ions were very toxic for Anabaena flos aquae. The same behavior was observed after contact with CdS and CdS-TOPO nanoparticles. However, for Euglena gracilis, the photosynthetic activity was stable for more than 1 month in the presence of Cd2+ ions. Moreover, it was observed that the toxicity varies with the concentration of CdS and CdS-TOPO nanoparticles, both kind of nanoparticles are toxic for this microorganism. Transmission electron microscopy (TEM) analyses of microorganisms ultrathin sections showed that polysaccharides produced by Anabaena flos-aquae, after contact with CdS and CdS-TOPO nanoparticles, protect the microalgae against particle internalization. Only some particles were observed inside the cells. Moreover, the nanoparticle internalization was observed after contact with all nanoparticles in the presence of Euglena gracilis by endocytosis. All nanoparticles are inside vesicles formed by the cells.

  18. Enhanced transfection efficiency and reduced cytotoxicity of novel lipid-polymer hybrid nanoplexes

    DEFF Research Database (Denmark)

    Jain, Sanyog; Kumar, Sandeep; Agrawal, Ashish Kumar

    2013-01-01

    The present study reports the development, characterization, and evaluation of novel polyelectrolytes stabilized lipoplexes as a nonviral vector for gene delivery. In order to achieve the advantage of both DOTAP (1,2-dioleoyl-3-trimethylammonium propane) and PEI (high transfection efficiency...... size of 242.6 ± 9.4 nm and zeta potential of +23.1 ± 1.5 mV. Following development nanoplexes were evaluated for cellular uptake, nuclear colocalization, transfection efficiency, and cellular toxicity in MCF-7, HeLa, and HEK-293 cell lines. In support of our hypothesis nanoplexes exhibited higher...... uptake and nuclear colocalization in comparison with DOTAP/PC, DOTAP/DOPE lipoplexes, and PEI polyplexes. Nanoplexes also exhibited 50-80, 11-12, 6-7, and 5-6 fold higher transfection efficiency in comparison with DOTAP/PC-lipoplexes, DOTAP/DOPE-lipoplexes, PEI-polyplexes, and lipofectamine, respectively...

  19. Tetracycline-regulated transgene expression in hippocampal neurones following transfection with adenoviral vectors.

    Science.gov (United States)

    Harding, T C; Geddes, B J; Noel, J D; Murphy, D; Uney, J B

    1997-12-01

    A transfer system that enabled the efficient introduction of transgenes into neurones and the quantitative control of the expressed transgene would greatly facilitate studies into neuronal gene function. To develop such a system we incorporated the tetracycline (Tet)-responsive On/Off regulatory elements into type-5 adenoviral (Ad) vectors. Regulation of transgene expression following transfection was measured by placing the enhanced green fluorescent protein (EGFP) gene upstream of the Tet regulatory element. The results showed that cultures of primary hippocampal cells could be transfected with very high efficiency (<70%) by the AdTet-On and AdTet-Off systems. Following transfection with the AdTet-On system no EGFP-fluorescent cells could be detected until doxycycline was added. The AdTet-Off system showed the reverse transcriptional regulation, in that the addition of Tet caused EGFP fluorescence to be abolished.

  20. Study of synthesis and optical properties of Cu nanoparticles

    Science.gov (United States)

    Singh, Jaiveer; Devi Lodhi, Pavitra; Choudhary, K. K.; Kaurav, Netram

    2017-05-01

    Nanoparticles of Copper (Cu) have attracted great interest in recent years because of their unique physical and optical properties that are of industrial importance. To understand their basic properties, Cu nanoparticles were synthesized by Polyol method. The synthesized powder was characterized by X-ray diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). The average particle size and lattice parameter estimated by XRD were found to be ~42.5 nm and 3.617 Å respectively. The results suggest suitability of these nanoparticles as dopants in other materials such as polymer materials and oxides.

  1. Preparation and biological activity studies of resveratrol loaded ionically cross-linked chitosan-TPP nanoparticles.

    Science.gov (United States)

    Wu, Jie; Wang, Yaping; Yang, Hao; Liu, Xiangyu; Lu, Zhong

    2017-11-01

    Nanoparticles with size range of 10-500nm can be efficiently delivered into cancer cells by the Enhanced Permeability and Retention (EPR) effect. Here, we prepared resveratrol (Res) loaded chitosan (CS) nanoparticles with the size of 172-217nm by an ionic cross-linking method, with sodium tripolyphosphate (TPP) as the cross-linking agent, to improve the stability, solubility and tumors targeting of the natural anti-cancer drug Res. The prepared Res loaded CS-TPP nanoparticles presented long-term storage stability and UV light stability. The cumulative drug release from nanoparticles in mimetic tumor tissue condition (pH 6.5) was higher than that in physiological condition (pH 7.4). Further, Res-loaded CS-TPP nanoparticles maintained the antioxidant activity of Res even after UV light irradiation. Cell viability study shows that the as prepared drug loaded nanoparticles had similar antiproliferative activity on hepatocellular carcinoma cells SMMC 7721 and lower cytotoxicity on normal hepatocyte cells L02 compared with free Res. Fluorescence microscopy observation revealed that the nanoparticles were efficiently taken in by SMMC 7721 cells. This work indicates the potential use of drug loaded CS-TPP nanoparticles for the efficient delivery of bioactive Res for chemotherapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Induction heating studies of combustion synthesized MgFe2O4 nanoparticles for hyperthermia applications

    International Nuclear Information System (INIS)

    Khot, V.M.; Salunkhe, A.B.; Thorat, N.D.; Phadatare, M.R.; Pawar, S.H.

    2013-01-01

    The structural, magnetic and ac magnetically induced heating characteristics of combustion synthesized MgFe 2 O 4 nanoparticles have been investigated for application in magnetic particle hyperthermia. As prepared nanoparticles showed ferrimagnetic behavior at room temperature with magnetization of about 33.83 emu/g at ±15 kOe. The solid state MgFe 2 O 4 nanoparticles exhibited specific absorption rate (SAR) of about 297 W/g at physiological safe range of frequency and amplitude. The increase in SAR and heating temperature in ac magnetic field was thought to be due to enhancement in magnetic hysteresis loss caused by dipole–dipole interactions in combustion synthesized MgFe 2 O 4 nanoparticles. - Highlights: ► Highly crystalline pure MgFe 2 O 4 nanoparticles were synthesized by low temperature combustion. ► Effect of ac magnetic field and nanoparticles concentration on heating characteristics of MgFe 2 O 4 nanoparticles was studied. ► Combustion synthesized MgFe 2 O 4 nanoparticles show highest specific absorption rate of 297 Wg −1 . ► The reported high value of specific absorption rate is advantageous for its use in magnetic particle hyperthermia

  3. Nanoparticle-Mediated Mechanical Destruction of Cell Membranes: A Coarse-Grained Molecular Dynamics Study.

    Science.gov (United States)

    Zhang, Liuyang; Zhao, Yiping; Wang, Xianqiao

    2017-08-16

    The effects of binding mode, shape, binding strength, and rotational speed of actively rotating nanoparticles on the integrity of cell membranes have been systematically studied using dissipative particle dynamics simulations. With theoretical analyses of lipid density, surface tension, stress distribution, and water permeation, we demonstrate that the rotation of nanoparticles can provide a strong driving force for membrane rupture. The results show that nanoparticles embedded inside a cell membrane via endocytosis are more capable of producing large membrane deformations under active rotation than nanoparticles attached on the cell membrane surface. Nanoparticles with anisotropic shapes produce larger deformation and have a higher rupture efficiency than those with symmetric shapes. Our findings provide useful design guidelines for a general strategy based on utilizing mechanical forces to rupture cell membranes and therefore destroy the integrity of cells.

  4. A Study On Dispersion Stability Of Nickel Nanoparticles Synthesized By Wire Explosion In Liquid Media

    Directory of Open Access Journals (Sweden)

    Kim C.K.

    2015-06-01

    Full Text Available In this study, nickel nanoparticles were synthesized in ethanol using portable pulsed wire evaporation, which is a one-step physical method. From transmission electron microscopy images, it was found that the Ni nanoparticles exhibited a spherical shape with an average diameter of 7.3 nm. To prevent aggregation of the nickel nanoparticles, a polymer surfactant was added into the ethanol before the synthesis of nickel nanoparticles, and adsorbed on the freshly synthesized nickel nanoparticles during the wire explosion. The dispersion stability of the prepared nickel nanofluids was investigated by zeta-potential analyzer and Turbiscan optical analyzer. As a result, the optimum concentration of polymer surfactant to be added was suggested for the maximized dispersion stability of the nickel nanofluids.

  5. Density function theory study of the adsorption and dissociation of carbon monoxide on tungsten nanoparticles.

    Science.gov (United States)

    Weng, Meng-Hsiung; Ju, Shin-Pon; Chen, Hsin-Tsung; Chen, Hui-Lung; Lu, Jian-Ming; Lin, Ken-Huang; Lin, Jenn-Sen; Hsieh, Jin-Yuan; Yang, Hsi-Wen

    2013-02-01

    The adsorption and dissociation properties of carbon monoxide (CO) molecule on tungsten W(n) (n = 10-15) nanoparticles have been investigated by density-functional theory (DFT) calculations. The lowest-energy structures for W(n) (n = 10-15) nanoparticles are found by the basin-hopping method and big-bang method with the modified tight-binding many-body potential. We calculated the corresponding adsorption energies, C-O bond lengths and dissociation barriers for adsorption of CO on nanoparticles. The electronic properties of CO on nanoparticles are studied by the analysis of density of state and charge density. The characteristic of CO on W(n) nanoparticles are also compared with that of W bulk.

  6. Green synthesis of silver nanoparticles using Azadirachta indica leaf extract and its antimicrobial study

    Science.gov (United States)

    Roy, Pragyan; Das, Bhagyalaxmi; Mohanty, Abhipsa; Mohapatra, Sujata

    2017-11-01

    In this study, green synthesis of silver nanoparticles was done using leaf extracts of Azadirachta indica. The flavonoids and terpenoids present in the extract act as both reducing and capping agent. Microbes ( Escherichia coli and Gram-positive bacteria) were isolated from borewell water using selective media. The silver nanoparticles showed antimicrobial activities against Gram-positive bacteria and E. coli. However the silver nanoparticles were more effective against E. coli as compared to Gram-positive bacteria. Various techniques were used to characterize synthesized silver nanoparticles such as DLS and UV-visible spectrophotometer. The absorbance peak was in the range of 420-450 nm, that varied depending upon the variation in the concentration of neem extract. This is a very rapid and cost-effective method for generation of silver nanoparticle at room temperature, however, its exact dose in water purification has to be determined.

  7. Experimental study of mutagenous and mitosis modifying activity of silver nanoparticles

    Directory of Open Access Journals (Sweden)

    B. S. Kirbik

    2015-01-01

    Full Text Available Mutagenous and mitosis modifying impact of silver nanoparticles has been studied on outbred mice. Nanoparticles were of round shape with dimensions of 5-50 nm, size of generated organic shell of 2-5 nm, the quantity in 1 mcm3 makes 120-270. Metaphasic analysis of mice bone marrow cells was used as a testing technique. The frequency of chromosome aberrations and mitotic index of preparations were accounted. During single intraperitoneal administration of the agent in the dose of 250 mcg/kg the silver nanoparticles demonstrated mitosis stimulating activity. No mutagenous effect of silver nanoparticles by daily administration for 4 days of 25 mcg/kg and single administration in the dose of 250 mcg/kg has been registered, but there is statistically insignificant tendency of aberrant metaphases increase. Consequently silver nanoparticles in the investigated doses demonstrated no mutagenous activity and can be considered safe for mammalian cells.

  8. Modulation of synthetic parameters of cobalt nanoparticles: TEM, EDS, spectral and thermal studies

    Science.gov (United States)

    Chandra, Sulekh; Kumar, Avdhesh

    2012-12-01

    The study focuses on the modulation of synthetic parameters in order to influence the size, structure, composition and arrangement of nanoparticles of cobalt. Cobalt nanoparticles were synthesized by ethanolic solution of benzildiethylenetriamine in cobalt nitrate solution at 60 °C with stirring and refluxing leads to nanoparticles of cobalt. The morphology and structure of the synthesized nanoparticles of cobalt were characterized by Transmission Electron Microscopy (TEM), Powder X-ray Diffraction (XRD), Thermal Gravimetric Analysis (TGA), QELS Data and Infrared Spectroscopy (IR). Crystalline size was 20 nm determined from the sharp peak at 2θ = 25 °C from the powder XRD. TEM images of cobalt nanoparticles without reducing agent having the diameter 20 nm with spherical shape and black color.

  9. Cubic phase nanoparticles for sustained release of ibuprofen: formulation, characterization, and enhanced bioavailability study

    Science.gov (United States)

    Dian, Linghui; Yang, Zhiwen; Li, Feng; Wang, Zhouhua; Pan, Xin; Peng, Xinsheng; Huang, Xintian; Guo, Zhefei; Quan, Guilan; Shi, Xuan; Chen, Bao; Li, Ge; Wu, Chuanbin

    2013-01-01

    In order to improve the oral bioavailability of ibuprofen, ibuprofen-loaded cubic nanoparticles were prepared as a delivery system for aqueous formulations. The cubic inner structure was verified by cryogenic transmission electron microscopy. With an encapsulation efficiency greater than 85%, the ibuprofen-loaded cubic nanoparticles had a narrow size distribution around a mean size of 238 nm. Differential scanning calorimetry and X-ray diffraction determined that ibuprofen was in an amorphous and molecular form within the lipid matrix. The in vitro release of ibuprofen from cubic nanoparticles was greater than 80% at 24 hours, showing sustained characteristics. The pharmacokinetic study in beagle dogs showed improved absorption of ibuprofen from cubic nanoparticles compared to that of pure ibuprofen, with evidence of a longer half-life and a relative oral bioavailability of 222% (P ibuprofen-loaded cubic nanoparticles provide a promising carrier candidate with an efficient drug delivery for therapeutic treatment. PMID:23468008

  10. Induction of osteogenic differentiation of stem cells via a lyophilized microRNA reverse transfection formulation on a tissue culture plate

    Science.gov (United States)

    Wu, Kaimin; Xu, Jie; Liu, Mengyuan; Song, Wen; Yan, Jun; Gao, Shan; Zhao, Lingzhou; Zhang, Yumei

    2013-01-01

    MicroRNA (miRNA) regulation is a novel approach to manipulating the fate of mesenchymal stem cells, but an easy, safe, and highly efficient method of transfection is required. In this study, we developed an miRNA reverse transfection formulation by lyophilizing Lipofectamine 2000-miRNA lipoplexes on a tissue culture plate. The lipoplexes can be immobilized on a tissue culture plate with an intact pseudospherical structure and lyophilization without any lyoprotectant. In this study, reverse transfection resulted in highly efficient cellular uptake of miRNA and enabled significant manipulation of the intracellular target miRNA level. Reverse transfection formulations containing Lipofectamine 2000 1 μL per well generated much higher transfection efficiency without obvious cytotoxicity compared with conventional and other transfection methods. Further, the transfection efficiency of the reverse transfection formulations did not deteriorate during 90 days of storage at 4°C and −20°C. We then assessed the efficiency of the miRNA reverse transfection formulation in promoting osteogenic differentiation of mesenchymal stem cells. We found that transfection with anti-miR-138 and miR-148b was efficient for enhancing osteogenic differentiation, as indicated by enhanced osteogenesis-related gene expression, amount of alkaline phosphatase present, production of collagen, and matrix mineralization. Overall, the miRNA reverse transfection formulation developed in this study is a promising approach for miRNA transfection which can control stem cell fate and is suitable for loading miRNAs onto various biomaterials. PMID:23662054

  11. Transfection of Bacillus subtilis protoplasts by bacteriophage phi do7 DNA.

    OpenAIRE

    Perkins, J B; Dean, D H

    1983-01-01

    DNA from the Bacillus subtilis temperate bacteriophage phi do7 was found to efficiently transfect B. subtilis protoplasts; protoplast transfection was more efficient than competent cell transfection by a magnitude of 10(3). Unlike competent cell transfection, protoplast transfection did not require primary recombination, suggesting that phi do7 DNA enters the protoplast as double-stranded molecules.

  12. A statistical study on nanoparticle movements in a microfluidic channel.

    Science.gov (United States)

    Lee, Tae-Rin; Chang, Yoon-Suk; Choi, Jae-Boong; Kim, Young-Jin

    2011-01-01

    Microfluidic channels have received much attention because they can be used to control and transport nanoscale objects such as nanoparticles, nanowires, carbon nanotubes, DNA and cells. However, so far, practical channels have not been easy to design because they require very expensive fabrication and sensitive experiments. Numerical approaches can be alternatives or supplementary measures to predict the performance of new channels because they efficiently explain nanoscale multi-physics phenomena and successfully solve nanowire alignment and cell adhesion problems. In this paper, a newly updated immersed finite element method that accounts for collision force and Brownian motion as well as fluid-solid interaction is proposed, and is applied to simulate nanoparticle movements in a microfluidic channel. As part of the simulation, Brownian motion effects in a single nanoparticle focusing lens system are examined under different temperature conditions, and the resulting transport efficiencies are discussed. Furthermore, nanoparticle movements in a double focusing lens system are predicted to show the enhancement of focusing efficiency.

  13. The radio-sensitivity effect of E1A gene transfected by PEI on colon carcinoma cell in vitro

    International Nuclear Information System (INIS)

    Wu Yinxia; Liu Dongfang; Liu Yongbiao; Xu Dongmei; Yao Side; Sheng Kanglong

    2011-01-01

    As a neotype nonviral vector, (Polyethylenimine, PEI) has been studied in gene transfection experiment. This study was investigated the growth inhibition and radio-sensitizing effect of E1A gene transfected by PEI on human colon carcinoma cell in vitro. The PSV-E1A recombinant plasmid, which was designed for high-level expression of E1A gene in a variety of eukaryotic cell lines, was transfected into SW480 cells by PEI. The transfection was confirmed by RT-PCR and G418 was used to get colon carcinoma cells stably expressed E1A gene. The cell growth curve were investigated to observe the growth inhibition induced by E1A gene. The redistributions of cell cycle were analyzed by flow cytometry. Cells before and after transfection were treated with irradiation, then the changes of radiation-sensitivity were tested by MTT assay after 24 h meanwhile the expression of HER-2 gene in SW480 cells before and after transfection was detected by western-blot. As results, (1) the colon carcinoma cells expressed E1A gene was confirmed by G418. (2) The result of RT-PCR demonstrated that PEI could transfect plasmid psv-E1A and the cells could stably express E1A gene. (3) Flow cytometry revealed that E1A gene transfected into human colon carcinoma cell could induce S stage suppression (p<0.001) and G2/M stage arrest (p<0.001). (4) Compared with the Non-transfected cells, the E1A-transfected cells (SW480-E1A cells) grew slowly observed by MTT assay which was used to get the absorbance of SW480 cell and SW480-E1A cell. (5) The radiation-sensitivity of SW480 cells transfected with E1A gene was up-regulated obviously (p<0.001). (6) The E1A gene obviously down-regulated HER-2 protein expression in colon carcinoma cells. Anyway, PEI can transfect plasmid psv-E1A gene which can significantly inhibit the growth rate of SW480 cell. Moreover, it also obviously enhanced the cell sensitivity to irradiation. (authors)

  14. Quantum chemical studies of trace gas adsorption on ice nanoparticles

    Science.gov (United States)

    Schrems, Otto; Ignatov, Stanislav K.; Gadzhiev, Oleg B.; Masunov, Artem E.

    2013-04-01

    We have investigated the interaction of atmospheric trace gases with crystalline water ice particles of nanoscale size by modern quantum chemical methods. Small ice particles which can be formed in different altitudes play an important role in chemistry and physics of the Earth atmosphere. Knowledge about the uptake and incorporation of atmospheric trace gases in ice particles as well as their interactions with water molecules is very important for the understanding of processes at the air/ice interface. The interaction of the atmospheric trace gases with atmospheric ice nanoparticles is also an important issue for the development of modern physicochemical models. Usually, the interactions between trace gases and small particles considered theoretically apply small-size model complexes or the surface models representing only fragments of the ideal surface. Ice particles consisting of 48, 72, 216 and 270 water molecules with a distorted structure of hexagonal water ice Ih were studied using the new SCC-DFTBA method combining well the advantages of the DFT theory and semiempirical methods of quantum chemistry. The largest clusters correspond to the minimal nanoparticle size which are considered to be crystalline as determined experimentally. The clusters up to (H2O)72 were studied at the B3LYP/6-31++G(d,p) and B3LYP/6-311++G(2d,2p) levels. The larger clusters were studied using DFTBA and DFTB+ methods. Several adsorption complexes for the (H2O)270 water ice cluster were optimized at the RI-BLYP/6-31+G(d) theory level to verify the DFTB+ results. Trace gas molecules were coordinated on different sites of the nanoparticles corresponding to different ice Ih crystal planes: (0001), (10-10), (11-20). As atmospheric trace gases we have chosen CO, CO2, HCO*, HCOH*, HCHO, HCOOH and (HCO)2. which are the possible products and intermediates of the UV photolysis of organic molecules such as HCHCHO adsorbed on the ice surface. The structures of the corresponding coordination

  15. Noninvasive imaging of transplanted living functional cells transfected with a reporter estrogen receptor gene

    International Nuclear Information System (INIS)

    Takamatsu, Shinji; Furukawa, Takako; Mori, Tetsuya; Yonekura, Yoshiharu; Fujibayashi, Yasuhisa

    2005-01-01

    The transplantation of functional cells such as dopaminergic cells into damaged tissue is now clinically ongoing, but at present the population of surviving cells at the transplantation site mostly cannot be noninvasively examined. To visualize surviving transplanted functional cells using a noninvasive method, we chose the estrogen receptor ligand binding domain (ERL) as a reporter molecule and 16α-[ 18 F]-fluoro-17β-estradiol (FES) for its ligand. We used a mouse embryonic stem (ES) cell line for recipient cells as a model. To obtain ES cells that constitutively or inducibly express ERL, we transfected two types of expression vectors into EB5 parental ES cell line using the lipofection method and obtained about 30 clones for each of the two types of transfectants. Then, to examine the expression level of ERL, we performed Western blotting analysis. Ligand uptake experiments were carried out using [ 3 H]-estradiol with or without excessive unlabeled estradiol for control cells and ERL transfectants. Each selected clone was also used for in vivo positron emission tomography (PET) imaging studies involving FES in nude mice transplanted with control cells and ERL transfectants. In some of the clones transfected with the inducible-type ERL gene, protein was expressed much higher than in the controls. However, constitutive-type ERL gene-transfected ES cells showed no protein production in spite of their gene expression activity being considerably high. All clones also expressed equal levels of the Oct-3/4 gene, a marker of pluripotency, in comparison with the parental cells. Also, the specific uptake of [ 3 H]-estradiol was over 30 times higher in inducer-treated ERL-expressing ES cells compared to untreated control cells. Finally, by performing dynamic PET imaging, we successfully visualized ERL-expressing teratomas using FES

  16. Comparative Study on the Synergistic Action of Differentially Synthesized Silver Nanoparticles with β-Cephem Antibiotics and Chloramphenicol

    International Nuclear Information System (INIS)

    Hari, N.; Thomas, T.K.; Nair, A.J.

    2014-01-01

    Synergistic activity of cephem antibiotics with silver nanoparticles (Ag NPs) was investigated. Silver nanoparticles were synthesized through biological and chemical method. The combined action of β-lactam cephem antibiotics with both green and chemically synthesized silver nanoparticles enhances the antibacterial activity against wide range of antibiotic resistant pathogens and making them applicable to medical devices and microbial control systems. Synergistic activity of chloramphenicol with silver nanoparticles was also studied.

  17. Infectious alphavirus production from a simple plasmid transfection+

    Directory of Open Access Journals (Sweden)

    Olson Ken E

    2011-07-01

    Full Text Available Abstract We have developed a new method for producing infectious double subgenomic alphaviruses from plasmids transfected into mammalian cells. A double subgenomic Sindbis virus (TE3'2J was transcribed from a cytomegalovirus PolII promoter, which results in the production of infectious virus. Transfection of as little as 125 ng of plasmid is able to produce 1 × 108 plaque forming units/ml (PFU/ml of infectious virus 48 hours post-transfection. This system represents a more efficient method for producing recombinant Sindbis viruses.

  18. Enduring high-efficiency in vivo transfection of neurons with non-viral magnetoparticles in the rat visual cortex for optogenetic applications.

    Science.gov (United States)

    Soto-Sánchez, Cristina; Martínez-Navarrete, Gema; Humphreys, Lawrence; Puras, Gustavo; Zarate, Jon; Pedraz, José Luis; Fernández, Eduardo

    2015-05-01

    This work demonstrates the successful long-term transfection in vivo of a DNA plasmid vector in rat visual cortex neurons using the magnetofection technique. The transfection rates reached values of up to 97% of the neurons after 30days, comparable to those achieved by viral vectors. Immunohistochemical treatment with anti-EGFP antibodies enhanced the detection of the EYFP-channelrhodopsin expression throughout the dendritic trees and cell bodies. These results show that magnetic nanoparticles offer highly efficient and enduring in vivo high-rate transfection in identified neurons of an adult mammalian brain and suggest that the magnetotechnique facilitates the introduction of large functional genetic material like channelrhodopsin with safe non-viral vectors using minimally invasive approaches. Gene therapy may be one of the treatment modalities for neurological diseases in the future. The use of viral transfection remains a concern due to restrictions to the size limit of the genetic material able to be packed, as well as safety issues. In this work, the authors evaluated magnetoplexes as an alternative vehicle. The results showed very promising data in that these nanoparticles could offer high transfection efficiency. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Prenatal development toxicity study of zinc oxide nanoparticles in rats

    Directory of Open Access Journals (Sweden)

    Hong JS

    2014-12-01

    Full Text Available Jeong-Sup Hong,1,2 Myeong-Kyu Park,1 Min-Seok Kim,1 Jeong-Hyeon Lim,1 Gil-Jong Park,1 Eun-Ho Maeng,1 Jae-Ho Shin,3 Meyoung-Kon Kim,4 Jayoung Jeong,5 Jin-A Park,2 Jong-Choon Kim,6 Ho-Chul Shin2 1Health Care Research Laboratory, Korea Testing and Research Institute, Gimpo, South Korea; 2College of Veterinary Medicine, Konkuk University, Seoul, South Korea; 3Department of Biomedical Laboratory Science, Eulji University, Seongnam-si, South Korea; 4Department of Biochemistry and Molecular Biology, Korea University Medical School and College, Seoul, South Korea; 5Toxicological Research Division, National Institute of Food and Drug Safety Evaluation, Chungcheongbuk-do, South Korea; 6College of Veterinary Medicine, Chonnam National University, Gwangju, South Korea Abstract: This study investigated the potential adverse effects of zinc oxide nanoparticles ([ZnOSM20(+ NPs] zinc oxide nanoparticles, positively charged, 20 nm on pregnant dams and embryo–fetal development after maternal exposure over the period of gestational days 5–19 with Sprague-Dawley rats. ZnOSM20(+ NPs were administered to pregnant rats by gavage at 0, 100, 200, and 400 mg/kg/day. All dams were subjected to a cesarean section on gestational day 20, and all of the fetuses were examined for external, visceral, and skeletal alterations. Toxicity in the dams manifested as significantly decreased body weight after administration of 400 mg/kg/day NPs; reduced food consumption after administration of 200 and 400 mg/kg/day NPs; and decreased liver weight and increased adrenal glands weight after administration of 400 mg/kg/day NPs. However, no treatment-related difference in: number of corpora lutea; number of implantation sites; implantation rate (%; resorption; dead fetuses; litter size; fetal deaths and placental weights; and sex ratio were observed between the groups. On the other hand, significant decreases between treatment groups and controls were seen for fetal weights after

  20. Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells.

    Science.gov (United States)

    Palchetti, Sara; Pozzi, Daniela; Marchini, Cristina; Amici, Augusto; Andreani, Cristina; Bartolacci, Caterina; Digiacomo, Luca; Gambini, Valentina; Cardarelli, Francesco; Di Rienzo, Carmine; Peruzzi, Giovanna; Amenitsch, Heinz; Palermo, Rocco; Screpanti, Isabella; Caracciolo, Giulio

    2017-02-01

    To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine - the gold standard among transfection reagents - was used as a reference. On the basis of our results, we suggest that an additional transfection barrier impairs transfection efficiency, that is: low lipoplex concentration at the cell surface. Based on the acquired knowledge we propose an optimized transfection protocol that allowed us to efficiently transfect DND41, JURKAT, MOLT3, P12-ICHIKAWA, ALL-SILL, TALL-1 human T-cell acute lymphoblastic leukemia (T-ALL) cell lines known to be difficult-to-transfect by using non-viral vectors and where LFN-based technologies fail to give satisfactory results. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Studies on Optical and Electrical Properties of Hafnium Oxide Nanoparticles

    Science.gov (United States)

    Jayaraman, Venkatachalam; Sagadevan, Suresh; Sudhakar, Rajesh

    2017-07-01

    In this paper, the synthesis and physico-chemical properties of hafnium oxide nanoparticles (HfO2 NPs) are analyzed and reported. The synthesis was carried out by the precipitation route by using hafnium tetrachloride (HfCl4) as precursor material with potassium hydroxide (KOH) dissolved in Millipore water. In the precipitation technique, the chemical reaction is comparatively simple, low-cost and non-toxic compared to other synthetic methods. The synthesized HfO2 NPs were characterized by using powder x-ray diffraction (PXRD), ultraviolet-visible (UV-Vis) spectroscopy, Raman analysis, and high-resolution transmission electron microscopy (HRTEM). The monoclinic structure of the HfO2 NPs was resolved utilizing x-ray diffraction (XRD). The optical properties were studied from the UV-Vis absorption spectrum. The optical band gap of the HfO2NPs was observed to be 5.1 eV. The Raman spectrum shows the presence of HfO2 NPs. The HRTEM image showed that the HfO2 NPs were of spherical shape with an average particle size of around 28 nm. The energy-dispersive x-ray spectroscopy (EDS) spectrum obviously demonstrated the presence of HfO2 NPs. Analysis and studies on the dielectric properties of the HfO2 NPs such as the dielectric constant, the dielectric loss, and alternating current (AC) conductivity were carried out at varying frequencies and temperatures.

  2. Intracellular characterization of Gag VLP production by transient transfection of HEK 293 cells.

    Science.gov (United States)

    Cervera, Laura; González-Domínguez, Irene; Segura, María Mercedes; Gòdia, Francesc

    2017-11-01

    Transient transfection is a fast, flexible, and cost-effective approach to produce biological products. Despite the continued interest in transient transfection, little is known regarding the transfection process at the intracellular level, particularly for complex products, such as virus-like particles (VLPs). The kinetics of PEI-mediated transfection following an established in-house protocol is reported in this work with the aim of characterizing and understanding the complete process leading to VLP generation and identifying important events driving process improvement. For this purpose, DNA/PEI polyplexes' internalization in cells was tracked using Cy3 DNA staining. The production of a fluorescently labeled Gag polyprotein (a Gag-GFP fusion construct that forms fluorescent Gag-VLPs) was monitored by flow cytometry and confocal microscopy, and the VLP concentration in supernatants was measured by fluorometry. DNA/PEI polyplexes interact with the cell membrane immediately after polyplex addition to the cell culture. A linear increase in the number of cells expressing the protein is observed during the first 60 min of contact between the cells and polyplexes. No additional improvement in the number of cells expressing the protein (up to 60%) or VLP production (up to 1 × 10 10 VLPs/mL) is observed with additional contact time between the cells and polyplexes. Polyplexes can be detected in the cytoplasm of transfected cells as early as 1.5 h post-transfection (hpt) and reach the nucleus approximately 4 hpt. GFP fluorescence is observed homogeneously in the cytoplasm of transfected cells 24 hpt, but generalized VLP budding is not observed by microscopy until 48 hpt. Although all cells have internalized a polyplex soon after transfection, only a fraction of cells (60%) express the fluorescent Gag protein. VLP production kinetics was also studied. Fluorescence in the supernatant (enveloped VLPs) is 40% less than total fluorescence, supernatant plus pellet

  3. Characterization and carboplatin loaded chitosan nanoparticles for the chemotherapy against breast cancer in vitro studies.

    Science.gov (United States)

    Khan, Md Asad; Zafaryab, Md; Mehdi, Syed Hassan; Quadri, Javed; Rizvi, M Moshahid A

    2017-04-01

    Aim of the studies to synthesized chitosan nanoparticles by an ionic interaction procedure. The nanoparticles were characterized by physicochemical methods like, DLS, TEM, Surface potential measurements, FT-IR and DSC. The average particle size of chitosan and carboplatin nanoparticles was found to be 277.25±11.37nm and 289.30±8.15nm and zeta potential was found to be 31±3.14mV and 33±2.15mV respectively with low polydispersity index. The maximum entrapment of carboplatin in nanoparticles was a spherical shape with a positive charge. The maximum encapsulation and loading efficiencies of carboplatin (5mg/ml) were obtained to be 58.43% and 13.27% respectively. The nanocarboplatin was better blood compatibility as compared to chitosan nanoparticles. Finally, the cytotoxic effects of the carboplatin loaded chitosan nanoparticles were tested in-vitro against breast cancer (MCF-7) cell lines. Our studies showed that the chitosan nanoparticles could be used as a promising candidate for drug delivery for the therapeutic treatment of breast cancer. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Study of optimizing the process of Cadmium adsorption by synthesized silver nanoparticles using Chlorella vulgaris

    Directory of Open Access Journals (Sweden)

    Faezeh Sajadi

    2016-05-01

    Full Text Available Background and Aim: Cadmium (Cd is one of the most toxic heavy metals in water that mostly enters the water cycle through industrial waste water. Silver nanoparticles have the capacity to remove heavy metals from the water resources through the mechanism of adsorption. The present study aimed at producing  silver bio-nanoparticles and optimizing . Cd removal from aquatic solutions. Materials and Methods: Silver bio-nanoparticles were extracted via a micro-algae Chlorella vulgaris extract and silver nitrate synthesis. Then, the characteristics of the particles were  determined using FT-IR, XRD, SEM devices. In order to optimize Cadmium adsorption by means of silver nanoparticles, parameters including pH, reaction time, initial concentration of Cd and concentrations of nanoparticles were studied under different conditions. Results: The resulting nanoparticles were spherical, single and crystalline, whose sizes were 10-45 nm.  Under the condition of PH = 8, the initial concentration of cadmium 0.5 mg/L, adsorbent dosage of 0.5 mg, reaction time of 10 min, temperature of 300C and mixing speed of 200 rpm, 99% of cadmium was removed. Isotherm of Cadmium-ion adsorption followed Langmuir (R2> 0/96 (and Freundlich (R2> 0/94 models. Conclusion: Under optimal conditions, silver bio-nanoparticles had the capacity of quick and effective adsorption of cadmium. Thus, with a cheap, non-toxic and environmentally friendly method  can remove heavy metals in a short time.

  5. Effects of sodium lactate Ringer's injection on transfection of human protein kinase C-α antisense oligonucleotide in A549 lung cancer cells.

    Science.gov (United States)

    Wang, Z H; Sun, W W; Han, Y L; Ma, Z

    2016-08-26

    In the present study, we evaluated the effects of four solutions [Dulbecco's modified Eagle's medium (DMEM), sodium lactate Ringer's injection (SLRI), phosphate-buffered saline (PBS), and NaCl] on the transfection of the human protein kinase C-a antisense oligonucleotide (PKC-a ASO) aprinocarsen in human lung carcinoma A549 cells. Specifically, SLRI, DMEM, PBS, or NaCl were used as the growth solutions for A549 cells, and OPTI-MEM was used as the PKC-a ASO diluent for transfection. Additionally, SLRI, DMEM, PBS, or NaCl were used as both the growth solutions and diluents for transfection. The cell viability and transfection efficiency were determined. The results demonstrated that when SLRI was used as either the growth solution or both the growth solution and diluent for aprinocarsen transfection in A549 cells, the effects were close to the best effects observed with DMEM as the growth solution and OPTI-MEM as the diluent, which supported the transfection of aprinocarsen into the cells. Moreover, SLRI resulted in higher transfection efficiency than those of PBS and NaCl. In in vitro experiments, aprinocarsen effectively induced apoptosis in A549 cells. In conclusion, SLRI may replace PBS or NaCl in clinical trials as a transfection solution readily accepted by the human body. To our knowledge, this is the first report demonstrating the use of SLRI as a transfection solution in lung-cancer cell lines.

  6. Raman and fluorescence microscopy to study the internalization and dissolution of photosensitizer nanoparticles into living cells

    Science.gov (United States)

    Scalfi-Happ, Claudia; Steiner, Rudolf; Wittig, Rainer; Graefe, Susanna; Ryabova, Anastasia; Loschenov, Victor

    2015-07-01

    In this present study we applied Raman and fluorescence microscopy to investigate the internalisation, cellular distribution and effects on cell metabolism of photosensitizer nanoparticles for photodynamic therapy in fibroblasts and macrophages.

  7. Rapid pharmacokinetic and biodistribution studies using cholorotoxin-conjugated iron oxide nanoparticles: a novel non-radioactive method.

    Directory of Open Access Journals (Sweden)

    Michelle Jeung-Eun Lee

    2010-03-01

    Full Text Available Recent advances in nanotechnology have led to the development of biocompatible nanoparticles for in vivo molecular imaging and targeted therapy. Many nanoparticles have undesirable tissue distribution or unacceptably low serum half-lives. Pharmacokinetic (PK and biodistribution studies can help inform decisions determining particle size, coatings, or other features early in nanoparticle development. Unfortunately, these studies are rarely done in a timely fashion because many nanotechnology labs lack the resources and expertise to synthesize radioactive nanoparticles and evaluate them in mice.To address this problem, we developed an economical, radioactivity-free method for assessing serum half-life and tissue distribution of nanoparticles in mice. Iron oxide nanoparticles coated with chitosan and polyethylene glycol that utilize chlorotoxin as a targeting molecule have a serum half-life of 7-8 hours and the particles remain stable for extended periods of time in physiologic fluids and in vivo. Nanoparticles preferentially distribute to spleen and liver, presumably due to reticuloendothelial uptake. Other organs have very low levels of nanoparticles, which is ideal for imaging most cancers in the future. No acute toxicity was attributed to the nanoparticles.We report here a simple near-infrared fluorescence based methodology to assess PK properties of nanoparticles in order to integrate pharmacokinetic data into early nanoparticle design and synthesis. The nanoparticles tested demonstrate properties that are excellent for future clinical imaging strategies and potentially suitable for targeted therapy.

  8. Interactions of PLGA nanoparticles with blood components: protein adsorption, coagulation, activation of the complement system and hemolysis studies.

    Science.gov (United States)

    Fornaguera, Cristina; Calderó, Gabriela; Mitjans, Montserrat; Vinardell, Maria Pilar; Solans, Conxita; Vauthier, Christine

    2015-04-14

    The intravenous administration of poly(lactic-co-glycolic) acid (PLGA) nanoparticles has been widely reported as a promising alternative for delivery of drugs to specific cells. However, studies on their interaction with diverse blood components using different techniques are still lacking. Therefore, in the present work, the interaction of PLGA nanoparticles with blood components was described using different complementary techniques. The influence of different encapsulated compounds/functionalizing agents on these interactions was also reported. It is worth noting that all these techniques can be simply performed, without the need for highly sophisticated apparatus or skills. Moreover, their transference to industries and application of quality control could be easily performed. Serum albumin was adsorbed onto all types of tested nanoparticles. The saturation concentration was dependent on the nanoparticle size. In contrast, fibrinogen aggregation was dependent on nanoparticle surface charge. The complement activation was also influenced by the nanoparticle functionalization; the presence of a functionalizing agent increased complement activation, while the addition of an encapsulated compound only caused a slight increase. None of the nanoparticles influenced the coagulation cascade at low concentrations. However, at high concentrations, cationized nanoparticles did activate the coagulation cascade. Interactions of nanoparticles with erythrocytes did not reveal any hemolysis. Interactions of PLGA nanoparticles with blood proteins depended both on the nanoparticle properties and the protein studied. Independent of their loading/surface functionalization, PLGA nanoparticles did not influence the coagulation cascade and did not induce hemolysis of erythrocytes; they could be defined as safe concerning induction of embolization and cell lysis.

  9. DyNAvectors: dynamic constitutional vectors for adaptive DNA transfection.

    Science.gov (United States)

    Clima, Lilia; Peptanariu, Dragos; Pinteala, Mariana; Salic, Adrian; Barboiu, Mihail

    2015-12-25

    Dynamic constitutional frameworks, based on squalene, PEG and PEI components, reversibly connected to core centers, allow the efficient identification of adaptive vectors for good DNA transfection efficiency and are well tolerated by mammalian cells.

  10. C-terminal KDEL-modified cystatin C is retained in transfected CHO cells

    DEFF Research Database (Denmark)

    Johansen, Teit Eliot; Vogel, Charlotte Katrine; Schwartz, Thue W.

    1990-01-01

    The significance of a C-terminal tetrapeptide, Lys-Asp-Glu-Leu (KDEL), as a retention signal for the endoplasmatic reticulum was studied using cystatin C, a general thiol protease inhibitor, as the reporter protein. Clones of CHO cells were analyzed after stable transfection with eukaryotic...

  11. Fine structure study on low concentration zinc substituted hydroxyapatite nanoparticles

    International Nuclear Information System (INIS)

    Hu, Wei; Ma, Jun; Wang, Jianglin; Zhang, Shengmin

    2012-01-01

    The fine structure of zinc substituted hydroxyapatite was studied using experimental analysis and first-principles calculations. The synthetic hydroxyapatite nanoparticles containing low Zn concentration show rod-like morphology. The crystallite sizes and unit-cell volumes tended to decrease with the increased Zn concentration according to X-ray diffraction patterns. The Zn K-edge X-ray absorption spectra and fitting results suggest that the hydroxyapatite doped with 0.1 mole% zinc is different in the zinc coordination environments compared with that containing more zinc. The density function theory calculations were performed on zinc substituted hydroxyapatite. Two mechanisms included replacing calcium by zinc and inserting zinc along the hydroxyl column and were investigated, and the related substitution energies were calculated separately. It is found that the substitution energies are negative and lowest for inserting zinc between the two oxygen atoms along the hydroxyl column (c-axis). Combined with the spectral analysis, it is suggested that the inserting mechanism is favored for low concentration zinc substituted hydroxyapatite. Highlights: ► We investigate the fine structure of hydroxyapatite with low content of Zn. ► XANES spectra are similar but a little different at low zinc content. ► Zinc ions influence hydroxyapatite crystal formation and lattice parameters. ► Formation energies are calculated according to plane-wave density function theory. ► Low content of zinc prefers to locate at hydroxyl column in hydroxyapatite lattice.

  12. Proteomics study of silver nanoparticles toxicity on Oryza sativa L.

    Science.gov (United States)

    Mirzajani, Fateme; Askari, Hossein; Hamzelou, Sara; Schober, Yvonne; Römpp, Andreas; Ghassempour, Alireza; Spengler, Bernhard

    2014-10-01

    The increasing use of silver nanoparticles, (AgNPs), will inevitably result in their release into the environment and thereby cause the exposure to plants. It was claimed that using AgNPs is a safe and efficient method to preserve and treat agents of disease in agriculture. This study tries to understand the protein populations and sub-populations and follow up environmental AgNPs stresses. To accomplish these, the action of homemade spherical AgNPs colloidal suspension against Oryza sativa L. was investigated by a proteomic approach (2-DE and NanoLC/FT-ICR MS identification). Twenty-eight responsive (decrement/increment in abundance) proteins were identified. Proteomic results revealed that an exposure of O. sativa L., root with different concentrations of AgNPs resulted in an accumulation of protein precursors, indicative of the dissipation of a proton motive force. The identified proteins are involved in oxidative stress tolerance, Ca(2+) regulation and signaling, transcription and protein degradation, cell wall and DNA/RNA/protein direct damage, cell division and apoptosis. The expression pattern of these proteins and their possible involvement in the nontoxicity mechanisms were discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. A study of porosity of synthetic polymer nanoparticles using PALS

    Energy Technology Data Exchange (ETDEWEB)

    Pham, B; Smith, S V [Centre for Antimatter-Matter Studies, Australian Nuclear Science and Technology Organisation (ANSTO) NSW 2232 (Australia); Guagliardo, P; Williams, J; Samarin, S, E-mail: binh.pham@ansto.gov.au, E-mail: svs@ansto.gov.au [Centre for Antimatter-Matter Studies, School of Physics, University of Western Australia, WA 6009 (Australia)

    2011-01-01

    Positron annihilation lifetime spectroscopy (PALS) has been used to study the free volume in dry synthetic polymer nanoparticles of various sizes. A series of poly(styrene/divinyl benzene) particles with diameters in the range of 100 to 500 nm were synthesized and then carefully chemically treated using the sulfonation process, to increase their porosity. The particles were characterised by Scanning Electron Microscopy (SEM), light scattering and PALS. Light scattering gave larger size for the treated particles, reflecting the hydration effect and therefore the increase in porosity. PALS spectra of untreated and treated particles gave four and three life-time components, respectively. Analysis by PAScual version 1.3.0 program indicated there was a reduction in the intensity and the type of the micropores in the treated particles. The data suggest PALS is a sensitive tool for detecting changes in microporosity in particles. The conflicting results obtained for light scattering compared to PALS for chemically treated particles is difficult to resolve and suggests sample preparation of polymeric materials for PALS is the critical factor.

  14. A DFT study of Cu nanoparticles adsorbed on defective graphene

    Energy Technology Data Exchange (ETDEWEB)

    García-Rodríguez, D.E. [Universidad Politécnica de Aguascalientes, Calle Paseo San Gerardo No. 297 Fracc. San Gerardo, 20342 Aguascalientes, Ags. (Mexico); Mendoza-Huizar, L.H. [Universidad Autónoma del Estado de Hidalgo, Área Académica de Química, Ciudad del Conocimiento. Carretera Pachuca-Tulancigo Km. 4.5 Mineral de la Reforma, 42186 Hidalgo (Mexico); Díaz, C., E-mail: cristina.diaz@uam.es [Departamento de Química, Módulo 13, Universidad Autónoma de Madrid, 28049 Madrid (Spain); Condensed Matter Physics Center (IFIMAC), Universidad Autónoma de Madrid, 28049 Madrid (Spain); Institute for Advanced Research in Chemical Science (IAdChem), Universidad Autónoma de Madrid, 28049 Madrid (Spain)

    2017-08-01

    Highlights: • Cu{sub n} supported on graphene may be a promising electrode material for DBFC's cells. • Cu{sub n}/graphene interaction is rather local and size independent. • Cu{sub 13} anchors strongly to defects in graphene, while keeping its gas-phase properties. - Abstract: Metal nanoparticles adsorbed on graphene are systems of interest for processes relative to catalytic reactions and alternative energy production. Graphene decorated with Cu-nanoparticles, in particular, could be a good alternative material for electrodes in direct borohydride fuel cells. However our knowledge of this system is still very limited. Based on density functional theory, we have analyzed the interaction of Cu{sub n} nanoparticles (n = 4, 5, 6, 7, 13) with pristine and defective-graphene. We have considered two types of defects, a single vacancy (SV), and an extended lineal structural defect (ELSD), formed by heptagon-pentagon pairs. Our analysis has revealed the covalent character of the Cu{sub n}-graphene interaction for pristine- and ELSD-graphene, and a more ionic-like interaction for SV-graphene. Furthermore, our analysis shows that the interaction between the nanoparticles and the graphene is rather local, i.e., only the nanoparticle atoms close to the contact region are involved in the interaction, being the electronic contact region much higher for defective-graphene than for pristine-graphene. Thus, the higher the particle the lower its average electronic and structural distortion.

  15. A DFT study of Cu nanoparticles adsorbed on defective graphene

    International Nuclear Information System (INIS)

    García-Rodríguez, D.E.; Mendoza-Huizar, L.H.; Díaz, C.

    2017-01-01

    Highlights: • Cu n supported on graphene may be a promising electrode material for DBFC's cells. • Cu n /graphene interaction is rather local and size independent. • Cu 13 anchors strongly to defects in graphene, while keeping its gas-phase properties. - Abstract: Metal nanoparticles adsorbed on graphene are systems of interest for processes relative to catalytic reactions and alternative energy production. Graphene decorated with Cu-nanoparticles, in particular, could be a good alternative material for electrodes in direct borohydride fuel cells. However our knowledge of this system is still very limited. Based on density functional theory, we have analyzed the interaction of Cu n nanoparticles (n = 4, 5, 6, 7, 13) with pristine and defective-graphene. We have considered two types of defects, a single vacancy (SV), and an extended lineal structural defect (ELSD), formed by heptagon-pentagon pairs. Our analysis has revealed the covalent character of the Cu n -graphene interaction for pristine- and ELSD-graphene, and a more ionic-like interaction for SV-graphene. Furthermore, our analysis shows that the interaction between the nanoparticles and the graphene is rather local, i.e., only the nanoparticle atoms close to the contact region are involved in the interaction, being the electronic contact region much higher for defective-graphene than for pristine-graphene. Thus, the higher the particle the lower its average electronic and structural distortion.

  16. A study of the conjugation of CdSe nanoparticles with functional polyoxometalates involving aminoacids

    International Nuclear Information System (INIS)

    Gutul, T.

    2013-01-01

    CdSe nanoparticles (CdSe NPs) are regarded as nano markers and an important component for biomedical applications. In this study, CdSe NPs and polyoxometalates were synthesized; surface modification with 1-thioglycerol and (β-Ala) was carried out. Polyoxometalates, which cause an inhibitory effect on cancer cells, were conjugated to the nanoparticles. UV- VIS, IR, XRD, and TEM studies were performed to characterize the resulting CdSe NPs, polyoxometalates, and conjugates. (author)

  17. Nanoparticle-mediated photothermal therapy: a comparative study of heating for different particle types.

    Science.gov (United States)

    Pattani, Varun P; Tunnell, James W

    2012-10-01

    Near-infrared (NIR) absorbing plasmonic nanoparticles enhance photothermal therapy of tumors. In this procedure, systemically delivered gold nanoparticles preferentially accumulate at the tumor site and when irradiated using laser light, produce localized heat sufficient to damage tumor cells. Gold nanoshells and nanorods have been widely studied for this purpose, and while both exhibit strong NIR absorption, their overall absorption and scattering properties differ widely due to their geometry. In this paper, we compared the photothermal response of both nanoparticle types including the heat generation and photothermal efficiency. Tissue simulating phantoms, with varying concentrations of gold nanoparticles, were irradiated with a near-infrared diode laser while concurrently monitoring the surface temperature with an infrared camera. We calculated nanoshell and nanorod optical properties using the Mie solution and the discrete dipole approximation, respectively. In addition, we measured the heat generation of nanoshells and nanorods at the same optical density to determine the photothermal transduction efficiency for both nanoparticle types. We found that the gold nanoshells produced more heat than gold nanorods at equivalent number densities (# of nanoparticles/ml), whereas the nanorods generated more heat than nanoshells at equivalent extinction values at the irradiance wavelength. To reach an equivalent heat generation, we found that it was necessary to have ∼36× more nanorods than nanoshells. However, the gold nanorods were found to have two times the photothermal transduction efficiency than the gold nanoshells. For the nanoparticles tested, the nanoshells generated more heat, per nanoparticle, than nanorods, primarily due to their overall larger geometric cross-section. Conversely, we found that the gold nanorods had a higher photothermal efficiency than the gold nanoshells. In conclusion, the ideal choice of plasmonic nanoparticle requires not only per

  18. Study of dithiol monolayer as the interface for controlled deposition of gold nanoparticles

    International Nuclear Information System (INIS)

    Cichomski, M.; Tomaszewska, E.; Kosla, K.; Kozlowski, W.; Kowalczyk, P.J.; Grobelny, J.

    2011-01-01

    Self-assembled monolayer of dithiol molecules, deposited on polycrystalline Au (111), prepared at room atmosphere, was studied using scanning tunneling microscopy (STM) and X-ray photoelectron spectroscopy (XPS). Dithiols were used as interface, which chemically bonds to the deposited gold nanoparticles through strong covalent bonds. The size and size distribution of the deposited nanoparticles were measured using dynamic light scattering (DLS), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The AFM results showed that nanoparticles are immobilized and stable during scanning procedure and do not contaminate the AFM tip. The size of monodisperse nanoparticles obtained from the DLS measurements is slightly higher than that obtained from the AFM and SEM measurements. This is due to the fact that the DLS measures the hydrodynamic radius, dependent on the protective chemical layer on nanoparticles. - Research Highlights: → Dithiols molecules create chemically bounded layers on a Au (111) surface. → Gold nanoparticles can be chemically bounded to a self-assembled monolayer. → Nanoparticles are stable during AFM probe interactions.

  19. Synthesis, characterization and antibacterial study on the chitosan-functionalized Ag nanoparticles.

    Science.gov (United States)

    Biao, Linhai; Tan, Shengnan; Wang, Yuanlin; Guo, Ximin; Fu, Yujie; Xu, Fengjie; Zu, Yuangang; Liu, Zhiguo

    2017-07-01

    This study provided a facile, one-step hydrothermal method to synthesize stable Ag colloid in aqueous solution by utilizing chitosan as both reductant and stabilizer. The formation of chitosan-functionalized Ag nanoparticles was verified by UV-Vis, FTIR, TEM, AFM and XRD measurements. FTIR results revealed that the primary amine groups and amide groups of chitosan have specific interactions with the surface of Ag nanoparticles. The average diameter of the Ag nanoparticles is 10.0±5.4nm as determined by TEM. Ag nanoparticles are highly crystalline as revealed by HR-TEM and XRD measurements. The size and shape of Ag nanoparticles are also found to depend on the pH condition in the synthesis. Ag nanoparticles were the main products at pH5.0 whereas large Ag nanotriangle and truncated triangular nanoplate were dominant at pH4.0 in the synthesis. Due to its monodispersity and good stability, the chitosan-functionalized Ag colloid synthesized at pH5.0 was further tested for its antibacterial activities against gram-positive bacteria, gram-negative bacteria and fungus. The results of zone of inhibition, inhibition ratio and SEM characterization revealed that chitosan-functionalized Ag nanoparticles have great bactericidal efficiency against both bacteria and fungus. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Comparative studies on structural properties and antimicrobial potential of spinel ferrite nanoparticles synthesized using various methods

    Science.gov (United States)

    Baraliya, Jagdish D.; Rakhashiya, Purvi M.; Patel, Pooja P.; Thaker, Vrinda S.; Joshi, Hiren H.

    2017-05-01

    In this study, novel multifunctional magnetic iron-based nanoparticles (CoFe2O4) coated with silica, silica-DEG (diethylene glycol), PEG (polyethylene glycol) were synthesized using Auto Combustion Method (ACM), Co-precipitation Method (COPM), Citrate Precursor Method (CPM), Flash Combustion Method (FCM). These spinel ferrite nanoparticles also contain very high antibacterial properties to fulfill the requirements of a drug delivery system so that the antibiotic concentration could be minimized. A potential delivery system could be based on a ferromagnetic fluid. The effects of various preparation methods on the physical properties of the nanoparticles were examined. The nanoparticles were also tested against four human pathogenic bacteria (Gram negative E.coli, P. aeruginosa, Gram positive S. aureus, S. pyogenus) and two fungi (C. albicans, A.niger). It was revealed that a nanoparticle has strong antibacterial activity as compared to antifungal. Further, Gram positive bacteria are more affected than Gram negative bacteria. It was also clear that different methods of coating have great influence on the antimicrobial properties. It was observed that these nanoparticles have significantly different but potentially very high antimicrobial activities against the tested organisms than found elsewhere by other nanoparticles on the same organisms.

  1. Comparative Analysis of Non-viral Transfection Methods in Mouse Embryonic Fibroblast Cells.

    Science.gov (United States)

    Lee, Migi; Chea, Kathleen; Pyda, Rajyalakshmi; Chua, Melissa; Dominguez, Isabel

    2017-07-01

    Mouse embryonic fibroblast (MEF) cells are an important in vitro model for developmental biology, disease, and reprogramming studies. However, as with other primary cells, they are challenging to transfect. Although viral gene-delivery methods achieve high gene-delivery efficiency, challenges with cell mutagenesis and safety among others have led to the use and improvement of non-viral gene-delivery methods in MEF cells. Despite the importance of gene delivery in MEF cells, there is limited comparison of method/reagent efficacy. In this study, we compared the effectiveness of different gene-delivery methods and several reagents currently available in MEF cells by introducing a plasmid containing enhanced green fluorescent protein (EGFP). We analyze transfection efficiency by EGFP fluorescence. Our results suggest that two gene-delivery methods-electroporation and magnetofection in combination with a lipid reagent, are the most efficient transfection methods in MEF cells. This study provides a foundation for the selection of transfection methods or reagents when using MEF cells.

  2. Studying the mechanism of hybrid nanoparticle EUV photoresists

    KAUST Repository

    Zhang, Ben

    2015-03-23

    This work focuses on the investigation of dual tone patterning mechanism with hybrid inorganic/organic photoresists. Hafnium oxide (HfO2) modified with acrylic acid was prepared and the influence of electrolyte solutions as well as pH on its particle size change was investigated. The average particle size and zeta potential of the nanoparticles in different electrolyte solutions were measured. The results show that addition of different concentrations of electrolytes changed the hydrodynamic diameter of nanoparticles in water. Increased concentration of tetramethyl ammonium hydroxide (TMAH) caused the zeta potential of nanoparticles to change from positive to negative and its hydrodynamic diameter to increase from 40 nm to 165 nm. In addition, increasing concentration of triflic acid led to the decrease of particle size and zeta potential. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

  3. Sans Studies Insight Into Improving of Yield of Block Copolymer-Stabilized Gold Nanoparticles

    Science.gov (United States)

    Ray, Debes; Aswal, V. K.

    2010-01-01

    Triblock copolymer poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) are well known as dispersion stabilizers. It has also been recently found that they can act as reducing agents along with stabilizers and these two properties of block copolymers together have provided a single-step synthesis and stabilization of gold nanoparticles at ambient temperature. We have studied the synthesis of stable gold nanoparticle solutions using block copolymer P85. Gold nanoparticles are prepared from 1 wt% aqueous solution of P85 mixed with varying concentration of HAuCl4.3H2O salt in the range 0.001 to 0.1 wt%. Surface plasmon resonance (SPR) band in UV-visible absorption spectra confirm the formation of the gold nanoparticles and the maximum yield of the nanoparticles is found to be quite low at 0.005 wt% of the salt solution. Small-angle neutron scattering (SANS) measurements in these systems suggest that a very small fraction of the block copolymers (nanoparticles and remaining form their own micelles, which probably results in the low yield. This can be explained as on an average a high block copolymer-to-gold ion ratio r0 (22) is required for 1 wt% P85 in the reduction reaction to produce gold nanoparticles. Based on this understanding, a step-addition method is used to enhance the yield of gold nanoparticles by manifold where the gold salt is added in small steps to maintain higher value of r(>r0) and therefore continuous formation of nanoparticles.

  4. Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay.

    Science.gov (United States)

    Osorio, Johan S; Bionaz, Massimo

    2017-08-30

    Gene reporter technology (GRT) has opened several new avenues for monitoring biological events including the activation of transcription factors, which are central to the study of nutrigenomics. However, this technology relies heavily on the insertion of foreign plasmid DNA into the nuclei of cells (i.e., transfection), which can be very challenging and highly variable among cell types. The objective of this study was to investigate the optimal conditions to generate reliable GRT assay data on bovine immortalized cell lines, Madin Darby Bovine Kidney (MDBK) and bovine mammary epithelial alveolar (MACT) cells. Results are reported for two experiments. In Experiment 1, using 96 well-plate and a robotic inverted fluorescent microscope, we compared transfection efficiency among commercially available transfection reagents (TR) Lipofectamine® 3000 (Lipo3), Lipofectamine® LTX (LipoLTX), and TransIT-X2® (TransX2), three doses of TR (i.e., 0.15, 0.3, and 0.4μL/well), and three doses of Green Fluorescent Protein plasmid DNA (i.e., 10, 25, and 50ng/well). Transfection efficiency and mortality rate were analyzed using CellProfiler software. Transfection efficiency increased until the end of the experiment (20h post-transfection) at which point MACT had greater transfection than MDBK cells (16.3% vs. 2.2%). It is unclear the reason for the low transfection in MDBK cells. Maximal transfection efficiency was obtained with 0.3μL/well of LipoLTX plus 25ng/well of plasmid DNA (ca. 29.5±1.9%) and 0.15μL/well of LipoLTX plus 25ng/well of plasmid DNA (ca. 4.0±0.4%) for MACT and MDBK cells, respectively. The higher amount of TR and DNA was generally associated with higher cell mortality. Using high, medium, and low transfection efficiency conditions determined in Experiment 1, we performed a GRT assay for peroxisome proliferator-activated response element (PPRE) luciferase in MACT and MDBK cells treated with 10nM or 100nM of synthetic Peroxisome Proliferator-activated Receptor

  5. Fundamental studies of chalcogenide nanocrystals, carbonaceous nanoparticles, and chromatographic materials

    Science.gov (United States)

    Baker, Jared Scott

    2011-12-01

    The development of novel nanomaterials and the understanding of their fundamental physical and chemical properties represent an exciting area of research. These materials are continuously being sought for ever-increasing applications; finding their way into uses that influence mankind on a daily basis. Combining elements from traditional nanoparticle characterization with electrophoretic-based techniques, this dissertation presents the analysis of carbon nanoparticles (CNPs) generated from a novel source (candle soot) as well as a unique perspective on the reactivity and degradation process of magic-sized cadmium chalcogenide nanocrystals. One potential application of CNPs is their use as an alternative fluorophore in a separation-based sensor system. Laser-induced-fluorescence (LIF) is a commonly used manner of detection in this type of platform, but is limited in many cases by problems associated with the fluorophore. Carbon-based nanoparticles have the potential to improve upon traditional fluorophores in applications that make use of LIF as the detection scheme. CNPs were extracted from the carbonaceous material produced by the incomplete combustion of a candle. The soot was submitted to an oxidizing treatment and extraction/filtration procedures rendering watersoluble luminescent species. Electron microscopy was used to identify globular, amorphous structures in the nanometer size-range. An aqueous suspension of CNPs demonstrated excellent stability in terms of its electronic properties, showing little change in absorption and emission spectra upon storage under ambient conditions over a two-year period. Capitalizing on the strengths of capillary electrophoresis (CE) as a characterization technique, we have analyzed the negatively-charged CNPs in terms of charge and size by studying the influence of variable CE conditions on the resulting separation. Separations at different pH revealed a highly complex mixture of CNPs, containing species with large

  6. In vivo studies of transdermal nanoparticle delivery with microneedles using photoacoustic microscopy

    Science.gov (United States)

    Moothanchery, Mohesh; Seeni, Razina Z.; Xu, Chenjie; Pramanik, Manojit

    2017-01-01

    Microneedle technology allows micron-sized conduits to be formed within the outermost skin layers for both localized and systemic delivery of therapeutics including nanoparticles. Histological methods are often employed for characterization, and unfortunately do not allow for the in vivo visualization of the delivery process. This study presents the utilization of optical resolution-photoacoustic microscopy to characterize the transdermal delivery of nanoparticles using microneedles. Specifically, we observe the in vivo transdermal delivery of gold nanoparticles using microneedles in mice ear and study the penetration, diffusion, and spatial distribution of the nanoparticles in the tissue. The promising results reveal that photoacoustic microscopy can be used as a potential imaging modality for the in vivo characterization of microneedles based drug delivery. PMID:29296482

  7. Mimusops elengi bark extract mediated green synthesis of gold nanoparticles and study of its catalytic activity

    Science.gov (United States)

    Majumdar, Rakhi; Bag, Braja Gopal; Ghosh, Pooja

    2016-04-01

    The bark extract of Mimusops elengi is rich in different types of plant secondary metabolites such as flavonoids, tannins, triterpenoids and saponins. The present study shows the usefulness of the bark extract of Mimusops elengi for the green synthesis of gold nanoparticles in water at room temperature under very mild conditions. The synthesis of the gold nanoparticles was complete within a few minutes without any extra stabilizing or capping agents and the polyphenols present in the bark extract acted as both reducing as well as stabilizing agents. The synthesized colloidal gold nanoparticles were characterized by HRTEM, surface plasmon resonance spectroscopy and X-ray diffraction studies. The synthesized gold nanoparticles have been used as an efficient catalyst for the reduction of 3-nitrophenol and 4-nitrophenol to their corresponding aminophenols in water at room temperature.

  8. SANS study of Lysozyme vs. BSA protein adsorption on silica nanoparticles

    Science.gov (United States)

    Kumar, Sugam; Aswal, V. K.; Kohlbrecher, J.

    2012-06-01

    Lysozyme (M.W. 14.7 kD) and BSA (M.W. 66.7 kD) are two most commonly studied model proteins in literature. Lysozyme (cationic) and BSA (anionic) are oppositely charged at pH 7 and their interaction with anionic silica nanoparticles has been studied using small-angle neutron scattering (SANS). Measurements were carried out on fixed 1 wt% concentration of nanoparticles and varying concentration of protein in the range 0.5 to 2 wt%. It is found that both the proteins adsorb on the nanoparticles where strong interaction of lysozyme leads to the aggregation of nanoparticles but the system remains stable with BSA. Adsorption increases with protein concentration and has been found much larger for lysozyme.

  9. Structural studies of nanoparticles doped with rare-earth ions in oxyfluoride lead-silicate glasses

    Science.gov (United States)

    Kichanov, Sergey E.; Kozlenko, D. P.; Gorshkova, Yu. E.; Rachkovskaya, G. E.; Zakharevich, G. B.; Savenko, B. N.

    2018-03-01

    Formation of PbF2 nanoparticles as up-conversion luminescent centers in oxyfluoride lead-silicate glass matrixes doped with rare-earth oxides has been studied by means of small-angle neutron scattering and neutron diffraction methods. During a two-stage heat treatment process, amorphous nanoparticles with sizes of 5-7 nm were initially formed, then were aggregated into more complex clusters with average dimensions up to 30 nm. The thermal treating at higher temperature led to crystallization of amorphous nanoparticles and, as a result, the formation of up-conversion luminescent centers in studied lead-silicate glasses. The sizes of amorphous and crystalline nanoparticles, fractional dimensions of density fluctuations in glass materials, as well as the lattice parameters of a phase of PbF2 were calculated.

  10. Study of Coating Geometries and Photoluminescence Properties of Metal Nanoparticles/Graphite Composites

    Directory of Open Access Journals (Sweden)

    Pasquale Barone

    2014-01-01

    Full Text Available In this work we present the results of a study of growth and characterization of metal nanoparticles (Ag, Au, and Co/carbon surfaces. The nanoparticles grew by laser ablation technique and their dimensions were controlled by light scattering study and AFM microscopy before their insertion on graphite surface. Nanoparticles appear randomly disposed on carbon surfaces aggregating to form big particles only in the case of silver. The different behavior of metal nanoparticles on carbon surface was explained in terms of different metal wetting of surface, in agreement with previous theoretical results of He et al. Chemical information, obtained by X-ray photoelectron spectroscopy, indicated that the doping process is a simple physisorption while the interfacial interaction between particles and carbon layers causes local defects in graphite structure and the appearance of a strong photoluminescence signal for all composites. Moreover, the visible optical absorption decreases about 10% indicating the progressive metallization of carbon surface.

  11. Study of the Effect of Silver Nanoparticles on Extended Spectrum

    Directory of Open Access Journals (Sweden)

    Marzieh Karami

    2013-07-01

    Full Text Available Background and Objectives: Wide use of beta-lactam antibiotics increases bacterial resistance to these groups of antibiotics in pathogen bacteria through production of beta-lactamase enzyme. The present study was carried out with the aim of evaluating the isolation of ESBL producing Klebsiella spp. in clinical specimens and investigating the effect of silver nanoparticles on them. Methods: A total of 61 clinical Klebsiella isolates were examined in terms of production of extended spectrum beta-lactamase (ESBL through disk diffusion method using the antibiotics cefixime, ceftriaxone, ceftazidime, as well as beta-lactamase inhibitor clavulanic acid, and the antibiotics’ minimal inhibitory concentration (MIC values were determined by agar dilution method. Then, ESBL production was examined using standard ESBL disk for detection of deta-lactamase through DDT (double disk approximation test method. Subsequently, the effect of different concentrations of nanosilver solution on isolated bacteria was studied. Student t-test was used for analysis of the data.Results: Out of 61 multidrug resistant Klebsiella isolates, 51 (60.83% Klebsiella pneumonea and 10 (39.16% Klebsiella oxytoca were recognized. All samples were demonstrated to be positive in double disk method for proving ESBL production, and were sensitive to the nanosilver solution with a concentration of 500ppm.Conclusion: The obtained findings showed that increase in the concentration of nanosilver solutions had a direct correlation with inhibition zone diameter of ESBL producing Klebsiella spp in vitro and in completely aseptic condition. If it is proved that nanosilver solutions are non-toxic in vivo, they could be used as a new effective alternative to antibiotics.

  12. Construction of rat beta defensin-2 eukaryotic expression vector and expression in the transfected rat corneal epithelial cell

    Directory of Open Access Journals (Sweden)

    Jing Dan

    2017-03-01

    Full Text Available AIM: To construct a recombinant eukaryotic expression vector of rat beta defensin-2(rBD-2, transfect it into the rat corneal epithelial cells with lipofection, determine the expression of target gene in the transfected cells, and discuss the potentiality of recombinant plasmid expressed in corneal epithelial cells, hoping to provide an experimental foundation for further study on the antimicrobial activity of rBD-2 in vitro and in vivo and to assess the probability of defensins as a new application for infectious corneal diseases in the future. METHODS: The synthetic rBD-2 DNA fragment was inserted between the XhoI and BamHI restriction enzyme cutting sites of eukaryotic expression vector pIRES2-ZsGreen1 to construct the recombinant plasmid pIRES2-ZsGreen1-rBD-2, then transformed it into E.coli DH5α, positive clones were screened by kanamycin and identified with restriction endonucleases and sequencing analysis. Transfection into the rat corneal epithelial cells was performed by lipofection. Then the experiment was divided into three groups: rat corneal epithelial cell was transfected with the recombinant plasmid pIRES2- ZsGreen1-rBD-2, rat corneal epithelial cell was transfected with the empty plasmid pIRES2-ZsGreen1 and the non-transfected group. The inverted fluorescence microscope was used to observe the transfection process. At last, the level of rBD-2 mRNA expressed in the transfected cells and the control groups are compared by the real-time fluoresence relative quantitative PCR. RESULTS: The recombinant eukaryotic expression vector of pIRES2-ZsGreen1-rBD-2 was successfully constructed. The level of rBD-2 mRNA in transfected cells was significantly higher than that in control groups through the real-time fluorescence relative quantitative PCR. CONCLUSION: The recombinant eukaryotic expression vector pIRES2-ZsGreen1-rBD-2 could be transfected into rat corneal epithelial cells, and exogenous rBD-2 gene could be transcripted into mRNA in

  13. GENE TRANSFER ON Betta imbellis THROUGH TRANSFECTION METHOD WITH DIFFERENT DNA CONCENTRATION

    Directory of Open Access Journals (Sweden)

    Eni Kusrini

    2016-12-01

    Full Text Available Big size betta (Giant have a high economic value compared to normal size betta, and over expression of growth hormone gene can produce giant fish.  As an initial step of giant transgenic betta productions, this study was conducted in order to obtain DNA plasmid concentration which provide higher hatching and survival rate of betta larvae.  Construction of PhGH pCcBA gene contains growth hormone gene of Siamese catfish (PhGH and it is controlled by the CCBA promoter. Betta imbellis broodstocks were spawned naturally, and embryos were collected 1-2 minutes after spawning time. One hundred embryos were dipped in 2 mL of transfectan X-treme gene which containp CcBA-PhGH construction genes (50 µg/mL, on room temperature for about 30 minutes. Treatments on this study were different transfectant : DNA plasmid ratiosnamely:A (0,75 µL: 0,25 µL; B (0,75 µL : 0,50 µL; C (0,75 µL: 0,75 µL, D as Control 1(without transfectant, 0,25 µL DNA; E.as Control 2(0,75 µL transfectant, without DNA, and Fas control 3 (without transfectant and without DNA. Every treatments was repeated three times.  Transfection embryos were hatched on a container (1L Volume. Study results showed that hatching rate and larvae survival rate  (4 days after hatching on treatment A were the same with the control, but slightly higher than B and C treatments. PCR analysis with DNA template showing that PhGH gene were found on embryos and larvae (pooled sample of treatment A, B and C. Furthermore, RT-PCR analysis showing the existence of mRNA PhGH expression on embryos and larvae (pooled sample. Therefore, embryo transfection with transfectant ratio 0,75 µL and  DNA 0,25 µLshowing the best results.

  14. Efficient Transfection of siRNA by Peptide Dendrimer-Lipid Conjugates.

    Science.gov (United States)

    Kwok, Albert; Eggimann, Gabriela A; Heitz, Marc; Reymond, Jean-Louis; Hollfelder, Florian; Darbre, Tamis

    2016-12-02

    Efficient delivery of small interfering RNA (siRNA) into cells is the basis of target-gene-specific silencing and, ultimately, gene therapy. However, current transfection reagents are relatively inefficient, and very few studies provide the sort of systematic understanding based on structure-activity relationships that would provide rationales for their improvement. This work established peptide dendrimers (administered with cationic lipids) as siRNA transfection reagents and recorded structure-activity relationships that highlighted the importance of positive charge distribution in the two outer layers and a hydrophobic core as key features for efficient performance. These dendrimer-based transfection reagents work as well as highly optimised commercial reagents, yet show less toxicity and fewer off-target effects. Additionally, the degrees of freedom in the synthetic procedure will allow the placing of decisive recognition features to enhance and fine-tune transfection and cell specificity in the future. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Rigid aromatic linking moiety in cationic lipids for enhanced gene transfection efficiency.

    Science.gov (United States)

    Wang, Bing; Zhao, Rui-Mo; Zhang, Ji; Liu, Yan-Hong; Huang, Zheng; Yu, Qing-Ying; Yu, Xiao-Qi

    2017-08-18

    Although numerous cationic lipids have been developed as non-viral gene vectors, the structure-activity relationship (SAR) of these materials remains unclear and needs further investigation. In this work, a series of lysine-derived cationic lipids containing linkages with different rigidity were designed and synthesized. SAR studies showed that lipids with rigid aromatic linkage could promote the formation of tight liposomes and enhance DNA condensation, which is essential for the gene delivery process. These lipids could give much higher transfection efficiency than those containing more flexible aliphatic linkage in various cell lines. Moreover, the rigid aromatic linkage also affords the material higher serum tolerance ability. Flow cytometry assay revealed that the target lipids have good cellular uptake, while confocal microscopy observation showed weaker endosome escape than Lipofectamine 2000. To solve such problem and further increase the transfection efficiency, some lysosomotropic reagents were used to improve the endosome escape of lipoplex. As expected, higher transfection efficiency than Lipofectamine 2000 could be obtained via this strategy. Cytotoxicity assay showed that these lipids have lower toxicity in various cell lines than Lipofectamine 2000, suggesting their potential for further application. This work demonstrates that a rigid aromatic linkage might distinctly improve the gene transfection abilities of cationic lipids and affords information to construct safe and efficient gene vector towards practical application. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  16. Comparative study of synthesized silver and gold nanoparticles ...

    Indian Academy of Sciences (India)

    due to its distinctive properties like good conductivity, chem- ical stability and catalytic and antibacterial activities ... The cytotoxic and antioxidant activity exhibited by the leaves and the anti-hyperglycemic and antilipidemic ..... ver and gold nanoparticles using Morinda tinctoria fruit and. Couroupita guianensis flower extract, ...

  17. Dynamics of Magnetic Nanoparticles Studied by Neutron Scattering

    DEFF Research Database (Denmark)

    Hansen, Mikkel Fougt; Bødker, Franz; Mørup, Steen

    1997-01-01

    We present the first triple-axis neutron scattering measurements of magnetic fluctuations in nanoparticles using an antiferromagnetic reflection. Both the superparamagnetic relaxation and precession modes in similar to 15 nm hematite particles are: observed. The results have been consistently...... analyzed on the basis of a simple model with uniaxial anisotropy and the Neel-Brown theory for the relaxation....

  18. Comparative study of synthesized silver and gold nanoparticles ...

    Indian Academy of Sciences (India)

    in vitro anticancer efficacy of synthesized silver and gold nanoparticles using leaves extract of Bauhinia tomen- tosa Linn. ... AuNPs and aqueous extract of leaves confirmed by MTT assay exhibited IC50 concentrations of 28.125, 46.875 and. 50 μg ml ... applications such as the treatment of cancer, gene therapy and drug ...

  19. Comparative study of synthesized silver and gold nanoparticles ...

    Indian Academy of Sciences (India)

    Nanotechnology is an emerging field in science and technology, which can be applied to synthesize new materials at the nanoscale level. The present investigation aimed at comparing the synthesis, characterization andin vitro anticancer efficacy of synthesized silver and gold nanoparticles using leaves extract of Bauhinia ...

  20. Europium polyoxometalates encapsulated in silica nanoparticles - characterization and photoluminescence studies

    Energy Technology Data Exchange (ETDEWEB)

    Neves, Cristina S.; Granadeiro, Carlos M.; Cunha-Silva, Luis; Eaton, Peter; Balula, Salete S.; Pereira, Eulalia [REQUIMTE/Departamento de Quimica e Bioquimica, Faculdade de Ciencias, Universidade do Porto (Portugal); Ananias, Duarte [CICECO, Departamento de Quimica, Universidade de Aveiro (Portugal); Gago, Sandra [REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, Monte de Caparica (Portugal); Feio, Gabriel [CENIMAT/I3N, Departamento de Ciencia dos Materiais, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, Monte de Caparica (Portugal); Carvalho, Patricia A. [ICEMS/Departamento de Bioengenharia, Instituto Superior Tecnico, Lisboa (Portugal)

    2013-06-15

    The incorporation of europium polyoxometalates into silica nanoparticles can lead to a biocompatible nanomaterial with luminescent properties suitable for applications in biosensors, biological probes, and imaging. Keggin-type europium polyoxometalates Eu(PW{sub 11}){sub x} (x = 1 and 2) with different europium coordination environments were prepared by using simple methodologies and no expensive reactants. These luminescent compounds were then encapsulated into silica nanoparticles for the first time through the water-in-oil microemulsion methodology with a nonionic surfactant. The europium polyoxometalates and the nanoparticles were characterized by using several techniques [FTIR, FT-Raman, {sup 31}P magic angle spinning (MAS) NMR, and TEM/energy-dispersive X-ray spectroscopy (TEM-EDS), AFM, dynamic light scattering (DLS), and inductively coupled plasma MS (ICP-MS) analysis]. The stability of the material and the integrity of the europium compounds incorporated were also examined. Furthermore, the photoluminescence properties of the Eu(PW{sub 11}){sub x} rate at SiO{sub 2} nanomaterials were evaluated and compared with those of the free europium polyoxometalates. The silica surface of the most stable nanoparticles was successfully functionalized with appropriate organosilanes to enable the covalent binding of oligonucleotides. (Copyright copyright 2013 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  1. Structural and optical studies of pHEMA encapsulated ZnS:Ni2+ nanoparticles

    Science.gov (United States)

    Mohan, R.; Sankarrajan, S.; Thiruppathi, G.

    2015-07-01

    In this study, ZnS:Ni2+ nanostructures have been synthesized through chemical precipitation method using poly (2-hydroxyethyl methacrylate) (pHEMA) as capping agent. The structural, morphological and optical properties at different pHEMA concentration of ZnS:Ni2+ were studied by using X-ray diffraction (XRD), high resolution transmission electron microscopy (HR-TEM), UV-Vis Spectroscopy (UV-Vis), fourier transform infrared spectroscopy (FT-IR), Photoluminescence (PL). The Average crystalline size of the nanoparticles was found to be in the range of ∼3.59-4.36 nm. The surface morphological analysis reveals that the pHEMA capped nanoparticles showed homogeneous smooth surface. HR-TEM analysis reminds the original size of pHEMA capped nanoparticles. The band gap investigation revealed the size dependent of quantum confined nanoparticles. The immobilized nanoparticles in pHEMA matrix were verified by FT-IR studies. Novel luminescence properties have been observed for uncapped and pHEMA capped ZnS nanoparticles. The optimal capping concentration was successfully determined and its influence on photoluminescence behavior has been thoroughly analyzed.

  2. Morphology and thermal studies of zinc sulfide and cadmium sulfide nanoparticles in polyvinyl alcohol matrix

    Science.gov (United States)

    Osuntokun, Jejenija; Ajibade, Peter A.

    2016-09-01

    Zn(II) and Cd(II) metal complexes of 1-cyano-1-carboethoxyethylene-2,2-dithiolato-κS,S'-bis(N,N-dimethylthiourea-κS) have been synthesized and characterized with analytical and spectroscopic techniques. The complexes were thermolysed in hexadecylamine at 200 °C to prepare ZnS and CdS nanoparticles. The nanoparticles were characterized with scanning electron microscope (SEM), transmission electron microscope (TEM), and powder X-ray diffraction (p-XRD). TEM images showed spherically shaped nanoparticles, whose sizes are in the range 4.33-7.21 nm for ZnS and 4.95-7.7 nm CdS respectively and XRD confirmed cubic crystalline phases for the nanoparticles. The optical band gap energy evaluated from the absorption spectra are 2.88 eV (430 nm) and 2.81 eV (440 nm) for the ZnS and CdS nanoparticles respectively. The as-prepared metal sulfide nanoparticles were further incorporated into polyvinyl alcohol (PVA) to give ZnS/PVA and CdS/PVA composites. The polymer nanocomposites were studied to investigate their morphology and thermal properties relative to the pure PVA. XRD diffractions indicated that the crystalline phases of the nanoparticles and the sizes in PVA matrices remained unaltered. Infra-red spectra studies revealed interactions between the PVA and the metal sulfide nanoparticles and TGA studies show that the ZnS/PVA and CdS/PVA nanocomposites exhibit better thermal stability than the pure PVA.

  3. Morphology and thermal studies of zinc sulfide and cadmium sulfide nanoparticles in polyvinyl alcohol matrix

    Energy Technology Data Exchange (ETDEWEB)

    Osuntokun, Jejenija; Ajibade, Peter A., E-mail: pajibade@ufh.ac.za

    2016-09-01

    Zn(II) and Cd(II) metal complexes of 1-cyano-1-carboethoxyethylene-2,2-dithiolato–κS,S’–bis (N,N-dimethylthiourea–κS) have been synthesized and characterized with analytical and spectroscopic techniques. The complexes were thermolysed in hexadecylamine at 200 °C to prepare ZnS and CdS nanoparticles. The nanoparticles were characterized with scanning electron microscope (SEM), transmission electron microscope (TEM), and powder X-ray diffraction (p-XRD). TEM images showed spherically shaped nanoparticles, whose sizes are in the range 4.33–7.21 nm for ZnS and 4.95–7.7 nm CdS respectively and XRD confirmed cubic crystalline phases for the nanoparticles. The optical band gap energy evaluated from the absorption spectra are 2.88 eV (430 nm) and 2.81 eV (440 nm) for the ZnS and CdS nanoparticles respectively. The as-prepared metal sulfide nanoparticles were further incorporated into polyvinyl alcohol (PVA) to give ZnS/PVA and CdS/PVA composites. The polymer nanocomposites were studied to investigate their morphology and thermal properties relative to the pure PVA. XRD diffractions indicated that the crystalline phases of the nanoparticles and the sizes in PVA matrices remained unaltered. Infra-red spectra studies revealed interactions between the PVA and the metal sulfide nanoparticles and TGA studies show that the ZnS/PVA and CdS/PVA nanocomposites exhibit better thermal stability than the pure PVA.

  4. A multicenter study of using carbon nanoparticles to show sentinel lymph nodes in early gastric cancer.

    Science.gov (United States)

    Yan, Jun; Zheng, Xiaoling; Liu, Zhangyuanzhu; Yu, Jiang; Deng, Zhenwei; Xue, Fangqing; Zheng, Yu; Chen, Feng; Shi, Hong; Chen, Gang; Lu, Jianping; Cai, Lisheng; Cai, Mingzhi; Xiang, Gao; Hong, Yunfeng; Chen, Wenbo; Li, Guoxin

    2016-04-01

    Lymph node metastasis occurs in approximately 10% of early gastric cancer. Preoperative or intra-operative identification of lymph node metastasis in early gastric cancer is crucial for surgical planning. The purpose of this study was to evaluate the feasibility of using carbon nanoparticles to show sentinel lymph nodes (SLNs) in early gastric cancer. A multicenter study was performed between July 2012 and November 2014. Ninety-one patients with early gastric cancer identified by preoperative endoscopic ultrasonography were recruited. One milliliter carbon nanoparticles suspension, which is approved by Chinese Food and Drug Administration, was endoscopically injected into the submucosal layer at four points around the site of the primary tumor 6-12 h before surgery. Laparoscopic radical resection with D2 lymphadenectomy was performed. SLNs were defined as nodes that were black-dyed by carbon nanoparticles in greater omentum and lesser omentum near gastric cancer. Lymph node status and SLNs accuracy were confirmed by pathological analysis. All patients had black-dyed SLNs lying in greater omentum and/or lesser omentum. SLNs were easily found under laparoscopy. The mean number of SLNs was 4 (range 1-9). Carbon nanoparticles were around cancer in specimen. After pathological analysis, 10 patients (10.99%) had lymph node metastasis in 91 patients with early gastric cancer. SLNs were positive in 9 cases and negative in 82 cases. In pathology, carbon nanoparticles were seen in lymphatic vessels, lymphoid sinus, and macrophages in SLNs. When SLNs were positive, cancer cells were seen in lymph nodes. The sensitivity, specificity, and accuracy of black-dyed SLNs in early gastric cancers were 90, 100, and 98.9 %, respectively. No patient had any side effects of carbon nanoparticles in this study. It is feasible to use carbon nanoparticles to show SLNs in early gastric cancer. Carbon nanoparticles suspension is safe for submucosal injection.

  5. Liposome-based vascular endothelial growth factor-165 transfection with skeletal myoblast for treatment of ischaemic limb disease.

    Science.gov (United States)

    Ye, Lei; Haider, Husnain Kh; Esa, Wahidah Bte; Su, Liping; Law, Peter K; Zhang, Wei; Lim, Yeanteng; Poh, Kian Keong; Sim, Eugene K W

    2010-01-01

    The study aims to use cholesterol (Chol) + DOTAP liposome (CD liposome) based human vascular endothelial growth factor-165 (VEGF(165)) gene transfer into skeletal myoblasts (SkMs) for treatment of acute hind limb ischaemia in a rabbit model. The feasibility and efficacy of CD liposome mediated gene transfer with rabbit SkMs were characterized using plasmid carrying enhanced green fluorescent protein (pEGFP) and assessed by flow cytometry. After optimization, SkMs were transfected with CD lipoplexes carrying plasmid-VEGF(165) (CD-pVEGF(165)) and transplanted into rabbit ischaemic limb. Animals were randomized to receive intramuscular injection of Medium199 (M199; group 1), non-transfected SkM (group 2) or CD-pVEGF(165) transfected SkM (group 3). Flow cytometry revealed that up to 16% rabbit SkMs were successfully transfected with pEGFP. Based on the optimized transfection condition, transfected rabbit SkM expressed VEGF(165) up to day 18 with peak at day 2. SkMs were observed in all cell-transplanted groups, as visualized with 6-diamidino-2-phenylindole and bromodeoxyuridine. Angiographic blood vessel score revealed increased collateral vessel development in group 3 (39.7 +/- 2.0) compared with group 2 (21.6 +/- 1.1%, P limb and may serve as a safe and new therapeutic modality for the repair of acute ischaemic limb disease.

  6. Oral insulin delivery by self-assembled chitosan nanoparticles: In vitro and in vivo studies in diabetic animal model

    Energy Technology Data Exchange (ETDEWEB)

    Mukhopadhyay, Piyasi; Sarkar, Kishor [Department of Polymer Science and Technology, 92, A.P.C. Road, Kolkata-700009, University of Calcutta (India); Chakraborty, Mousumi; Bhattacharya, Sourav; Mishra, Roshnara [Department of Physiology, 92, A.P.C. Road, Kolkata-700009, University of Calcutta (India); Kundu, P.P., E-mail: ppk923@yahoo.com [Department of Polymer Science and Technology, 92, A.P.C. Road, Kolkata-700009, University of Calcutta (India)

    2013-01-01

    We have developed self-assembled chitosan/insulin nanoparticles for successful oral insulin delivery. The main purpose of our study is to prepare chitosan/insulin nanoparticles by self-assembly method, to characterize them and to evaluate their efficiency in vivo diabetic model. The size and morphology of the nanoparticles were analyzed by dynamic light scattering (DLS), atomic force microscopy (AFM) and scanning electron microscopy (SEM). The average particle size ranged from 200 to 550 nm, with almost spherical or sub spherical shape. An average insulin encapsulation within the nanoparticles was {approx} 85%. In vitro release study showed that the nanoparticles were also efficient in retaining good amount of insulin in simulated gastric condition, while significant amount of insulin release was noticed in simulated intestinal condition. The oral administrations of chitosan/insulin nanoparticles were effective in lowering the blood glucose level of alloxan-induced diabetic mice. Thus, self-assembled chitosan/insulin nanoparticles show promising effects as potential insulin carrier system in animal models. Highlights: Black-Right-Pointing-Pointer Self-assembled chitosan/insulin nanoparticle preparation. Black-Right-Pointing-Pointer Almost spherical or sub-spherical nanoparticles observed under microscope. Black-Right-Pointing-Pointer Good insulin encapsulation of the nanoparticles. Black-Right-Pointing-Pointer Nanoparticles reduced blood glucose level significantly in diabetic mice.

  7. A study of the properties of core/shell/shell Ag/FeCo/Ag nanoparticles

    Science.gov (United States)

    Kamzin, A. S.; Takahashi, M.; Maenosono, S.; Valiullin, A. A.

    2017-10-01

    The properties of heterophase core/shell/shell Ag/FeCo/Ag nanoparticles synthesized via a plasma method that are promising for biological applications are studied. As is established, the core/shell/shell Ag/FeCo/Ag nanoparticles exhibit a superparamagnetic state at room temperature that allows one to manage the hyperthermia process. The magnetic characteristics of core/shell/shell Ag/FeCo/Ag nanoparticles are interpreted by assuming partial oxidation of the surface layer of a ferromagnetic FeCo shell and formation of the antiferromagnetic CoxFe1-xO layer on the FeCo surface. The interaction between the surface antiferromagnetic CoxFe1-xO layer and the ferromagnetic FeCo shell causes the emergence of the exchange bias in Ag/FeCo/Ag nanoparticles.

  8. Micro-Raman scattering studies of Ge-Sb-Te bulk crystals and nanoparticles

    International Nuclear Information System (INIS)

    Cho, E.; Yoon, S.; Yoon, H. R.; Jo, W.

    2006-01-01

    We measured micro-Raman scattering spectra of commercially available Ge-Sb-Te (GST) bulk crystals and GST nanoparticles which were synthesized using a pulsed laser ablation method. The lack of the amorphous Te-Te stretching mode near 150 cm -1 from the Raman spectrum of the bulk sample indicated that the sample was well-crystallized. We also measured GST nanoparticles with different growth conditions, from which we could get information towards the optimal growth conditions for better crystallinity of the GST nanoparticles. Our results suggest that through local structural information, micro-Raman scattering spectroscopy can be used to study the phases and the phase changes in the GST bulk crystals and nanoparticles which is being developed for low-power non-volatile memory applications.

  9. Size-dependent interaction of gold nanoparticles with transport protein: A spectroscopic study

    International Nuclear Information System (INIS)

    Pramanik, Smritimoy; Banerjee, Paltu; Sarkar, Arindam; Bhattacharya, Subhash Chandra

    2008-01-01

    Gold nanoparticles of different sizes have been synthesized using sodium citrate as a reducing agent for tetrachloroauric (III) acid. The formed gold nanoparticles have been characterized by the UV-visible and transmission electron microscopy (TEM) measurements. The different sized gold nanoparticles have been used to study the interaction with model transport protein, bovine serum albumin (BSA). Experimental results reveal that BSA molecules adsorbed on the metallic surfaces, suffer strong quenching of their fluorescence and the rate of quenching efficiency is different for different particle size. The analysis of the quenching results has been performed in terms of the Stern-Volmer equation. The mechanism of quenching of fluorescence has been explained. The extent of adsorption of BSA on the gold nanoparticles has been estimated

  10. Spectroscopic studies of energy transfer in fluorene co-polymer blend nanoparticles

    Science.gov (United States)

    Gao, Jian; Grey, John K.

    2012-01-01

    Nanoparticles of poly(9,9-dioctylfluorene-co-bis-N,N-(4-butylphenyl)-bis-N,N-phenyl-1,4-phenylenediamine) [PFB] and poly(9,9-dioctylfluorene-co-benzothiadiazole) [F8BT] (1:1 w/w) were studied using scanned probe and single particle spectroscopy techniques. Photoluminescence (PL spectra of ∼58 and ∼100 nm PFB/F8BT nanoparticles show efficient energy transfer from the PFB (donor) component to the F8BT (acceptor) component that is independent of particle size. We propose that nanoparticles are phase segregated into discrete PFB/F8BT nanodomains on the order of ∼20-40 nm for both particle sizes. Pressure-dependent nanoparticle PL spectra support this assignment where lineshape maxima of each component red-shift in a similar manner due to increased interchain packing within the single nanodomains.

  11. SANS study of interaction of silica nanoparticles with BSA protein and their resultant structure

    Science.gov (United States)

    Yadav, Indresh; Aswal, V. K.; Kohlbrecher, J.

    2014-04-01

    Small angle neutron scattering (SANS) has been carried out to study the interaction of anionic silica nanoparticles (88 Å) with globular protein Bovine Serum Albumin (BSA) (M.W. 66.4 kD) in aqueous solution. The measurements have been carried out on fixed concentration (1 wt %) of Ludox silica nanoparticles with varying concentration of BSA (0-5 wt %) at pH7. Results show that silica nanoparticles and BSA coexist as individual entities at low concentration of BSA where electrostatic repulsive interactions between them prevent their aggregation. However, as the concentration of BSA increases (≥ 0.5 wt %), it induces the attractive depletion interaction among nanoparticles leading to finally their aggregation at higher BSA concentration (2 wt %). The aggregates are found to be governed by the diffusion limited aggregation (DLA) morphology of fractal nature having fractal dimension about 2.4.

  12. SANS study of interaction of silica nanoparticles with BSA protein and their resultant structure

    Energy Technology Data Exchange (ETDEWEB)

    Yadav, Indresh, E-mail: vkaswal@barc.gov.in; Aswal, V. K., E-mail: vkaswal@barc.gov.in [Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai-400085 (India); Kohlbrecher, J. [Laboratory for Neutron Scattering, Paul Scherrer Institute, CH-5232 PSI Villigen Switzerland (Switzerland)

    2014-04-24

    Small angle neutron scattering (SANS) has been carried out to study the interaction of anionic silica nanoparticles (88 Å) with globular protein Bovine Serum Albumin (BSA) (M.W. 66.4 kD) in aqueous solution. The measurements have been carried out on fixed concentration (1 wt %) of Ludox silica nanoparticles with varying concentration of BSA (0–5 wt %) at pH7. Results show that silica nanoparticles and BSA coexist as individual entities at low concentration of BSA where electrostatic repulsive interactions between them prevent their aggregation. However, as the concentration of BSA increases (≥ 0.5 wt %), it induces the attractive depletion interaction among nanoparticles leading to finally their aggregation at higher BSA concentration (2 wt %). The aggregates are found to be governed by the diffusion limited aggregation (DLA) morphology of fractal nature having fractal dimension about 2.4.

  13. Toxicity of Transition Metal Oxide Nanoparticles: Recent Insights from in vitro Studies

    Directory of Open Access Journals (Sweden)

    Robert S. Aronstam

    2010-10-01

    Full Text Available Nanotechnology has evolved to play a prominent role in our economy. Increased use of nanomaterials poses potential human health risk. It is therefore critical to understand the nature and origin of the toxicity imposed by nanomaterials (nanotoxicity. In this article we review the toxicity of the transition metal oxides in the 4th period that are widely used in industry and biotechnology. Nanoparticle toxicity is compellingly related to oxidative stress and alteration of calcium homeostasis, gene expression, pro-inflammatory responses, and cellular signaling events. The precise physicochemical properties that dictate the toxicity of nanoparticles have yet to be defined, but may include element-specific surface catalytic activity (e.g., metallic, semiconducting properties, nanoparticle uptake, or nanoparticle dissolution. These in vitro studies substantially advance our understanding in mechanisms of toxicity, which may lead to safer design of nanomaterials.

  14. SANS study of interaction of silica nanoparticles with BSA protein and their resultant structure

    International Nuclear Information System (INIS)

    Yadav, Indresh; Aswal, V. K.; Kohlbrecher, J.

    2014-01-01

    Small angle neutron scattering (SANS) has been carried out to study the interaction of anionic silica nanoparticles (88 Å) with globular protein Bovine Serum Albumin (BSA) (M.W. 66.4 kD) in aqueous solution. The measurements have been carried out on fixed concentration (1 wt %) of Ludox silica nanoparticles with varying concentration of BSA (0–5 wt %) at pH7. Results show that silica nanoparticles and BSA coexist as individual entities at low concentration of BSA where electrostatic repulsive interactions between them prevent their aggregation. However, as the concentration of BSA increases (≥ 0.5 wt %), it induces the attractive depletion interaction among nanoparticles leading to finally their aggregation at higher BSA concentration (2 wt %). The aggregates are found to be governed by the diffusion limited aggregation (DLA) morphology of fractal nature having fractal dimension about 2.4

  15. Study of Iron oxide nanoparticles using Mössbauer spectroscopy with a high velocity resolution

    Science.gov (United States)

    Oshtrakh, M. I.; Ushakov, M. V.; Šepelák, V.; Semionkin, V. A.; Morais, P. C.

    2016-01-01

    Iron oxide (magnetite and maghemite) nanoparticles developed for magnetic fluids were studied using Mössbauer spectroscopy with a high velocity resolution at 295 and 90 K. The recorded Mössbauer spectra have demonstrated that usual physical models based on octahedral and tetrahedral sites were not suitable for fitting. Alternatively, the Mössbauer spectra were nicely fitted using a large number of magnetic sextets. The obtained results showed that the Mössbauer spectra and the assessed parameters were different for nanoparticles as-prepared and dispersed in the dispersing fluid at 295 K. We claim that this finding is mainly due to the interaction of polar molecules with Iron cations at nanoparticle's surface or due to the surface coating using carboxylic-terminated molecules. It is assumed that the large number of spectral components may be related to complexity of the nanoparticle's characteristics and deviations from stoichiometry, including in the latter the influence of the oxidation of magnetite towards maghemite.

  16. Study of mechanism of enhanced antibacterial activity by green synthesis of silver nanoparticles

    Science.gov (United States)

    Parashar, Upendra Kumar; Kumar, Vinod; Bera, Tanmay; Saxena, Preeti S.; Nath, Gopal; Srivastava, Sunil K.; Giri, Rajiv; Srivastava, Anchal

    2011-10-01

    The extensive use of silver nanoparticles needs a synthesis process that is greener without compromising their properties. The present study describes a novel green synthesis of silver nanoparticles using Guava (Psidium guajava) leaf extract. In order to compare with the conventionally synthesized ones, we also prepared Ag-NPs by chemical reduction. Their optical and morphological characteristics were thoroughly investigated and tested for their antibacterial properties on Escherichia coli. The green synthesized silver nanoparticles showed better antibacterial properties than their chemical counterparts even though there was not much difference between their morphologies. Fourier transform infrared (FTIR) spectroscopic analysis of the used extract and as-synthesized silver nanoparticles suggests the possible reduction of Ag + by the water-soluble ingredients of the guava leaf like tannins, eugenol and flavonoids. The possible reaction mechanism for the reduction of Ag + has been proposed and discussed. The time-dependent electron micrographs and the simulation studies indicated that a physical interaction between the silver nanoparticles and the bacterial cell membrane may be responsible for this effect. Based on the findings, it seems very reasonable to believe that this greener way of synthesizing silver nanoparticles is not just an environmentally viable technique but it also opens up scope to improve their antibacterial properties.

  17. Study of mechanism of enhanced antibacterial activity by green synthesis of silver nanoparticles

    International Nuclear Information System (INIS)

    Parashar, Upendra Kumar; Srivastava, Sunil K; Srivastava, Anchal; Kumar, Vinod; Saxena, Preeti S; Bera, Tanmay; Nath, Gopal; Giri, Rajiv

    2011-01-01

    The extensive use of silver nanoparticles needs a synthesis process that is greener without compromising their properties. The present study describes a novel green synthesis of silver nanoparticles using Guava (Psidium guajava) leaf extract. In order to compare with the conventionally synthesized ones, we also prepared Ag-NPs by chemical reduction. Their optical and morphological characteristics were thoroughly investigated and tested for their antibacterial properties on Escherichia coli. The green synthesized silver nanoparticles showed better antibacterial properties than their chemical counterparts even though there was not much difference between their morphologies. Fourier transform infrared (FTIR) spectroscopic analysis of the used extract and as-synthesized silver nanoparticles suggests the possible reduction of Ag + by the water-soluble ingredients of the guava leaf like tannins, eugenol and flavonoids. The possible reaction mechanism for the reduction of Ag + has been proposed and discussed. The time-dependent electron micrographs and the simulation studies indicated that a physical interaction between the silver nanoparticles and the bacterial cell membrane may be responsible for this effect. Based on the findings, it seems very reasonable to believe that this greener way of synthesizing silver nanoparticles is not just an environmentally viable technique but it also opens up scope to improve their antibacterial properties.

  18. Spectroscopic and magnetic studies of highly dispersible superparamagnetic silica coated magnetite nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Tadyszak, Krzysztof [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Institute of Molecular Physics Polish Academy of Sciences, ul. Mariana Smo.luchowskiego 17, 60-179 Poznań (Poland); Kertmen, Ahmet, E-mail: ahmet.kertmen@pg.gda.pl [Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk (Poland); Coy, Emerson [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Andruszkiewicz, Ryszard; Milewski, Sławomir [Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk (Poland); Kardava, Irakli; Scheibe, Błażej; Jurga, Stefan [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Chybczyńska, Katarzyna, E-mail: katarzyna.chybczynska@ifmpan.poznan.pl [Institute of Molecular Physics Polish Academy of Sciences, ul. Mariana Smo.luchowskiego 17, 60-179 Poznań (Poland)

    2017-07-01

    Highlights: • Superparamagnetic core-shell nanoparticles of Fe{sub 2}O{sub 3}@Silica were obtained. • Magnetic response was studied by DC, AC magnetometry and EPR spectroscopy. • Nanoparticles show magnetite structure with a well-defined Verwey transition. • Samples show no inter particle magnetic interactions or agglomeration. - Abstract: Superparamagnetic behavior in aqueously well dispersible magnetite core-shell Fe{sub 3}O{sub 4}@SiO{sub 2} nanoparticles is presented. The magnetic properties of core-shell nanoparticles were measured with use of the DC, AC magnetometry and EPR spectroscopy. Particles where characterized by HR-TEM and Raman spectroscopy, showing a crystalline magnetic core of 11.5 ± 0.12 nm and an amorphous silica shell of 22 ± 1.5 nm in thickness. The DC, AC magnetic measurements confirmed the superparamagnetic nature of nanoparticles, additionally the EPR studies performed at much higher frequency than DC, AC magnetometry (9 GHz) have confirmed the paramagnetic nature of the nanoparticles. Our results show the excellent magnetic behavior of the particles with a clear magnetite structure, which are desirable properties for environmental remediation and biomedical applications.

  19. Study of mechanism of enhanced antibacterial activity by green synthesis of silver nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Parashar, Upendra Kumar; Srivastava, Sunil K; Srivastava, Anchal [Department of Physics, Banaras Hindu University, Varanasi 221005 (India); Kumar, Vinod; Saxena, Preeti S [Department of Zoology, Banaras Hindu University, Varanasi 22005 (India); Bera, Tanmay [Department of Mechanical, Materials and Aerospace Engineering, University of Central Florida, Orlando, FL 32816 (United States); Nath, Gopal [Department of Microbiology, Institute of Medical Science, Banaras Hindu University, Varanasi 22005 (India); Giri, Rajiv, E-mail: anchalbhu@gmail.com [Department of Materials Science and Engineering, Norwegian University of Science and Technology, NO-7491 Trondheim (Norway)

    2011-10-14

    The extensive use of silver nanoparticles needs a synthesis process that is greener without compromising their properties. The present study describes a novel green synthesis of silver nanoparticles using Guava (Psidium guajava) leaf extract. In order to compare with the conventionally synthesized ones, we also prepared Ag-NPs by chemical reduction. Their optical and morphological characteristics were thoroughly investigated and tested for their antibacterial properties on Escherichia coli. The green synthesized silver nanoparticles showed better antibacterial properties than their chemical counterparts even though there was not much difference between their morphologies. Fourier transform infrared (FTIR) spectroscopic analysis of the used extract and as-synthesized silver nanoparticles suggests the possible reduction of Ag{sup +} by the water-soluble ingredients of the guava leaf like tannins, eugenol and flavonoids. The possible reaction mechanism for the reduction of Ag{sup +} has been proposed and discussed. The time-dependent electron micrographs and the simulation studies indicated that a physical interaction between the silver nanoparticles and the bacterial cell membrane may be responsible for this effect. Based on the findings, it seems very reasonable to believe that this greener way of synthesizing silver nanoparticles is not just an environmentally viable technique but it also opens up scope to improve their antibacterial properties.

  20. A comparative study of TiO2 nanoparticles synthesized in premixed and diffusion flames

    Science.gov (United States)

    Ma, Hsiao-Kang; Yang, Hsiung-An

    2010-12-01

    Previous studies have been shown that synthesis of titania (TiO2) crystalline phase purity could be effectively controlled by the oxygen concentration through titanium tetra-isopropoxide (TTIP) via premixed flame from a Bunsen burner. In this study, a modified Hencken burner was used to synthesize smaller TiO2 nanoparticles via short diffusion flames. The frequency of collisions among particles would decrease and reduce TiO2 nanoparticle size in a short diffusion flame height. The crystalline structure of the synthesized nanoparticles was characterized by X-ray diffraction (XRD), transmission electron microscope (TEM), Barrett-Joyner-Halenda (BJH) and Brunauer-Emmett-Teller (BET) measurements. The characteristic properties of TiO2 nanoparticles synthesized from a modified Hencken burner were compared with the results from a Bunsen burner and commercial TiO2 (Degussa P25). The results showed that the average particle size of 6.63 nm from BET method was produced by a modified Hencken burner which was smaller than the TiO2 in a Bunsen burner and commercial TiO2. Moreover, the rutile content of TiO2 nanoparticles increased as the particle collecting height increased. Also, the size of TiO2 nanoparticles was highly dependent on the TTIP loading and the collecting height in the flame.

  1. Comprehensive studies on the interactions between chitosan nanoparticles and some live cells

    International Nuclear Information System (INIS)

    Zheng Aiping; Liu Huixue; Yuan Lan; Meng Meng; Wang Jiancheng; Zhang Xuan; Zhang Qiang

    2011-01-01

    As more and more oral formulations of nanoparticles are used in clinical contexts, a comprehensive study on the mechanisms of interaction between polymer nanoparticles and live cells seems merited. Such a study was conducted and the results were compared to the polymer itself in order to demonstrate different kinds of effects that are brought into the cell by polymer and its nanoparticles, especially the effects on the biomembrane. Several techniques, including surface plasmon resonance (SPR), Fourier transformed infrared spectroscopy (FTIR), Raman spectroscopy, fluorescence polarization spectroscopy (FP), flow cytometry (FCM) with quantitative analysis, and confocal images with antibody staining were employed toward this end. The cytotoxicity in vitro was also evaluated. Chitosan (CS), a polycationic polymer, was used to prepare the nanoparticles. We demonstrate that chitosan nanoparticles (CS-NP) induce strong alterations in the distribution of membrane proteins, fluidity of membrane lipids, and general membrane structure. Furthermore, the uptake of CS-NP into Caco-2 cells was found to have a similar mechanism to that of CS molecules, but the differences in degree were noted. These results indicate that positive charge and nanoscale size were the factors that most significantly affected the interactions between the nanoparticles of polycationic polymers and live cells. However, no difference in cytotoxicity toward the Caco-2 cells was found between CS and CS-NP. This supports the idea that CS-NP is an effective and safe carrier for oral drug delivery.

  2. A spectroscopic study on the interaction between gold nanoparticles and hemoglobin

    International Nuclear Information System (INIS)

    Garabagiu, Sorina

    2011-01-01

    Highlights: ► The interaction was studied using UV–vis and fluorescence spectroscopy. ► Gold nanoparticles quench the fluorescence emission of hemoglobin solution. ► The binding and thermodynamic constants were calculated. ► Major impact: electrochemical applications of the complex onto a substrate. -- Abstract: The interaction between horse hemoglobin and gold nanoparticles was studied using optical spectroscopy. UV–vis and fluorescence spectra show that a spontaneous binding process occurred between hemoglobin and gold nanoparticles. The Soret band of hemoglobin in the presence of gold nanoparticles does not show significant changes, which proves that the protein retained its biological function. A shift to longer wavelengths appears in the plasmonic band of gold nanoparticles upon the attachment of hemoglobin molecules. Gold nanoparticles quench the fluorescence emission of tryptophan residues in the structure of hemoglobin. The Stern–Volmer quenching constant, the binding constant and the number of binding sites were also calculated. Thermodynamic parameters indicate that the binding was mainly due to hydrophobic interactions.

  3. Nanobarcoding for improved nanoparticle detection in nanomedical biodistribution studies

    Science.gov (United States)

    Eustaquio, Trisha

    Determination of the fate of nanoparticles (NPs) in a biological system, or NP biodistribution, is critical in evaluating a NP formulation for nanomedicine. Unlike small-molecule drugs, NPs impose unique challenges in the design of appropriate biodistribution studies due to their small size and subsequent detection signal. Current methods to determine NP biodistribution are greatly inadequate due to their limited detection thresholds. There is an overwhelming need for a sensitive and efficient imaging-based method that can (1) detect and measure small numbers of NPs of various types, ideally single NPs, (2) associate preferential NP uptake with histological cell type by preserving spatial information in samples, and (3) allow for relatively quick and accurate NP detection in in vitro (and possibly ex vivo) samples for comprehensive NP biodistribution studies. Herein, a novel method for improved NP detection is proposed, coined "nanobarcoding." Nanobarcoding utilizes a non-endogenous oligonucleotide, or "nanobarcode" (NB), conjugated to the NP surface to amplify the detection signal from a single NP via in situ polymerase chain reaction (ISPCR), and this signal amplification will facilitate rapid and precise detection of single NPs inside cells over large areas of sample such that more sophisticated studies can be performed on the NP-positive subpopulation. Moreover, nanobarcoding has the potential to be applied to the detection of more than one NP type to study the effects of physicochemical properties, targeting mechanisms, and route of entry on NP biodistribution. The nanobarcoding method was validated in vitro using NB-functionalized superparamagnetic iron oxide NPs (NB-SPIONs) as the model NP type for improved NP detection inside HeLa human cervical cancer cells, a cell line commonly used for ISPCR-mediated detection of human papilloma virus (HPV). Nanotoxicity effects of NB-SPIONs were also evaluated at the single-cell level using LEAP (Laser-Enabled Analysis

  4. Albumin pre-coating enhances intracellular siRNA delivery of multifunctional amphiphile/siRNA nanoparticles

    Science.gov (United States)

    Kummitha, China M; Malamas, Anthony S; Lu, Zheng-Rong

    2012-01-01

    Nonspecific association of serum molecules with short-interfering RNA (siRNA) nanoparticles can change their physiochemical characteristics, and results in reduced cellular uptake in the target tissue during the systemic siRNA delivery process. Serum albumin is the most abundant protein in the body and has been used to modify the surface of nanoparticles, to inhibit association of other serum molecules. Here, we hypothesized that surface modification of lipid-based nanoparticular siRNA delivery systems with albumin could prevent their interaction with serum proteins, and improve intracellular uptake. In this study, we investigated the influence of albumin on the stability and intracellular siRNA delivery of the targeted siRNA nanoparticles of a polymerizable and pH-sensitive multifunctional surfactant N-(1-aminoethyl) iminobis[N-(oleoylcysteinylhistinyl-1-aminoethyl)propionamide] (EHCO) in serum. Serum resulted in a significant increase in the size of targeted EHCO/siRNA nanoparticles and inhibited cellular uptake of the nanoparticles. Coating of targeted EHCO/siRNA nanoparticles with bovine serum albumin at 9.4 μM prior to cell transfection improved cellular uptake and gene silencing efficacy of EHCO/siRNA targeted nanoparticles in serum-containing media, as compared with the uncoated nanoparticles. At a proper concentration, albumin has the potential to minimize interactions of serum proteins with siRNA nanoparticles for effective systemic in vivo siRNA delivery. PMID:23055731

  5. Delivery of human NKG2D-IL-15 fusion gene by chitosan nanoparticles to enhance antitumor immunity

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Chen; Jie, Leng; Yongqi, Wang [Department of Immunology, School of Medicine, Yangzhou University, Yangzhou, 225009 (China); Weiming, Xiao [Department of Gastroenterology, The Second Clinical Medical College, Yangzhou University, Yangzhou, 225009 (China); Juqun, Xi [Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou, 225009 (China); Yanbing, Ding [Department of Gastroenterology, The Second Clinical Medical College, Yangzhou University, Yangzhou, 225009 (China); Li, Qian [Department of Immunology, School of Medicine, Yangzhou University, Yangzhou, 225009 (China); Xingyuan, Pan [Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, 225009 (China); Mingchun, Ji [Department of Immunology, School of Medicine, Yangzhou University, Yangzhou, 225009 (China); Weijuan, Gong, E-mail: wjgong@yzu.edu.cn [Department of Immunology, School of Medicine, Yangzhou University, Yangzhou, 225009 (China); Department of Gastroenterology, The Second Clinical Medical College, Yangzhou University, Yangzhou, 225009 (China); Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou, 225009 (China); Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, 225009 (China); Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009 (China)

    2015-07-31

    Nanoparticles are becoming promising carriers for gene delivery because of their high capacity in gene loading and low cell cytotoxicity. In this study, a chitosan-based nanoparticle encapsulated within a recombinant pcDNA3.1-dsNKG2D-IL-15 plasmid was generated. The fused dsNKG2D-IL-15 gene fragment consisted of double extracellular domains of NKG2D with IL-15 gene at downstream. The average diameter of the gene nanoparticles ranged from 200 nm to 400 nm, with mean zeta potential value of 53.8 ± 6.56 mV. The nanoparticles which were loaded with the dsNKG2D-IL-15 gene were uptaken by tumor cells with low cytotoxicity. Tumor cells pre-transfected by gene nanopartilces stimulated NK and T cells in vitro. Intramuscular injection of gene nanoparticles suppressed tumor growth and prolonged survival of tumor-bearing mice through activation of NK and CD8{sup +} T cells. Thus, chitosan-based nanoparticle delivery of dsNKG2D-IL-15 gene vaccine can be potentially used for tumor therapy. - Highlights: • Generation of a nanoparticle for delivery of dsNKG2D-IL-15 gene. • Characterization of the gene nanoparticle. • Antitumor activity mediated by the gene nanoparticle.

  6. Synthesis and cytotoxicity study of magnesium ferrite-gold core-shell nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Nonkumwong, Jeeranan [Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Pakawanit, Phakkhananan [Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Wipatanawin, Angkana [Division of Biochemistry and Biochemical Technology, Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Jantaratana, Pongsakorn [Department of Physics, Faculty of Science, Kasetsart University, Bangkok 11900 (Thailand); Ananta, Supon [Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Srisombat, Laongnuan, E-mail: slaongnuan@yahoo.com [Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand)

    2016-04-01

    In this work, the core-magnesium ferrite (MgFe{sub 2}O{sub 4}) nanoparticles were prepared by hydrothermal technique. Completed gold (Au) shell coating on the surfaces of MgFe{sub 2}O{sub 4} nanoparticles was obtained by varying core/shell ratios via a reduction method. Phase identification, morphological evolution, optical properties, magnetic properties and cytotoxicity to mammalian cells of these MgFe{sub 2}O{sub 4} core coated with Au nanoparticles were examined by using a combination of X-ray diffraction, scanning electron microscopy, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy, UV–visible spectroscopy (UV–vis), vibrating sample magnetometry and resazurin microplate assay techniques. In general, TEM images revealed different sizes of the core-shell nanoparticles generated from various core/shell ratios and confirmed the completed Au shell coating on MgFe{sub 2}O{sub 4} core nanoparticles via suitable core/shell ratio with particle size less than 100 nm. The core-shell nanoparticle size and the quality of coating influence the optical properties of the products. The UV–vis spectra of complete coated MgFe{sub 2}O{sub 4}-Au core-shell nanoparticles exhibit the absorption bands in the near-Infrared (NIR) region indicating high potential for therapeutic applications. Based on the magnetic property measurement, it was found that the obtained MgFe{sub 2}O{sub 4}-Au core-shell nanoparticles still exhibit superparamagnetism with lower saturation magnetization value, compared with MgFe{sub 2}O{sub 4} core. Both of MgFe{sub 2}O{sub 4} and MgFe{sub 2}O{sub 4}-Au core-shell also showed in vitro non-cytotoxicity to mouse areola fibroblast (L-929) cell line. - Highlights: • Synthesis of MgFe{sub 2}O{sub 4}-Au core-shell nanoparticles with particle size < 100 nm • Complete Au shell coating on the surfaces of MgFe{sub 2}O{sub 4} nanoparticles • In vitro cytotoxicity study of complete coated MgFe{sub 2}O{sub 4}-Au core

  7. Marked transfection enhancement by the DPL (DNA/peptide/lipid) complex.

    Science.gov (United States)

    Moon, Ik-Jae; Kang, Hyungu; Seu, Young-Bae; Chang, Byeong-Churl; Song, Dae-Kyu; Park, Jong-Gu

    2007-10-01

    A short peptide, corresponding to the nuclear localization signal of the human immunodeficiency virus-1 Tat protein, Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg, was modified by adding a cysteine residue at the COOH terminus. The peptide was mixed with a reporter plasmid, and then with cationic lipids, to form a tripartite complex, DNA/peptide/lipid (DPL). Various cell lines were treated with the DPL complex and compared for transfection efficiency with those of the conventional DNA/lipid (DL) complex. With the simple inclusion of the peptide, the DPL complex showed much enhanced transfection. Meanwhile, the plasmid DNA mixed only with the peptide exhibited some improvement but with much lower transfection than the DPL complex. When the DPL complex was formed with various cationic lipids, the DOSPA/DOPE exhibited superior transfection efficiency than the other cationic lipids tested at the optimal ratio of 1:3:5 (w:w:w) in many cell types. At the optimal ratio of the DPL components, transfection efficiency was routinely shown to be approximately 10-fold higher for reporter gene expression than that of the conventional DL complex. Furthermore, when subcutaneous tumors of a colon cancer cell line (SW480) were treated intratumorally with antisense oligos, k-ras-RiAS, delivered as a DPL complex, tumor growth was markedly suppressed. This study shows that the DPL complex, which is easy to formulate by ordered mixing, can be employed for a much enhanced cellular uptake of a transgene both in vitro and in vivo.

  8. Gene therapy of transplant arteriopathy by liposome-mediated transfection of endothelial nitric oxide synthase.

    Science.gov (United States)

    Iwata, A; Sai, S; Moore, M; Nyhuis, J; de Fries-Hallstrand, R; Quetingco, G C; Allen, M D

    2000-11-01

    Transplant arteriopathy is the major factor limiting long-term survival after cardiac transplantation. We have previously demonstrated that liposome-mediated gene delivery of endothelial nitric oxide synthase (eNOS) to donor hearts reduces ischemia-reperfusion injury by blocking NFkappaB activation, adhesion molecule expression, and leukocyte infiltration. In this study, we used gene transfer of eNOS in a rabbit carotid transplant model to see whether these same effects would similarly ameliorate transplant arteriopathy. Liposomes complexed to the gene encoding eNOS were injected into donor carotid arterial segments that were transplanted orthotopically into recipient carotid arteries (n = 10). Controls included transplanted carotids transfected with liposomes complexed to empty plasmids (no functional gene) (n = 4) and transplanted carotids treated with saline (n = 6). Transplanted arteries were harvested for processing at 21 days. Intima/media (I/M) area ratios were calculated by computerized image analysis. Infiltrating T-lymphocytes and macrophages, and expression of VCAM-1 and ICAM-1 were quantified on immunocytochemistry. The I/M ratio was significantly reduced in eNOS-transfected arteries compared with arteries transfected with empty plasmids and saline-treated controls. Compared to transplanted control arteries, eNOS-transfected arteries demonstrated significantly reduced T-cell infiltration into the intima and significantly reduced macrophage infiltration into the media. Cell surface expression of VCAM-1 and ICAM-1 were both reduced in eNOS-transfected arteries. ENOS gene delivery can suppress neointimal lesion formation and T-lymphocyte and macrophage infiltration in transplanted arteries, associated with a reduction in relevant adhesion molecule expression. Thus, gene therapy with eNOS may not only reduce ischemia-reperfusion injury but may also ameliorate transplant arteriopathy in transplanted hearts.

  9. Transfection of primary brain capillary endothelial cells for protein synthesis and secretion of recombinant erythropoietin: a strategy to enable protein delivery to the brain

    DEFF Research Database (Denmark)

    Burkhart, Annette; Andresen, Thomas Lars; Aigner, Achim

    2017-01-01

    , as turning BCECs into recombinant protein factories by transfection could result in protein secretion further into the brain. The present study aims to investigate the possibility of transfecting primary rat brain endothelial cells (RBECs) for recombinant protein synthesis and secretion...... of the neuroprotective protein erythropoietin (EPO). We previously showed that 4% of RBECs with BBB properties can be transfected without disrupting the BBB integrity in vitro, but it can be questioned whether this is sufficient to enable protein secretion at therapeutic levels. The present study examined various...... transfection vectors, with regard to increasing the transfection efficiency without disrupting the BBB integrity. Lipofectamine 3000™ was the most potent vector compared to polyethylenimine (PEI) and Turbofect. When co-cultured with astrocytes, the genetically modified RBECs secreted recombinant EPO...

  10. Silver Nanoparticles

    Science.gov (United States)

    Khaydarov, R. R.; Khaydarov, R. A.; Estrin, Y.; Evgrafova, S.; Scheper, T.; Endres, C.; Cho, S. Y.

    The bactericidal effect of silver nanoparticles obtained by a novel electrochemical method on Escherichia coli, Staphylococcus aureus, Aspergillus niger and Penicillium phoeniceum cultures has been studied. The tests conducted have demonstrated that synthesized silver nanoparticles — when added to water paints or cotton fabrics — show a pronounced antibacterial/antifungal effect. It was shown that smaller silver nanoparticles have a greater antibacterial/antifungal efficacy. The paper also provides a review of scientific literature with regard to recent developments in the field of toxicity of silver nanoparticles and its effect on environment and human health.

  11. Simulation study of depositing the carbon film on nanoparticles in the magnetized methane plasma

    Science.gov (United States)

    Mohammadzadeh, Hosein; Pourali, Nima; Ebadi, Zahra

    2018-03-01

    Plasma coating of nanoparticles in low-temperature magnetized methane plasma is studied by a simulation approach. To this end, by using the global model, the electron temperature and concentration of different species considered in this plasma are determined in the center of a capacitively coupled discharge. Then, the plasma-wall transition region in the presence of an oblique magnetic field is simulated by the multi-component fluid description. Nanoparticles with different radii are injected into the transition region and surface deposition and heating models, as well as dynamics and charging models, are employed to examine the coating process. The results of the simulation show that the non-spherical growth of nanoparticles is affected by the presence of the magnetic field, as with passing time, an oscillating increase is seen in the thickness of the film deposited on nanoparticles. Also, it is shown that the uniformity of the deposited film is dependent on the rotation velocity of nanoparticles. Generally, the obtained results imply that the sphericity of nanoparticles and uniformity of the film coated on them are controllable by the magnitude and orientation of the magnetic field.

  12. Mechanistic Study of Silver Nanoparticle's Synthesis by Dragon's Blood Resin Ethanol Extract and Antiradiation Activity.

    Science.gov (United States)

    Hasan, Murtaza; Iqbal, Javed; Awan, Umer; Saeed, Yasmeen; Ranran, Yuan; Liang, Yanli; Dai, Rongji; Deng, Yulin

    2015-02-01

    Biological synthesis of nanoparticles is best way to avoid exposure of hazardous materials as compared to chemical manufacturing process which is a severe threat not only to biodiversity but also to environment. In present study, we reported a novel method of finding antiradiation compounds by bioreducing mechanism of silver nanoparticles formation using 50% ethanol extract of Dragons blood, a famous Chinese herbal plant. Color change during silver nanoparticles synthesis was observed and it was confirmed by ultra violet (UV) visible spectroscopy at wave length at 430 nm after 30 min of reaction at 60 °C. Well dispersed round shaped silver nanoparticles with approximate size (4 nm to 50 nm) were measured by TEM and particle size analyser. Capping of biomolecules on Ag nanoparticles was characterized by FTIR spectra. HPLC analysis was carried out to find active compounds in the extract. Furthermore, antiradiation activity of this extract was tested by MTT assay in vitro after incubating the SH-SY5Y cells for 24 h at 37 °C. The results indicate that presence of active compounds in plant extract not only involves in bioreduction process but also shows response against radiation. The dual role of plant extract as green synthesis of nanoparticles and exhibit activity against radiation which gives a new way of fishing out active compounds from complex herbal plants.

  13. Studying the Adsorption Process of Riboflavin on Silver-Deposited Fe3O4 Nanoparticles

    Directory of Open Access Journals (Sweden)

    Morteza Akhond

    2016-12-01

    Full Text Available The adsorption characteristics of riboflavin onto silver-deposited iron oxide magnetic nanoparticles (Ag/Fe3O4 have been described. Characterization of the synthesized Ag/Fe3O4 nanoparticles was achieved by FTIR spectra, TEM image and XRD pattern. The influence of several experimental parameters such as nanoparticles dosage, pH of the sample solution, different orientations of the riboflavin molecules toward Ag/Fe3O4 surface, riboflavin concentration, contact time of the reagents, temperature, ionic strength and presence of halide anions were studied. Experimental data indicated that Ag/Fe3O4 nanoparticles adsorb more than 90% of riboflavin under the optimum experimental conditions of the adsorbent dosage of 4.0 mg, a pH of 6.0, and a contact time of 2.0 min, when an initial riboflavin concentration of 0.02 mM is used. The results revealed that the presence of halide anions lower the adsorption of riboflavin on the surface of nanoparticles due to dissolution of the silver layer of the nanoparticles. It was found that the adsorption isotherm is best fitted to Dubinin-Radushkevich and Freundlich models and kinetic model followed a pseudo-second-order adsorption rate.

  14. Comparative study on the uptake and bioimpact of metal nanoparticles released into environment

    Science.gov (United States)

    Andries, Maria; Pricop, Daniela; Grigoras, Marian; Lupu, Nicoleta; Sacarescu, Liviu; Creanga, Dorina; Iacomi, Felicia

    2015-12-01

    Metallic particles of very small size are ubiquitously released in the air, water and soil from various natural and artificial sources - the last ones with enhanced extent since nanotechnology development accelerated exponentially. In this study we focused on the impact of metal nanoparticles in vegetal species of agroindustrial interest namely the maize (Zea mais L.). Laboratory simulation of environmental pollution was carried out by using engineered nanoparticles of two types: iron oxides with magnetic properties and gold nanoparticles supplied in the form of dilutes stable suspensions in the culture medium of maize seedlings. Magnetic nanoparticle (MNPs) preparation was performed by applying chemical route from iron ferric and ferrous precursor salts in alkali reaction medium at relatively high temperature (over 80 °C). Gold nanoparticles (GNPs) synthesis was accomplished from auric hydrochloride acid in alkali reaction medium in similar temperature conditions. In both types of metallic nanoparticles citrate ions were used as coating shell with role of suspension stabilization. Plantlet response was assessed at the level of assimilatory pigment contents in green tissue of seedlings in early ontogenetic stages.

  15. HRTEM Study of the Role of Nanoparticles in ODS Ferritic Steel

    Energy Technology Data Exchange (ETDEWEB)

    Hsiung, L; Tumey, S; Fluss, M; Serruys, Y; Willaime, F

    2011-08-30

    Structures of nanoparticles and their role in dual-ion irradiated Fe-16Cr-4.5Al-0.3Ti-2W-0.37Y{sub 2}O{sub 3} (K3) ODS ferritic steel produced by mechanical alloying (MA) were studied using high-resolution transmission electron microscopy (HRTEM) techniques. The observation of Y{sub 4}Al{sub 2}O{sub 9} complex-oxide nanoparticles in the ODS steel imply that decomposition of Y{sub 2}O{sub 3} in association with internal oxidation of Al occurred during mechanical alloying. HRTEM observations of crystalline and partially crystalline nanoparticles larger than {approx}2 nm and amorphous cluster-domains smaller than {approx}2 nm provide an insight into the formation mechanism of nanoparticles/clusters in MA/ODS steels, which we believe involves solid-state amorphization and re-crystallization. The role of nanoparticles/clusters in suppressing radiation-induced swelling is revealed through TEM examinations of cavity distributions in (Fe + He) dual-ion irradiated K3-ODS steel. HRTEM observations of helium-filled cavities (helium bubbles) preferably trapped at nanoparticle/clusters in dual-ion irradiated K3-ODS are presented.

  16. Study of hematite-iron phase transformation during iron-carbon core-shell nanoparticles synthesis and investigation of their magnetic and microwave properties

    OpenAIRE

    Omid Khani; Morteza Zargar Shoushtari; Mohammad Jazirehpour; Mansoor Farbod

    2017-01-01

    The structural properties and microwave absorption capability of the iron nanoparticles and iron-carbon core-shell nanoparticles have been studied, in the present paper. The investigated nanoparticles were synthesized by hydrothermal route and by reduction of hematite nanoparticles during annealing in argon-hydrogen atmosphere. Hematite-iron phase transformation during the reduction process has been studied by X-ray diffraction (XRD). XRD patterns showed that in iron nanoparticles, hematite-i...

  17. Raman scattering studies on PEG functionalized hydroxyapatite nanoparticles.

    Science.gov (United States)

    Yamini, D; Devanand Venkatasubbu, G; Kumar, J; Ramakrishnan, V

    2014-01-03

    The pure hydroxyapatite (HAP) nanoparticles (NPs) have been synthesized by wet chemical precipitation method. Raman spectral measurements have been made for pure HAP, pure Polyethylene glycol (PEG) 6000 and PEG coated HAP in different mass ratios (sample 1, sample 2 and sample 3). The peaks observed in Raman spectrum of pure HAP and the XRD pattern have confirmed the formation of HAP NPs. Vibrational modes have been assigned for pure HAP and pure PEG 6000. The observed variation in peak position of Raman active vibrational modes of PEG in PEG coated HAP has been elucidated in this work, in terms of intermolecular interactions between PEG and HAP. Further these results suggest that the functionalization of nanoparticles may be independent of PEG mass. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. SANS study of multilayer nanoparticles based on block copolymer micelles

    Czech Academy of Sciences Publication Activity Database

    Pleštil, Josef; Pospíšil, Herman; Kadlec, Petr; Tuzar, Zdeněk; Kříž, Jaroslav; Gordeliy, V. I.

    2001-01-01

    Roč. 42, č. 7 (2001), s. 2941-2946 ISSN 0032-3861 R&D Projects: GA AV ČR KSK2050602; GA ČR GA203/97/0249; GA ČR GA203/00/1317 Institutional research plan: CEZ:AV0Z4050913 Keywords : small-angle neutron scattering * multilayer nanoparticles * coated micellar cores Subject RIV: CD - Macromolecular Chemistry Impact factor: 1.681, year: 2001

  19. Density functional theory studies of transition metal nanoparticles in catalysis

    DEFF Research Database (Denmark)

    Greeley, Jeffrey Philip; Rankin, Rees; Zeng, Zhenhua

    2013-01-01

    Periodic Density Functional Theory calculations are capable of providing powerful insights into the structural, energetics, and electronic phenomena that underlie heterogeneous catalysis on transition metal nanoparticles. Such calculations are now routinely applied to single crystal metal surfaces...... and electrocatalysis, in which single crystal models are combined with Wulff construction-based ideas to produce descriptions of average nanocatalyst behavior. Then, I will proceed to describe explicitly DFT-based descriptions of catalysis on truly nanosized particles (

  20. Computational studies of steering nanoparticles with magnetic gradients

    Science.gov (United States)

    Aylak, Sultan Suleyman

    Magnetic Resonance Imaging (MRI) guided nanorobotic systems that could perform diagnostic, curative, and reconstructive treatments in the human body at the cellular and subcellular level in a controllable manner have recently been proposed. The concept of a MRI-guided nanorobotic system is based on the use of a MRI scanner to induce the required external driving forces to guide magnetic nanocapsules to a specific target. However, the maximum magnetic gradient specifications of existing clinical MRI systems are not capable of driving magnetic nanocapsules against the blood flow. This thesis presents the visualization of nanoparticles inside blood vessel, Graphical User Interface (GUI) for updating file including initial parameters and demonstrating the simulation of particles and C++ code for computing magnetic forces and fluidic forces. The visualization and GUI were designed using Virtual Reality Modeling Language (VRML), MATLAB and C#. The addition of software for MRI-guided nanorobotic system provides simulation results. Preliminary simulation results demonstrate that external magnetic field causes aggregation of nanoparticles while they flow in the vessel. This is a promising result --in accordance with similar experimental results- and encourages further investigation on the nanoparticle-based self-assembly structures for use in nanorobotic drug delivery.

  1. Modulation of microfilament protein composition by transfected cytoskeletal actin genes

    Energy Technology Data Exchange (ETDEWEB)

    Ng, S.Y.; Erba, H.; Latter, G.; Kedes, L.; Leavitt, J.

    1988-04-01

    HuT-14T is a highly tumorigenic fibroblast cell line which exhibits a reduced steady-state level of ..beta..-actin due to coding mutations in one of two ..beta..-actin alleles. The normal rate of total actin synthesis could be restored in some clones of cells following transfection of the functional ..beta..-actin gene but not following transfection of the functional ..gamma..-actin gene. In ..gamma..-actin gene-transfected substrains that have increased rates of ..gamma..-actin synthesis, ..beta..-actin synthesis is further reduced in a manner consistent with an autoregulatory mechanism, resulting in abnormal ratios of actin isoforms. Thus, both ..beta..- and ..gamma..-actin proteins can apparently regulate the synthesis of their coexpressed isoforms. In addition, decreased synthesis of normal ..beta..-actin seems to correlate with a concomitant down-regulation of tropomyosin isoforms.

  2. Epizone: Interlaboratory Ring Trial to Compare Dna Transfection Efficiencies

    DEFF Research Database (Denmark)

    Dory, Daniel; Albina, Emmanuel; Kwiatek, Olivier

    of viruses by reverse genetics and/or generation of mutated viruses. A large number of transfection chemicals like calcium phospate, branched organic compounds, liposomes, cationic polymers etc. are available on the market which are used by different laboratories for different cell lines. To obtain...... an overview on the efficiencies of varying transfection procedures, an interlaboratory ring trial was initiated within EPIZONE theme 5. A total of 15 participitating laboratories from 7 member institutions received RK13 cells, plasmid DNA encoding firefly luciferase under the transcriptional control...... of the human cytomegalovirus major immediate early promoter, a specially developed lysis buffer and a detailed protocol. Transfected cells were harvested in the laboratories of the participants, frozen and sent to the FLI where both the luciferase activity and protein content of the individual samples were...

  3. Nucleic acid transfection and transgenesis in parasitic nematodes.

    Science.gov (United States)

    Lok, James B

    2012-04-01

    Transgenesis is an essential tool for assessing gene function in any organism, and it is especially crucial for parasitic nematodes given the dwindling armamentarium of effective anthelmintics and the consequent need to validate essential molecular targets for new drugs and vaccines. Two of the major routes of gene delivery evaluated to date in parasitic nematodes, bombardment with DNA-coated microparticles and intragonadal microinjection of DNA constructs, draw upon experience with the free-living nematode Caenorhabditis elegans. Bombardment has been used to transiently transfect Ascaris suum, Brugia malayi and Litomosoides sigmodontis with both RNA and DNA. Microinjection has been used to achieve heritable transgenesis in Strongyloides stercoralis, S. ratti and Parastrongyloides trichosuri and for additional transient expression studies in B. malayi. A third route of gene delivery revisits a classic method involving DNA transfer facilitated by calcium-mediated permeabilization of recipient cells in developing B. malayi larvae and results in transgene inheritance through host and vector passage. Assembly of microinjected transgenes into multi-copy episomal arrays likely results in their transcriptional silencing in some parasitic nematodes. Methods such as transposon-mediated transgenesis that favour low-copy number chromosomal integration may remedy this impediment to establishing stable transgenic lines. In the future, stable transgenesis in parasitic nematodes could enable loss-of-function approaches by insertional mutagenesis, in situ expression of inhibitory double-stranded RNA or boosting RNAi susceptibility through heterologous expression of dsRNA processing and transport proteins.

  4. Cell type and transfection reagent-dependent effects on viability, cell content, cell cycle and inflammation of RNAi in human primary mesenchymal cells

    DEFF Research Database (Denmark)

    Yang, Hsiao Yin; Vonk, Lucienne A.; Licht, Ruud

    2014-01-01

    The application of RNA interference (RNAi) has great therapeutic potential for degenerative diseases of cartilaginous tissues by means of fine tuning the phenotype of cells used for regeneration. However, possible non-specific effects of transfection per se might be relevant for future clinical...... application. In the current study, we selected two synthetic transfection reagents, a cationic lipid-based commercial reagent Lipofectamine RNAiMAX and polyethylenimine (PEI), and two naturally-derived transfection reagents, namely the polysaccharides chitosan (98% deacetylation) and hyaluronic acid (20...... 3 and day 6 post-transfection. In addition to silencing efficiency, non-specific effects such as cytotoxicity, change in DNA content and differentiation potential of cells were evaluated. Among the four transfection reagents, the commercial liposome-based agent was the most efficient reagent for si...

  5. Drug-polymer interaction studies of cytarabine loaded chitosan nanoparticles

    International Nuclear Information System (INIS)

    Madni, A.; Kashif, P.M.; Nazir, I.; Rehman, M.

    2017-01-01

    Assessment of possible incompatibilities between drug and excipients is an important parameter of preformulation stage during the pharmaceutical product development of active pharmaceutical ingredient (API). The potential physical and chemical interaction among the components of a delivery system can affect the chemical nature, bioavailability, stability, and subsequently therapeutic efficacy of drugs. In this study, ATR-FTIR spectroscopy was employed to investigate the possible intermolecular interaction of Cytarabine with deacetylated chitosan and tripolyphosphate in the resulting physical blends and crosslinked nanoparticulate system. Two different strategies, physical blending and ionotropic gelation, were adopted to prepare binary or tertiary mixtures and nanoparticulate formulation, respectively. The IR spectra of CB showed characteristic peaks at 3438.27 cm-1 (primary amine), 3264.74 cm-1 (hydroxyl group) and 1654.98 cm-1 (C=O stretch in cyclic ring); CS at 3361.47 cm-1 (N-H stretching), 1646.18 cm-1 (C=O of Amide I), 1582.36 cm-1 (C=O of Amide II), and sTPP at 1135.77 cm-1 (P=O). CS-sTPP chemical interaction was confirmed from the shift in the absorption band of carbonyl groups (amide I, II) to 1634.66 cm-1 and 1541.17 cm-1 in blank chitosan nanoparticles, and 1636.87 cm-1, 1543.33 cm-1 in CSNP1 (2:6:1), and at 1646.15 cm-1 and 1557.04 cm-1 in CSNP2 (1:3:1). The characteristic peaks of CB were also present in chitosan formulation with a slight shift in the amino group at 3429.43 cm-1 and 3423.21 cm-1, in the hydroxyl group at 3274.54 cm-1 and 3270.73 cm-1, CSNP1 and CSNP2, respectively. The findings counseled no significant interaction in IR absorption pattern of cytarabine functional groups after encapsulation in CS-sTPP complex, which projected the potential of chitosan nanoparticulate system to entrap cytarabine. (author)

  6. A simple, highly efficient method for heterologous expression in mammalian primary neurons using cationic lipid-mediated mRNA transfection

    Directory of Open Access Journals (Sweden)

    Damian J Williams

    2010-11-01

    Full Text Available Expression of heterologous proteins in adult mammalian neurons is a valuable technique for the study of neuronal function. The postmitotic nature of mature neurons prevents effective DNA transfection using simple, cationic lipid-based methods. Adequate heterologous protein expression is often only achievable using complex techniques that, in many cases, are associated with substantial toxicity. Here, a simple method for high efficiency transfection of mammalian primary neurons using in vitro-transcribed mRNA and the cationic lipid transfection reagent Lipofectamine 2000 is described. Optimal transfection conditions were established in adult mouse dissociated dorsal root ganglion (DRG neurons using a 96-well based luciferase activity assay. Using these conditions, a transfection efficiency of 25% was achieved in DRG neurons transfected with EGFP mRNA. High transfection efficiencies were also obtained in dissociated rat superior cervical ganglion (SCG neurons and mouse cortical and hippocampal cultures. Endogenous Ca2+ currents in EGFP mRNA-transfected SCG neurons were not significantly different from untransfected neurons, which suggested that this technique is well suited for heterologous expression in patch clamp recording experiments. Functional expression of a cannabinoid receptor (CB1R, a G protein inwardly-rectifying K+ channel (GIRK4 and a dominant-negative G protein α-subunit mutant (GoA G203T indicate that the levels of heterologous protein expression attainable using mRNA transfection are suitable for most functional protein studies. This study demonstrates that mRNA transfection is a straightforward and effective method for heterologous expression in neurons and is likely to have many applications in neuroscience research.

  7. Photochemically synthesized heparin-based silver nanoparticles: an antimicrobial activity study

    Science.gov (United States)

    Rodriguez-Torres, Maria del Pilar; Acosta-Torres, Laura Susana; Díaz-Torres, Luis Armando

    2017-08-01

    The antimicrobial activity of silver nanoparticles has been extensively studied in the last years. Such nanoparticles constitute a potential and promising approach for the development of new antimicrobial systems especially due to the fact that several microorganisms are developing resistance to some already existing antimicrobial agents, therefore making antibacterial and antimicrobial studies on alternative materials necessary to overcome this issue. Silver nanoparticle concentration and size are determining factors on the antimicrobial activity of these nano systems. Heparin is a polysaccharide that belongs to the glycosaminoglycans (GAGs) family, molecules formed by a base disaccharide whose components are joined by a glycosidic linkage that is a repeating unit along their structure. It is highly sulfated making it a negatively charged material that is also widely used as an anticoagulant in Medicine because its biocompatibility besides it is also produced within the human body, specifically in the mast cells. Heparin alone possesses antimicrobial activity although it has not been studied very much in detail, it only has been demonstrated that it inhibits E. coli, P. aeruginosa, S. aureus and S. epidermidis, so taking this into account, this study is dedicated to assess UV photochemically-synthesized (λ=254 nm) heparin-based silver nanoparticles antimicrobial activity using the agar disk diffusion method complemented by the broth microdilution method to estimate de minimum inhibitory concentration (MIC), that is the lowest concentration at which an antimicrobial will inhibit visible growth of a microorganism. The strains used were the ones aforementioned to assess the antimicrobial activity degree these heparinbased nanoparticles exhibit.

  8. Design of pH-sensitive peptides from natural antimicrobial peptides for enhancing polyethylenimine-mediated gene transfection.

    Science.gov (United States)

    Zhang, Shi-Kun; Song, Jin-Wen; Li, Su-Bo; Gao, Hong-Wei; Chang, Hong-Yu; Jia, Li-Li; Gong, Feng; Tan, Ying-Xia; Ji, Shou-Ping

    2017-05-01

    Poor endosomal release is a major barrier of polyplex-mediated gene transfection. Antimicrobial peptides (AMPs) are commonly used to improve polyethylenimine (PEI)-mediated gene transfection by increasing endosomal release. In the present study, we designed novel pH-sensitive peptides that highly enhance transfection efficiency compared to their parent peptides. Two analogues of melittin (Mel) and RV-23 (RV) were synthesized by replacing the positively-charged residues in their sequences with glutamic acid residues. The pH-sensitive lysis ability of the peptides, the effect of the peptides on physicochemical characteristics, the intracellular trafficking, the transfection efficiency, and the cytotoxicity of the polyplexes were determined. The acidic peptides showed pH-sensitive lytic activity. The hemolytic activity of acidic peptides at pH 5.0 was higher than that at pH 7.4. The incorporation of acidic peptides did not affect the DNA binding ability of PEI but affected the physicochemical characteristics of the PEI/DNA polyplexes, which may be beneficial for endosomal release and gene transfection. The incorporation of acidic peptides into PEI/DNA polyplexes enhanced the PEI-mediated transfection efficiency corresponding to up to 42-fold higher luciferase activity compared to that of PEI alone. The results of the present study indicate that replacement of positively-charged residues with glutamic acid residues in the AMP sequence yields pH-sensitive peptides, which enhance the transfection efficiency of PEI/DNA polyplexes in various cell lines. Copyright © 2017 John Wiley & Sons, Ltd.

  9. Role of Glycol Chitosan-incorporated Ursolic Acid Nanoparticles in ...

    African Journals Online (AJOL)

    Purpose: To investigate the effect of ursolic acid (UA)-incorporated glycol chitosan (GC) nanoparticles on inhibition of human osteosarcoma. Methods: U2OS and Saos-2 osteosarcoma cells were transfected with ursolic acid (UA) incorporated glycol chitosan (GC) nanoparticles. Ultraviolet (UV) spectrophotometry was used ...

  10. X-ray Spectromicroscopy Studies of Nanoparticles in the Environment

    Science.gov (United States)

    Sedlmair, J.; Gleber, S.-C.; Schirz, A.; Zanker, H.; Thieme, J.

    2009-04-01

    Motivation: In recent time, carbon nanotubes (CNTs) have drawn a lot of attention due to their unique properties and due to that possible application, for instance in pharmacology, material sciences or as semiconductors. CNTs are tubes with diameters in the nanometer scale, but with lengths up to several millimeters. Their walls consist of carbon atoms, each bound to three other carbon atoms (sp2-hybridization), which results in a hexagonal honeycomb-like structure. They can also be functionalized, e.g. with carboxyl- or hydroxyl groups. Although the production and modification of CNTs in sizable quantities have been improved continuously, the characterization of these nano-particles still needs to be advanced. Additionally, the ecological aspect comes into account. Since most of these new materials consist of particles too small to be biodegraded, it is important to analyze the impact of CNTs on the environment (and biomolecular matter in general). Here we present the result of a study of pristine and functionalized carbon nanotubes (CNTs) using the x-ray scanning transmission microscope (STXM) at the storage ring BESSY II in Berlin for a NEXAFS (near edge x-ray absorption spectroscopy) analysis with spatial resolution. Experiment and results: We characterized three types of multi-walled CNTs (3-15 walls, outer diameter of 13-16 nm and length distribution 1-10 nm) by x-ray spectromicroscopy. To be more specific, we have investigated different CNT-samples with energies around the C1s K-shell edge (~284 eV) dry and in aqueous environment at ambient conditions. Using the STXM, the spatial information from the x-ray image with a pixel size of 50 nm can be combined with NEXAFS-spectra[5] of each pixel of the image area. The differences between the species are observable both in the microscopic images and the spectral data. The evaluation[1][2] of the NEXAFS-spectra yields information about the chemical bindings in the sample. Discussion The difference between the

  11. Loading of atorvastatin and linezolid in β-cyclodextrin–conjugated cadmium selenide/silica nanoparticles: A spectroscopic study

    Energy Technology Data Exchange (ETDEWEB)

    Antony, Eva Janet; Shibu, Abhishek [Department of Nanosciences & Technology, Karunya University, Coimbatore 641114, Tamil Nadu (India); Ramasamy, Sivaraj; Paulraj, Mosae Selvakumar [Department of Chemistry, Karunya University, Coimbatore 641114, Tamil Nadu (India); Enoch, Israel V.M.V., E-mail: drisraelenoch@gmail.com [Department of Nanosciences & Technology, Karunya University, Coimbatore 641114, Tamil Nadu (India); Department of Chemistry, Karunya University, Coimbatore 641114, Tamil Nadu (India)

    2016-08-01

    The preparation of β–cyclodextrin–conjugated cadmium selenide–silica nanoparticles, the loading of two drugs viz., Atorvastatin and linezolid in the cyclodextrin cavity, and the fluorescence energy transfer between CdSe/SiO{sub 2} nanoparticles and the drugs encapsulated in the cyclodextrin cavity are reported in this paper. IR spectroscopy, X-ray diffractometry, transmission electron microscopy, and particle size analysis by light–scattering experiment were used as the tools of characterizing the size and the crystal system of the nanoparticles. The nanoparticles fall under hexagonal system. The silica–shell containing CdSe nanoparticles were functionalized by reaction with aminoethylamino–β–cyclodextrin. Fluorescence spectra of the nanoparticles in their free and drug–encapsulated forms were studied. The FÖrster distances between the encapsulated drugs and the CdSe nanoparticles are below 3 nm. The change in the FÖrster resonance energy parameters under physiological conditions may aid in tracking the release of drugs from the cavity of the cyclodextrin. - Highlights: • CdSe/SiO{sub 2} nanoparticles of crystallite size 15 nm are prepared. • β-Cyclodextrin is attached to the surface of the nanoparticles. • Atorvastatin and linezolid get encapsulated in the cyclodextrin cavity. • FRET efficiency between the nanoparticles and the loaded drugs are determined.

  12. Study of internalization and viability of multimodal nanoparticles for labeling of human umbilical cord mesenchymal stem cells

    International Nuclear Information System (INIS)

    Miyaki, Liza Aya Mabuchi; Sibov, Tatiana Tais; Pavon, Lorena Favaro; Mamani, Javier Bustamante; Gamarra, Lionel Fernel

    2012-01-01

    Objective: To analyze multimodal magnetic nanoparticles-Rhodamine B in culture media for cell labeling, and to establish a study of multimodal magnetic nanoparticles-Rhodamine B detection at labeled cells evaluating they viability at concentrations of 10 μg Fe/mL and 100μg Fe/mL. Methods: We performed the analysis of stability of multimodal magnetic nanoparticles-Rhodamine B in different culture media; the mesenchymal stem cells labeling with multimodal magnetic nanoparticles-Rhodamine B; the intracellular detection of multimodal magnetic nanoparticles-Rhodamine B in mesenchymal stem cells, and assessment of the viability of labeled cells by kinetic proliferation. Results: The stability analysis showed that multimodal magnetic nanoparticles-Rhodamine B had good stability in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium. The mesenchymal stem cell with multimodal magnetic nanoparticles-Rhodamine B described location of intracellular nanoparticles, which were shown as blue granules co-localized in fluorescent clusters, thus characterizing magnetic and fluorescent properties of multimodal magnetic nanoparticles Rhodamine B. Conclusion: The stability of multimodal magnetic nanoparticles-Rhodamine B found in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium assured intracellular mesenchymal stem cells labeling. This cell labeling did not affect viability of labeled mesenchymal stem cells since they continued to proliferate for five days. (author)

  13. Studies on the Genotoxicity Behavior of Silver Nanoparticles in the Presence of Heavy Metal Cadmium Chloride in Mice

    Directory of Open Access Journals (Sweden)

    Hanan Ramadan Hamad Mohamed

    2016-01-01

    Full Text Available Incredible rapid growth in the nanoparticles applications and development increases the daily human exposure to them but humans are exposed to many other pollutants in addition to nanoparticles that forced us to evaluate the effect of heavy metal cadmium chloride (CdCl2 coinjection on silver nanoparticles induced genotoxic risk in this study. Mice were injected into the abdominal cavity with single dose of Ag nanoparticles (20, 41, and 82 mg/kg or CdCl2 (1.5 mg/kg either separately or together simultaneously and sacrificed 24 hours later. CdCl2 cotreatment enhanced the induced dose-dependent sperm abnormality by Ag nanoparticles different doses as shown by the statistical significant decreases in both sperm concentration and motility and increases in the frequency of abnormal sperms and also potentiated the Ag nanoparticles induced chromosomal and DNA damage indicated by the statistical significant elevations in the frequencies of micronucleated polychromatic erythrocytes (MNPCEs and DNA damage levels. Moreover, statistical elevations in malondialdehyde level and reductions in catalase activity were observed after CdCl2 coinjection with Ag nanoparticles compared with Ag nanoparticles treated groups’ values. Ag nanoparticles induced sperm abnormality, clastogenicity, and genotoxicity were potentiated by heavy metal cadmium coinjection that threatens the human life and increases silver nanoparticles genotoxic risks.

  14. Sytematic Study of the Adsorption of Thiol Molecules on Noble-Metal Nanoparticles

    Science.gov (United States)

    Barron, H.; Hidalgo, F.; Fernandez-Seivane, L.; Noguez, C.; Lopez-Lozano, X.

    2012-03-01

    The study of the interaction between nanoparticles and different types of ligands has been intensively investigated in the last years due to the potential contribution of their properties to the nanotechnology device design. These properties have opened new research fields like plasmonics, with interesting applications in optics, electronics, biophysics, medicine, pharmacology and materials science. Self-assembly monolayers have been thoroughly studied at experimental and theoretical level on extended (111) gold and silver surfaces. However, nanoparticle and molecule properties after the adsorption are still not well understood due to the different factors involved in this process such as the adsorption sites, size and element type of the nanoparticle. In this work we have performed a systematic study of the adsorption of methyl-thiol molecules on Au55 and Ag55 clusters through density functional theory calculations with the SIESTA code. Different adsorption modes of the methyl-thiol molecule on Au55 and Ag55 were considered. In general, for both type of nanoparticles, the methyl-thiol molecule prefers to be adsorbed on the Bridge sites. These results provide valuable information of the structural and electronic properties of methyl-thiol passivated Au and Ag nanoparticles.

  15. A novel alternative scheme for X-ray absorption-based studies of nanoparticle growth

    Energy Technology Data Exchange (ETDEWEB)

    Meneses, Cristiano Teles [Universidade Federal de Sergipe (UFS), Itabaiana, SE (Brazil). Nucleo de Fisica; Flores, Wladimir Hernandez [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil). Dept. de Fisica; Sasaki, Jose Marcos [Universidade Federal de Pelotas, Bage, RS (Brazil). Centro de Ciencias Exatas e Tecnologicas

    2008-11-15

    In this paper we present a system developed for application in X-ray absorption spectroscopy studies using high temperatures and gases. Our focus was on an experimental setup in an in situ time-resolved study to observe the first stage of NiO nanoparticle formation carried out on the D06A DXAS beamline at LNLS. This study offers a new insight into the production of nanoparticles obtained from biopolymers with a metallic salt, and should be useful for improving the control of particle sizes in the process. (author)

  16. A targeted ultrasound contrast agent carrying gene and cell-penetrating peptide: preparation and gene transfection in vitro.

    Science.gov (United States)

    Ren, Jianli; Zhang, Ping; Tian, Ju; Zhou, Zhiyi; Liu, Xingzhao; Wang, Dong; Wang, Zhigang

    2014-09-01

    Targeted and high efficient gene delivery is a main issue in gene treatment. Taking advantage of ischemic memory target P-selectin and our previous study-synergistic effects of ultrasound-targeted microbubble destruction (UTMD) and TAT peptide on gene transfection, which were characterized by targeted aggregation and high efficient gene transfection, we set up a 'smart' gene delivery system-targeted ultrasound contrast agent (UCA) carrying gene and cell-permeable peptides (CPP). Such UCA had a strong binding force with DNA which was protected from being hydrolysed by nuclease. Moreover, synergistic effects of UTMD and TAT peptide increased gene transfection. Specifically, the UCA were reacted with an ischemic memory target P-selectin overexpressed by ischemic issues (including ischemic heart disease) and loaded with gene and CPP, which enabled targeted localization and gene delivery to ischemic cells overexpressing P-selectin. We demonstrated their targeting affinity for hypoxia human umbilical vein endothelial cell (HUVEC) and gene transfection in vitro. The results of confocal laser scanning microscopy (CLSM) showed that gene and CPP were distributed on the shell of UCA. Red fluorescence was observed on the surface of targeted UCA using immunofluorescent microscopy, which demonstrated that the antibody was successfully connected to the UCA. The targeted UCA was specifically and tightly binded to hypoxia HUVEC, while there were no or little non-targeted UCA binding around hypoxia HUVEC. 24h after transfection, gene transfection efficiency detected by FCM was higher in targeted group than non-targeted group. Overall, the targeted UCA carrying gene and CPP was prepared successfully. It had a strong target binding capacity to hypoxia HUVEC and high efficient gene transfection, which maybe provide a novel strategy for gene therapy. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Effect of Titanium dioxide nanoparticles on the flexural strength of polymethylmethacrylate: an in vitro study.

    Science.gov (United States)

    Harini, P; Mohamed, Kasim; Padmanabhan, T V

    2014-01-01

    To improve the flexural strength of polymethylmethacrylate (PMMA). To evaluate whether the incorporation of titanium dioxide nanoparticles in polymethylmethacrylate (PMMA) increases the flexural strength and to compare the different concentrations of titanium dioxide nanoparticles and its relation to flexural strength. Study was conducted in Sri Ramachandra University utilizing 40 specimens manufactured from clear heat polymerizing acrylic resin. Forty specimens of clear heat polymerizing acrylic resin of dimensions 65 Χ 10 Χ 3 mm as per ISO 1,567 standardization were fabricated and were grouped into A (CONTROL) with no titanium dioxide (TiO2) nanoparticles, B with 0.5 gms of TiO 2 nanoparticles, C with 1 gm of TiO 2 nanoparticles and D with 2.5 gms of TiO 2 nanoparticles added.The concentrations of titanium dioxide in each group were 1 wt%, 2 wt% and 5 wt%. Universal testing machine INSTRON was used to load at the center of the specimen with a cross head speed of 1.50 mm/min and a span length of 40.00 mm. ANOVA and multiple comparisons are carried out using the independent t-test. The ANOVA result shows that there is a significant difference between the groups with respect to the mean flexural strength. Highest mean flexural strength is observed in Group D, while the lowest is seen in Group A. Independent t-test revealed that there was a statistical significance between Group A and Group D (0.041) and between Group B and Group D (0.028). The results concluded that polymethylmethacrylate reinforced with different concentrations of titanium dioxide nanoparticles showed superior flexural strength than those of normal PMMA.

  18. Novel targeted siRNA-loaded hybrid nanoparticles: preparation, characterization and in vitro evaluation.

    Science.gov (United States)

    Dim, Nneka; Perepelyuk, Maryna; Gomes, Olukayode; Thangavel, Chellappagounder; Liu, Yi; Den, Robert; Lakshmikuttyamma, Ashakumary; Shoyele, Sunday A

    2015-09-26

    siRNAs have a high potential for silencing critical molecular pathways that are pathogenic. Nevertheless, their clinical application has been limited by a lack of effective and safe nanotechnology-based delivery system that allows a controlled and safe transfection to cytosol of targeted cells without the associated adverse effects. Our group recently reported a very effective and safe hybrid nanoparticle delivery system composing human IgG and poloxamer-188 for siRNA delivery to cancer cells. However, these nanoparticles need to be optimized in terms of particle size, loading capacity and encapsulation efficiency. In the present study, we explored the effects of certain production parameters on particle size, loading capacity and encapsulation efficiency. Further, to make these nanoparticles more specific in their delivery of siRNA, we conjugated anti-NTSR1-mAb to the surface of these nanoparticles to target NTSR1-overexpressing cancer cells. The mechanism of siRNA release from these antiNTSR1-mAb functionalized nanoparticles was also elucidated. It was demonstrated that the concentration of human IgG in the starting nanoprecipitation medium and the rotation speed of the magnetic stirrer influenced the encapsulation efficiency, loading capacity and the size of the nanoparticles produced. We also successfully transformed these nanoparticles into actively targeted nanoparticles by functionalizing with anti-NTSR1-mAb to specifically target NTSR1-overexpressing cancer cells, hence able to avoid undesired accumulation in normal cells. The mechanism of siRNA release from these nanoparticles was elucidated to be by Fickian diffusion. Using flow cytometry and fluorescence microscopy, we were able to confirm the active involvement of NTSR1 in the uptake of these anti-NTSR1-mAb functionalized hybrid nanoparticles by lung adenocarcinoma cells. This hybrid nanoparticle delivery system can be used as a platform technology for intracellular delivery of siRNAs to NTSR1

  19. NMR relaxation in systems with magnetic nanoparticles: a temperature study.

    Science.gov (United States)

    Issa, Bashar; Obaidat, Ihab M; Hejasee, Rola H; Qadri, Shahnaz; Haik, Yousef

    2014-03-01

    To measure and model nuclear magnetic resonance (NMR) relaxation enhancement due to the presence of gadolinium (Gd)-substituted Zn-Mn ferrite magnetic nanoparticles (MNP) at different temperatures. Relaxation rates were measured at 1.5 T using fast spin echo (FSE) sequences in samples of agarose gel doped with uncoated and polyethylene glycol (PEG)-coated Mn0.5 Zn0.5 Gd0.02 Fe1.98 O4 nanoparticles over the temperature range 8-58°C. Physical characterization of the MNPs synthesized using chemical coprecipitation included scanning (SEM) and transmission (TEM) electron microscopy, inductively coupled plasma (ICP), dynamic light scattering (DLS), and magnetometry. Relaxivity (in s(-1) mM(-1) Fe) for the uncoated and coated particles, respectively, increased as follows: from 2.5 to 3.2 and 0.4 to 0.7 for T1, while for T2 it increased from 162.3 to 253.7 and 59.7 to 82.2 over the temperature range 8-58°C. T2 data were fitted to the echo limited motional regime using one fitting parameter that reflects the degree of agglomeration of particles into a cluster. This parameter was found to increase linearly with temperature and was larger for the PEG-coated particles than the uncoated ones. The increase of 1/T2 with temperature is modeled successfully using echo limited motional regime where both diffusion of the protons and nanoparticle cluster size increase with temperature. Both transverse and longitudinal relaxation efficiencies are reduced by PEG coating at all temperatures. If prediction of relaxation rates under different particle concentrations and operating temperatures is possible then the use of MNP in temperature monitoring and hyperthermia applications may be achieved. Copyright © 2013 Wiley Periodicals, Inc.

  20. Theoretical study of ferroelectric nanoparticles using phase reconstructed electron microscopy

    DEFF Research Database (Denmark)

    Phatak, C.; Petford-Long, A. K.; Beleggia, Marco

    2014-01-01

    Ferroelectric nanostructures are important for a variety of applications in electronic and electro-optical devices, including nonvolatile memories and thin-film capacitors. These applications involve stability and switching of polarization using external stimuli, such as electric fields. We present...... a theoretical model describing how the shape of a nanoparticle affects its polarization in the absence of screening charges, and quantify the electron-optical phase shift for detecting ferroelectric signals with phase-sensitive techniques in a transmission electron microscope. We provide an example phase shift...

  1. Theoretical study of the amphoteric oxide nanoparticle surface charge during multi-particle interactions in aqueous solutions

    Science.gov (United States)

    Alfimov, A. V.; Aryslanova, E. M.; Chivilikhin, S. A.

    2015-11-01

    Nanoparticle surface charge plays an important role in many biological applications. In this study, an analytical surface charging model for the amphoteric oxide nanoparticles has been presented. The model accounts for the particle's electric double layer self-action on the charging process and the charge regulation during multi-particle interactions in aqueous solutions. The employment of the model allows to explicitly describe the nanoparticle agglomeration process and the accompanying agglomerate surface charge variation.

  2. A Study of Efficiency of Zero-valent Iron Nanoparticles in Degradation of Trichlorethylene from Aqueous Solutions

    Directory of Open Access Journals (Sweden)

    Samaneh Dehghan

    2016-12-01

    mg/l, and contact time= 86 min. The results of kinetic studies revealed that TCE degradation by nZVI follows first-order kinetic model. Conclusion: It is conclude that zero-valent iron nanoparticles have a good efficiency in the degradation of TCE. On the other hand, separation of these nanoparticles is simple due to its magnetism properties, which can improve the use of these nanoparticles

  3. Biocompatibility study of protein capped and uncapped silver nanoparticles on human hemoglobin

    International Nuclear Information System (INIS)

    Bhunia, Amit Kumar; Saha, Satyajit; Samanta, Pijus Kanti; Aich, Debasish; Kamilya, Tapanendu

    2015-01-01

    The interactions of human hemoglobin with protein capped silver nanoparticles and bare silver nanoparticles were studied to understand fundamental perspectives about the biocompatibility of protein capped silver nanoparticles compared with bare silver nanoparticles. Bare silver (Ag) nanoparticles (NPs) were prepared by the chemical reduction method. High resolution transmission electron microscopy (HRTEM) analysis along with absorption at ∼390 nm indicated the formation of bare Ag NPs. Protein coated Ag NPs were prepared by a green synthesis method. Absorption at ∼440 nm along with ∼280 nm indicated the formation of protein coated Ag NPs. The biocompatibility of the above mentioned Ag NPs was studied by interaction with human hemoglobin (Hb) protein. In presence of bare Ag NPs, the Soret band of Hb was red shifted. This revealed the distortion of iron from the heme pockets of Hb. Also, the fluorescence peak of Hb was quenched and red shifted which indicated that Hb became unfolded in the presence of bare Ag NPs. No red shift of the absorption of Soret, along with no shift and quenching of the fluorescence peak of Hb were observed in the presence of protein coated Ag NPs. A hemolysis assay suggested that protein coated Ag NPs were more biocompatible than bare one. (paper)

  4. Graphene substrate-mediated catalytic performance enhancement of Ru nanoparticles: A first-principles study

    KAUST Repository

    Liu, Xin

    2012-01-01

    The structural, energetic and magnetic properties of Ru nanoparticles deposited on pristine and defective graphene have been thoroughly studied by first-principles based calculations. The calculated binding energy of a Ru 13 nanoparticle on a single vacancy graphene is as high as -7.41 eV, owing to the hybridization between the dsp states of the Ru particles with the sp 2 dangling bonds at the defect sites. Doping the defective graphene with boron would further increase the binding energy to -7.52 eV. The strong interaction results in the averaged d-band center of the deposited Ru nanoparticle being upshifted toward the Fermi level from -1.41 eV to -1.10 eV. Further study reveals that the performance of the nanocomposites against hydrogen, oxygen and carbon monoxide adsorption is correlated to the shift of the d-band center of the nanoparticle. Thus, Ru nanoparticles deposited on defective graphene are expected to exhibit both high stability against sintering and superior catalytic performance in hydrogenation, oxygen reduction reaction and hydrogen evolution reaction. © 2012 The Royal Society of Chemistry.

  5. Biocompatibility study of protein capped and uncapped silver nanoparticles on human hemoglobin

    Science.gov (United States)

    Bhunia, Amit Kumar; Kanti Samanta, Pijus; Aich, Debasish; Saha, Satyajit; Kamilya, Tapanendu

    2015-06-01

    The interactions of human hemoglobin with protein capped silver nanoparticles and bare silver nanoparticles were studied to understand fundamental perspectives about the biocompatibility of protein capped silver nanoparticles compared with bare silver nanoparticles. Bare silver (Ag) nanoparticles (NPs) were prepared by the chemical reduction method. High resolution transmission electron microscopy (HRTEM) analysis along with absorption at ~390 nm indicated the formation of bare Ag NPs. Protein coated Ag NPs were prepared by a green synthesis method. Absorption at ~440 nm along with ~280 nm indicated the formation of protein coated Ag NPs. The biocompatibility of the above mentioned Ag NPs was studied by interaction with human hemoglobin (Hb) protein. In presence of bare Ag NPs, the Soret band of Hb was red shifted. This revealed the distortion of iron from the heme pockets of Hb. Also, the fluorescence peak of Hb was quenched and red shifted which indicated that Hb became unfolded in the presence of bare Ag NPs. No red shift of the absorption of Soret, along with no shift and quenching of the fluorescence peak of Hb were observed in the presence of protein coated Ag NPs. A hemolysis assay suggested that protein coated Ag NPs were more biocompatible than bare one.

  6. Mathematical study of probe arrangement and nanoparticle injection effects on heat transfer during cryosurgery.

    Science.gov (United States)

    Mirkhalili, Seyyed Mostafa; Ramazani S A, Ahmad; Nazemidashtarjandi, Saeed

    2015-11-01

    Blood vessels, especially large vessels have a greater thermal effect on freezing tissue during cryosurgery. Vascular networks act as heat sources in tissue, and cause failure in cryosurgery and reappearance of cancer. The aim of this study is to numerically simulate the effect of probe location and multiprobe on heat transfer distribution. Furthermore, the effect of nanoparticles injection is studied. It is shown that the small probes location near large blood vessels could help to reduce the necessary time for tissue freezing. Nanoparticles injection shows that the thermal effect of blood vessel in tissue is improved. Using Au, Ag and diamond nanoparticles have the most growth of ice ball during cryosurgery. However, polytetrafluoroethylene (PTFE) nanoparticle can be used to protect normal tissue around tumor cell due to its influence on reducing heat transfer in tissue. Introduction of Au, Ag and diamond nanoparticles combined with multicryoprobe in this model causes reduction of tissue average temperature about 50% compared to the one probe. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Synthesis, Antibacterial and Thermal Studies of Cellulose Nanocrystal Stabilized ZnO-Ag Heterostructure Nanoparticles

    Directory of Open Access Journals (Sweden)

    Mohd Zobir Hussein

    2013-05-01

    Full Text Available Synthesis of ZnO-Ag heterostructure nanoparticles was carried out by a precipitation method with cellulose nanocrystals (CNCs as a stabilizer for antimicrobial and thermal studies. ZnO-Ag nanoparticles were obtained from various weight percentages of added AgNO3 relative to Zn precursors for evaluating the best composition with enhanced functional properties. The ZnO-Ag/CNCs samples were characterized systematically by TEM, XRD, UV, TGA and DTG. From the TEM studies we observed that ZnO-Ag heterostructure nanoparticles have spherical shapes with size diameters in a 9–35 nm range. The antibacterial activities of samples were assessed against the bacterial species Salmonella choleraesuis and Staphylococcus aureus. The CNC-stabilized ZnO-Ag exhibited greater bactericidal activity compared to cellulose-free ZnO-Ag heterostructure nanoparticles of the same particle size. The incorporation of ZnO-Ag hetreostructure nanoparticles significantly increased the thermal stability of cellulose nanocrystals.

  8. Characterizing the structure of lipodisq nanoparticles for membrane protein spectroscopic studies.

    Science.gov (United States)

    Zhang, Rongfu; Sahu, Indra D; Liu, Lishan; Osatuke, Anna; Comer, Raven G; Dabney-Smith, Carole; Lorigan, Gary A

    2015-01-01

    Membrane protein spectroscopic studies are challenging due to the difficulty introduced in preparing homogenous and functional hydrophobic proteins incorporated into a lipid bilayer system. Traditional membrane mimics such as micelles or liposomes have proved to be powerful in solubilizing membrane proteins for biophysical studies, however, several drawbacks have limited their applications. Recently, a nanosized complex termed lipodisq nanoparticles was utilized as an alternative membrane mimic to overcome these caveats by providing a homogeneous lipid bilayer environment. Despite all the benefits that lipodisq nanoparticles could provide to enhance the biophysical studies of membrane proteins, structural characterization in different lipid compositions that closely mimic the native membrane environment is still lacking. In this study, the formation of lipodisq nanoparticles using different weight ratios of POPC/POPG lipids to SMA polymers was characterized via solid-state nuclear magnetic resonance (SSNMR) spectroscopy and dynamic light scattering (DLS). A critical weight ratio of (1/1.25) for the complete solubilization of POPC/POPG vesicles has been observed and POPC/POPG vesicles turned clear instantaneously upon the addition of the SMA polymer. The size of lipodisq nanoparticles formed from POPC/POPG lipids at this weight ratio of (1/1.25) was found to be about 30 nm in radius. We also showed that upon the complete solubilization of POPC/POPG vesicles by SMA polymers, the average size of the lipodisq nanoparticles is weight ratio dependent, when more SMA polymers were introduced, smaller lipodisq nanoparticles were obtained. The results of this study will be helpful for a variety of biophysical experiments when specific size of lipid disc is required. Further, this study will provide a proper path for researchers working on membrane proteins to obtain pertinent structure and dynamic information in a physiologically relevant membrane mimetic environment

  9. [Cashmere goat bacterial artificial chromosome recombination and cell transfection system].

    Science.gov (United States)

    Huang, Tian; Cao, Zhongyang; Yang, Yaohui; Cao, Gengsheng

    2016-03-01

    The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.

  10. Evolution of ZnS Nanoparticles via Facile CTAB Aqueous Micellar Solution Route: A Study on Controlling Parameters

    Directory of Open Access Journals (Sweden)

    Gradzielski Michael

    2008-01-01

    Full Text Available Abstract Synthesis of semiconductor nanoparticles with new photophysical properties is an area of special interest. Here, we report synthesis of ZnS nanoparticles in aqueous micellar solution of Cetyltrimethylammonium bromide (CTAB. The size of ZnS nanodispersions in aqueous micellar solution has been calculated using UV-vis spectroscopy, XRD, SAXS, and TEM measurements. The nanoparticles are found to be polydispersed in the size range 6–15 nm. Surface passivation by surfactant molecules has been studied using FTIR and fluorescence spectroscopy. The nanoparticles have been better stabilized using CTAB concentration above 1 mM. Furthermore, room temperature absorption and fluorescence emission of powdered ZnS nanoparticles after redispersion in water have also been investigated and compared with that in aqueous micellar solution. Time-dependent absorption behavior reveals that the formation of ZnS nanoparticles depends on CTAB concentration and was complete within 25 min.

  11. Structural studies on drop-cast film based on functionalized gold nanoparticles network: The effect of thermal treatment

    International Nuclear Information System (INIS)

    Fontana, Laura; Fratoddi, Ilaria; Venditti, Iole; Ksenzov, Dmitriy; Russo, Maria Vittoria; Grigorian, Souren

    2016-01-01

    Graphical abstract: - Highlights: • Gold nanoparticles functionalized by π-conjugated dithiol. • In situ annealing of a gold nanoparticles network. • Structural reorganization of a gold nanoparticles network before and after thermal treatment. - Abstract: In the present work the role of the thermal treatment on the reorganization of gold nanoparticles (AuNPs) functionalized with a π-conjugated dithiol ligand, namely 9,9-didodecyl-2,7-bis-thiofluorene, is studied by grazing incidence X-ray diffraction technique. For a detailed investigation of the structural changes and reorganization occurring in the AuNPs network and of the monitoring of complex interactions between nanoparticles, the line profiles are analyzed in out-of-plane and in-plane directions. The obtained data support the idea of the formation of a uniform network of nanoparticles that after annealing are extended from hexagonal to cubic arrangement.

  12. Study of energy transfer between molecules placed in the vicinity of a bimetal composite nanoparticle

    Energy Technology Data Exchange (ETDEWEB)

    Daneshfar, Nader, E-mail: ndaneshfar@gmail.com, E-mail: ndaneshfar@razi.ac.ir [Department of Physics, Razi University, Kermanshah (Iran, Islamic Republic of)

    2015-10-15

    In this study, the problem of energy transfer between two molecules near a bimetallic composite nanoparticle is investigated. The influence of the interaction between metal particles on the intermolecular energy is studied, because when two metal nanoparticles are placed close to each other, their plasmons coupling giving rise to new features. On the other hand, we discuss the transfer of resonance energy between donor and acceptor molecules (a single donor and a single acceptor) in the presence of a nanocomposite containing gold and silver nanoparticles based on the Maxwell-Garnett effective medium theory and within the quasistatic limit. We show that the interaction energy strongly depends on the particle size, the filling factor of metal particles, the intermolecular distance (the distance between the donor and acceptor molecules), and the dielectric constant of host matrix.

  13. The Synthesis, Characterization and Catalytic Reaction Studies of Monodisperse Platinum Nanoparticles in Mesoporous Oxide Materials

    Energy Technology Data Exchange (ETDEWEB)

    Rioux, Robert M. [Univ. of California, Berkeley, CA (United States)

    2006-01-01

    A catalyst design program was implemented in which Pt nanoparticles, either of monodisperse size and/or shape were synthesized, characterized and studied in a number of hydrocarbon conversion reactions. The novel preparation of these materials enables exquisite control over their physical and chemical properties that could be controlled (and therefore rationally tuned) during synthesis. The ability to synthesize rather than prepare catalysts followed by thorough characterization enable accurate structure-function relationships to be elucidated. This thesis emphasizes all three aspects of catalyst design: synthesis, characterization and reactivity studies. The precise control of metal nanoparticle size, surface structure and composition may enable the development of highly active and selective heterogeneous catalysts.

  14. Spectroelectrochemical and morphological studies of the ageing of silver nanoparticles embedded in ultra-thin perfluorinated sputter deposited films

    Energy Technology Data Exchange (ETDEWEB)

    Ebbert, C., E-mail: ebbert@tc.upb.de [University of Paderborn, Faculty of Natural Science, Technical and Macromolecular Chemistry, Warburger Str. 100, 33098 Paderborn (Germany); Alissawi, N. [Institute for Materials Science, Christian-Albrechts University at Kiel, Kaiserstr. 2, 24143 Kiel (Germany); Somsen, C.; Eggeler, G. [Institute of Materials, Department of Mechanical Engineering, Ruhr-University Bochum, Universitaetst. 150, 44780 Bochum (Germany); Strunskus, T.; Faupel, F. [Institute for Materials Science, Christian-Albrechts University at Kiel, Kaiserstr. 2, 24143 Kiel (Germany); Grundmeier, G. [University of Paderborn, Faculty of Natural Science, Technical and Macromolecular Chemistry, Warburger Str. 100, 33098 Paderborn (Germany)

    2014-11-28

    This paper focuses on the investigation of the ageing behaviour of silver nanoparticle containing polytetrafluoroethylene thin films during exposure to phosphate buffer solution (pH = 7.5). In order to investigate the effect of the electrical connection between the silver nanoparticles via a conductive substrate, two kinds of composite films were compared. One model where the nanoparticles are directly deposited on an inert conducting substrate and then covered by an ultra-thin polytetrafluoroethylene like film. In the second case a polytetrafluoroethylene/silver nanoparticle/polytetrafluoroethylene sandwich film was prepared on the same substrate to prevent electrical connection of the silver nanoparticles. Degradation was followed in-situ by means of the combination of ultraviolet–visible spectroscopy and electrochemical impedance spectroscopy. In the case of electrically connected nanoparticles electrochemical Ostwald ripening took place, while this process was not observed for the insulated nanoparticles. The electrochemical impedance spectroscopy studies allowed for the parallel study of the correlated loss of barrier properties. Transmission electron microscopy images of both composite films confirmed the results obtained by means of the in situ electrochemical ultraviolet–visible studies. - Highlights: • Nanoparticle in polymer films could be analysed by a spectroelectrochemical approach. • Transmission electron microscopy analysis proved an Ostwald-ripening process. • Embedding of the silver nanoparticles inhibits the Ostwald-ripening process.

  15. Twenty-eight-day inhalation toxicity study of silver nanoparticles in Sprague-Dawley rats.

    Science.gov (United States)

    Ji, Jun Ho; Jung, Jae Hee; Kim, Sang Soo; Yoon, Jin-Uk; Park, Jung Duck; Choi, Byung Sun; Chung, Yong Hyun; Kwon, Il Hoon; Jeong, Jayoung; Han, Beom Seok; Shin, Jae Hyeg; Sung, Jae Hyuck; Song, Kyung Seuk; Yu, Il Je

    2007-08-01

    The antibacterial effect of silver nanoparticles has resulted in their extensive application in health, electronic, and home products. Thus, the exposed population continues to increase as the applications expand. Although previous studies on silver dust, fumes, and silver compounds have revealed some insights, little is yet known about the toxicity of nano-sized silver particles, where the size and surface area are recognized as important determinants for toxicity. Thus, the inhalation toxicity of silver nanoparticles is of particular concern to ensure the health of workers and consumers. However, the dispersion of inhalable ambient nano-sized particles has been an obstacle in evaluating the effect of the inhalation of nano-sized particles on the respiratory system. Accordingly, the present study used a device that generates silver nanoparticles by evaporation/condensation using a small ceramic heater. As such, the generator was able to distribute the desired concentrations of silver nanoparticles to chambers containing experimental animals. The concentrations and distribution of the nanoparticles with respect to size were also measured directly using a differential mobility analyzer and ultrafine condensation particle counter. Therefore, the inhalation toxicity of silver nanoparticles was tested over a period of 28 days. Eight-week-old rats, weighing about 283 g for the males and 192 g for the females, were divided into 4 groups (10 rats in each group): a fresh-air control, a low-dose group (1.73 x 10(4)/cm3), a middle-dose group (1.27 x 10(5)/cm3), and a high-dose group (1.32 x 10(6) particles/cm3, 61 microg/m3). The animals were exposed to the silver nanoparticles for 6 h/day, 5 days/wk, for a total of 4 wk. The male and female rats did not show any significant changes in body weight relative to the concentration of silver nanoparticles during the 28-day experiment. Plus, there were no significant changes in the hematology and blood biochemical values in either

  16. Synthesis and Stability of Iron Nanoparticles for Lunar Environment Studies

    Science.gov (United States)

    Hung, Ching-cheh; McNatt, Jeremiah

    2009-01-01

    Simulant of lunar dust is needed when researching the lunar environment. However, unlike the true lunar dust, today s simulants do not contain nanophase iron. Two different processes have been developed to fabricate nanophase iron to be used as part of the lunar dust simulant: (1) Sequentially treating a mixture of ferric chloride, fluorinated carbon, and soda lime glass beads at about 300 C in nitrogen, at room temperature in air, and then at 1050 C in nitrogen. The product includes glass beads that are grey in color, can be attracted by a magnet, and contain alpha-iron nanoparticles (which seem to slowly lose their lattice structure in ambient air during a period of 12 months). This product may have some similarity to the lunar glassy regolith that contains Fe(sup 0). (2) Heating a mixture of carbon black and a lunar simulant (a mixed metal oxide that includes iron oxide) at 1050 C in nitrogen. This process simulates lunar dust reaction to the carbon in a micrometeorite at the time of impact. The product contains a chemically modified simulant that can be attracted by a magnet and has a surface layer whose iron concentration increased during the reaction. The iron was found to be alpha-iron and Fe3O4 nanoparticles, which appear to grow after the fabrication process, but stabilizes after 6 months of ambient air storage.

  17. Theoretical studies of acrolein hydrogenation on Au20 nanoparticle

    Science.gov (United States)

    Li, Zhe; Chen, Zhao-Xu; He, Xiang; Kang, Guo-Jun

    2010-05-01

    Gold nanoparticles play a key role in catalytic processes. We investigated the kinetics of stepwise hydrogenation of acrolein on Au20 cluster model and compared with that on Au(110) surface. The rate-limiting step barrier of CC reduction is about 0.5 eV higher than that of CO hydrogenation on Au(110) surface. On Au20 nanoparticle, however, the energy barrier of the rate-determining step for CC hydrogenation turns out to be slightly lower than the value for the CO reduction. The selectivity difference on the two substrate models are attributed to different adsorption modes of acrolein: via the CC on Au20, compared to through both CC and CO on Au(110). The preference switch implies that the predicted selectivity of competitive hydrogenation depends on substrate model sensitively, and particles with more low-coordinated Au atoms than flat surfaces are favorable for CC hydrogenation, which is in agreement with experimental result.

  18. Studies on Characterization, Optical Absorption, and Photoluminescence of Yttrium Doped ZnS Nanoparticles

    Directory of Open Access Journals (Sweden)

    Ranganaik Viswanath

    2014-01-01

    Full Text Available Pure ZnS and ZnS:Y nanoparticles were synthesized by a chemical coprecipitation route using EDTA-ethylenediamine as a stabilizing agent. X-ray diffraction (XRD, high resolution transmission electron microscopy (HRTEM, field emission scanning electron microscopy (FE-SEM, Fourier transform infrared spectrometry (FTIR, thermogravimetric-differential scanning calorimetry (TG-DSC, and UV-visible and photoluminescence (PL spectroscopy were employed to characterize the as-synthesized ZnS and ZnS:Y nanoparticles, respectively. XRD and TEM studies show the formation of cubic ZnS:Y particles with an average size of ~4.5 nm. The doping did not alter the phase of the zinc sulphide, as a result the sample showed cubic zincblende structure. The UV-visible spectra of ZnS and ZnS:Y nanoparticles showed a band gap energy value, 3.85 eV and 3.73 eV, which corresponds to a semiconductor material. A luminescence characteristics such as strong and stable visible-light emissions in the orange region alone with the blue emission peaks were observed for doped ZnS nanoparticles at room temperature. The PL intensity of orange emission peak was found to be increased with an increase in yttrium ions concentration by suppressing blue emission peaks. These results strongly propose that yttrium doped zinc sulphide nanoparticles form a new class of luminescent material.

  19. The Study of growth and coagulation of titania nanoparticles by chemical vapor synthesis

    International Nuclear Information System (INIS)

    Rahiminezhad-Soltani, M.; Saberyan, K.; Shahri, F.; Simchi, A.

    2010-01-01

    Chemical Vapor Synthesis route was used for synthesis of titanium dioxide (TiO 2 ) nanoparticles in hot-walled reactor at 800 d egree C using TiCl 4 as precursor. The effect of processing parameters e.g., temperature and amount of precursor on phase structure, size, purity, coagulation and agglomeration of nanoparticles were investigated in this respect. Also, the H 2 O effects on the size, crystallinity, phase transformation and purity of nanoparticles were studied. Comprehensive experimental observations were confirmed by transmission electron microscopy, X-ray diffraction analysis and thermal gravimetric-differential thermal analysis results. The obtained results showed that by increasing the precursor amount and temperature, no phase transformation can be observed but the size, coagulation and agglomeration of titania nanoparticles increase. Also, the results showed that by introducing water vapor, the average particle sizes decrease saliently and no phase transformation and impurity were observed. Titanium dioxide nanoparticles can be used for synthesis of nano fluids. Nano fluids (nano-TiO 2 +water) as a cooling agent can be used for the enhanced economy and safety of the nuclear reactors.

  20. Toxicity Study of Silver Nanoparticles Synthesized from Suaeda monoica on Hep-2 Cell Line.

    Science.gov (United States)

    Satyavani, Kaliyamurthi; Gurudeeban, Selvaraj; Ramanathan, Thiruganasambandam; Balasubramanian, Thangavel

    2012-01-01

    Recently there has been fabulous excitement in the nano-biotechnological area for the study of nanoparticles synthesis using some natural biological system, which has led the growth advanced nanomaterials. This intention made us to assess the biologically synthesized silver nanoparticles from the leaf of Suaeda monoica (S.monoica) using 1 mM silver nitrate. The leaf extract of S.monoica incubated with 1 mM silver nitrate solution and characterized by UV- spectrometer and AFM. The effect of synthesized silver nanoparticles on Human Epidermoid Larynx Carcinoma cell line was evaluated by the MTT colorimetric technique. As a result we observed gradual change in the colour of extract from greenish to brown. The synthesized silver nanoparticles confirmed by UV at 430 nm and spherical shape identified in the range of 31 nm under AFM. The effect of silver nanoparticles on Human Epidermoid Larynx Carcinoma cell line exhibits a dose-dependent toxicity for the cell tested and the viability of Hep-2 cells decreased to 50 % (IC(50)) at the concentration of 500 nM. Further findings will be determined the exact mechanisms of this cost effective Nano-treatments.

  1. Studies of aggregated nanoparticles steering during magnetic-guided drug delivery in the blood vessels

    Energy Technology Data Exchange (ETDEWEB)

    Hoshiar, Ali Kafash [School of Mechanical and Aerospace Engineering and ReCAPT, Gyeongsang National University, Jinju 660-701 (Korea, Republic of); Faculty of Industrial and Mechanical Engineering, Islamic Azad University, Qazvin Branch, Qazvin (Iran, Islamic Republic of); Le, Tuan-Anh [School of Mechanical and Aerospace Engineering and ReCAPT, Gyeongsang National University, Jinju 660-701 (Korea, Republic of); Amin, Faiz Ul [Department of Biology and Applied Life Science, Gyeongsang National University, Jinju 660-701 (Korea, Republic of); Kim, Myeong Ok, E-mail: mokim@gnu.ac.kr [Department of Biology and Applied Life Science, Gyeongsang National University, Jinju 660-701 (Korea, Republic of); Yoon, Jungwon, E-mail: jwyoon@gnu.ac.kr [School of Mechanical and Aerospace Engineering and ReCAPT, Gyeongsang National University, Jinju 660-701 (Korea, Republic of)

    2017-04-01

    Magnetic-guided targeted drug delivery (TDD) systems can enhance the treatment of diverse diseases. Despite the potential and promising results of nanoparticles, aggregation prevents precise particle guidance in the vasculature. In this study, we developed a simulation platform to investigate aggregation during steering of nanoparticles using a magnetic field function. The magnetic field function (MFF) comprises a positive and negative pulsed magnetic field generated by electromagnetic coils, which prevents adherence of particles to the vessel wall during magnetic guidance. A commonly used Y-shaped vessel was simulated and the performance of the MFF analyzed; the experimental data were in agreement with the simulation results. Moreover, the effects of various parameters on magnetic guidance were evaluated and the most influential identified. The simulation results presented herein will facilitate more precise guidance of nanoparticles in vivo. - Highlights: • We developed a simulation platform to investigate aggregation of nanoparticles. • The influential parameters for magnetic steering of a Y-shaped vessel were identified. • The proposed platform will facilitate more precise guidance of nanoparticles in vivo.

  2. Green synthesis of silver nanoparticles using Terminalia chebula extract at room temperature and their antimicrobial studies

    Science.gov (United States)

    Mohan Kumar, Kesarla; Sinha, Madhulika; Mandal, Badal Kumar; Ghosh, Asit Ranjan; Siva Kumar, Koppala; Sreedhara Reddy, Pamanji

    2012-06-01

    A green rapid biogenic synthesis of silver nanoparticles (Ag NPs) using Terminalia chebula (T. chebula) aqueous extract was demonstrated in this present study. The formation of silver nanoparticles was confirmed by Surface Plasmon Resonance (SPR) at 452 nm using UV-visible spectrophotometer. The reduction of silver ions to silver nanoparticles by T. chebula extract was completed within 20 min which was evidenced potentiometrically. Synthesised nanoparticles were characterised using UV-vis spectroscopy, Fourier transformed infrared spectroscopy (FT-IR), powder X-ray diffraction (XRD), transmission electron microscopy (TEM) and atomic force microscopy (AFM). The hydrolysable tannins such as di/tri-galloyl-glucose present in the extract were hydrolyzed to gallic acid and glucose that served as reductant while oxidised polyphenols acted as stabilizers. In addition, it showed good antimicrobial activity towards both Gram-positive bacteria (S. aureus ATCC 25923) and Gram-negative bacteria (E. coli ATCC 25922). Industrially it may be a smart option for the preparation of silver nanoparticles.

  3. Studies on the Genotoxicity Behavior of Silver Nanoparticles in the Presence of Heavy Metal Cadmium Chloride in Mice

    OpenAIRE

    Mohamed, Hanan Ramadan Hamad

    2016-01-01

    Incredible rapid growth in the nanoparticles applications and development increases the daily human exposure to them but humans are exposed to many other pollutants in addition to nanoparticles that forced us to evaluate the effect of heavy metal cadmium chloride (CdCl2) coinjection on silver nanoparticles induced genotoxic risk in this study. Mice were injected into the abdominal cavity with single dose of Ag nanoparticles (20, 41, and 82 mg/kg) or CdCl2 (1.5 mg/kg) either separately or toge...

  4. Endothelial cell transfection of ex vivo arteries

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Alexander Lohman, Adam Straub & Brant Isakson ### Abstract The vascular endothelium plays an essential role in regulating blood vessel tone, blood flow and blood pressure. Current vascular model systems for examination of endothelial cell biology and blood vessel physiology and pathology rely on cell culture and the generation of genetically modified animals. While these systems are advantageous for studying the endothelium, many cell culture models omit the contribution of o...

  5. Study of the Carbonization and Graphitization of Coal Tar Pitch Modified with SiC Nanoparticles

    Directory of Open Access Journals (Sweden)

    Maciej Gubernat

    2017-01-01

    Full Text Available Silicon carbide nanoparticles (nSiC have been used to modify coal tar pitch (CTP as a carbon binder. The influence of ceramic nanoparticles on the structure and microstructure was studied. The structure of CTP-based carbon residue with various nSiC contents was analyzed by using SEM with EDAX, Raman spectroscopy, and X-ray diffraction. The effect of ceramic nanofiller on the crystallite sizes (Lc, La and the c-axis spacing (d002 in carbonized samples after heating from 1000 to 2800°C was analyzed. Ceramic nanofillers inhibit structural changes in carbonized samples heated to 1000°C. After heating CTP with nSiC above 2000°C, the carbon samples contained two carbon components differing in structural ordering. Ceramic nanoparticles increase carbon crystallite growth, while their impact on the c-axis spacing is low.

  6. Study of thermal diffusivity of nanofluids with bimetallic nanoparticles with Au(core)/Ag(shell) structure

    International Nuclear Information System (INIS)

    Gutierrez Fuentes, R.; Pescador Rojas, J.A.; Jimenez-Perez, J.L.; Sanchez Ramirez, J.F.; Cruz-Orea, A.; Mendoza-Alvarez, J.G.

    2008-01-01

    The thermal diffusivity of Au/Ag nanoparticles with core/shell structure, at different compositions (Au/Ag = 3/1, 1/1, 1/3, 1/6), was measured by using the mismatched mode of the dual-beam thermal lens (TL) technique. This study determines the effect of the bimetallic composition on the thermal diffusivity of the nanofluids. In these results we find a lineal increment of the nanofluid it thermal diffusivity when the Ag shell thickness is increased. Our results show that the nanoparticle structure is an important parameter to improve the heat transport in composites and nanofluids. These results could have importance for applications in therapies and photothermal deliberation of drugs. Complementary measurements with UV-vis spectroscopy and TEM, were used to characterize the Au(core)/Ag(shell) nanoparticles

  7. Structural and Thermal Studies of ZnS and CdS Nanoparticles in Polymer Matrices

    Directory of Open Access Journals (Sweden)

    Jejenija Osuntokun

    2016-01-01

    Full Text Available We report the synthesis and structural studies of ZnS and CdS nanoparticles in polyvinylpyrrolidone (PVP, poly(vinyl alcohol (PVA, and poly(methyl methacrylate (PMMA matrices. The metal sulfides/polymer nanocomposites were characterized by X-ray diffraction (XRD, Fourier transform infrared spectroscopy, electronic spectroscopy (UV-Vis, transmission electron microscopy (TEM, and thermogravimetric analysis (TGA. The particle sizes as calculated from the absorption spectra were in agreement with the results obtained from TEM and XRD data. They showed metal sulfides nanoparticles in the polymers matrices with average crystallite sizes of 1.5–6.9 nm. The TGA results indicate that incorporation of the nanoparticles significantly altered the thermal properties of the respective polymers with ZnS/PVA and CdS/PVA nanocomposites displaying higher thermal stability than the other polymer nanocomposites.

  8. [Terahertz-band study on surface enhanced Raman scattering of nanoparticle].

    Science.gov (United States)

    Wu, Yu-Deng; Ren, Guang-Jun; Hao, Yun; Yao, Jian-Quan

    2013-05-01

    Study on surface-enhanced Raman scattering in the terahertz-band proved in that the terahertz-band Raman enhancement also exists. By studing principles of electromagnetic enhancement of surface-enhanced Raman scattering, using the finite difference time-domain method, the electromagnetic enhancement of surface enhanced Raman scattering of nano-particles irradiated by terahertz-wave was simulated, and the enhancement effect of terahertz waves was analyzed. Simulation experiments show that using finite-difference time-domain method could obtain effectively accurate simulation result of nano-particle scattering, proving that for terahertz waves, surface-enhanced effects on the surface of the nano-particle also exist. The results for surface enhanced Raman scattering extended from the visible and infrared to terahertz-band, and provide a basis for application of the combination of surface-enhanced Raman scattering and terahertz-wave.

  9. Field emission studies of silver nanoparticles synthesized by electron cyclotron resonance plasma

    International Nuclear Information System (INIS)

    Purohit, Vishwas; Mazumder, Baishakhi; Bhise, A.B.; Poddar, Pankaj; Joag, D.S.; Bhoraskar, S.V.

    2011-01-01

    Field emission has been studied for silver nanoparticles (25-200 nm), deposited within a cylindrical silver target in an electron cyclotron resonance (ECR) plasma. Particle size distribution was controlled by optimum biasing voltages between the chamber and the target. Presence of non-oxidized silver was confirmed from the X-Ray diffraction analysis; however, thin protective layer of oxide was identified from the selective area electron diffraction pattern obtained with transmission electron microscopy. The silver nanoparticles were seen to exhibit hilly pointed like structures when viewed under the atomic force microscopy (AFM). The emissive properties of these particles were investigated by field emission microscopy. It is found that this technique of deposition is ideal for formation of nanoparticles films on different substrate geometries with size controllability as well as its application to emission devices.

  10. Integrating sphere study of the optical properties of aluminum nanoparticles in tetranitropentaerytrite

    Science.gov (United States)

    Aduev, B. P.; Nurmukhametov, D. R.; Belokurov, G. M.; Zvekov, A. A.; Kalenskii, A. V.; Nikitin, A. P.; Liskov, I. Yu.

    2014-09-01

    The dependences of the transmission coefficients and the sum of the transmission and absorption coefficients for light with a wavelength of 643 nm in compressed pentaerythritol tetranitrate (PETN) samples containing aluminum nanoinclusions (average radius of 50 nm) on the pellet thickness and the mass fraction of aluminum nanoparticles in it are studied with an integrating sphere. The light absorption and scattering in this system is simulated using the Mie theory and a radiative transfer equation. The depth profile of absorbed energy in a sample is shown to approximately obey the Bouguer-Beer law. The effective cross section of radiation absorption by aluminum nanoparticles, which takes into account both light absorption and scattering by an ensemble of nanoparticles, exceeds the geometrical cross section.

  11. Electrical sintering of silver nanoparticle ink studied by in-situ TEM probing.

    Directory of Open Access Journals (Sweden)

    Magnus Hummelgård

    Full Text Available Metallic nanoparticle inks are used for printed electronics, but to reach acceptable conductivity the structures need to be sintered, usually using a furnace. Recently, sintering by direct resistive heating has been demonstrated. For a microscopic understanding of this Joule heating sintering method, we studied the entire process in real time inside a transmission electron microscope equipped with a movable electrical probe. We found an onset of Joule heating induced sintering and coalescence of nanoparticles at power levels of 0.1-10 mW/μm³. In addition, a carbonization of the organic shells that stabilize the nanoparticles were found, with a conductivity of 4 10⁵ Sm⁻¹.

  12. Green synthesis of silver nanoparticles by Ricinus communis var. carmencita leaf extract and its antibacterial study

    Science.gov (United States)

    Ojha, Sunita; Sett, Arghya; Bora, Utpal

    2017-09-01

    In this study, we report synthesis of silver nanoparticles (RcAgNPs) from silver nitrate solution using methanolic leaf extract of Ricinus communis var. carmencita. The polyphenols present in the leaves reduce Ag++ ions to Ag0 followed by a color change. Silver nanoparticle formation was ensured by surface plasmon resonance between 400 nm to 500 nm. Crystallinity of the synthesized nanoparticles was confirmed by UHRTEM, SAED and XRD analysis. The capping of phytochemicals and thermal stability of RcAgNPs were assessed by FTIR spectra and TGA analysis, respectively. It also showed antibacterial activity against both gram positive and gram negative strains. RcAgNPs were non-toxic against normal cell line (mouse fibroblast cell line L929) at lower concentrations (80 µg ml-1).

  13. Protein–nanoparticle interaction in bioconjugated silver nanoparticles: A transmission electron microscopy and surface enhanced Raman spectroscopy study

    Energy Technology Data Exchange (ETDEWEB)

    Reymond-Laruinaz, Sébastien; Saviot, Lucien; Potin, Valérie; Marco de Lucas, María del Carmen, E-mail: delucas@u-bourgogne.fr

    2016-12-15

    Highlights: • Synthesis of protein-conjugated Ag nanoparticles (NPs) in absence of citrates. • NPs size and protein layer thickness determined by TEM. • SERS spectra showed the chemisorption of proteins on the surface of Ag-NPs. - Abstract: Understanding the mechanisms of interaction between proteins and noble metal nanoparticles (NPs) is crucial to extend the use of NPs in biological applications and nanomedicine. We report the synthesis of Ag-NPs:protein bioconjugates synthesized in total absence of citrates or other stabilizing agents in order to study the NP-protein interaction. Four common proteins (lysozyme, bovine serum albumin, cytochrome-C and hemoglobin) were used in this work. Transmission electron microscopy (TEM) and surface enhanced Raman spectroscopy (SERS) were mainly used to study these bioconjugated NPs. TEM images showed Ag NPs with sizes in the 5–40 nm range. The presence of a protein layer surrounding the Ag NPs was also observed by TEM. Moreover, the composition at different points of single bioconjugated NPs was probed by electron energy loss spectroscopy (EELS). The thickness of the protein layer varies in the 3–15 nm range and the Ag NPs are a few nanometers away. This allowed to obtain an enhancement of the Raman signal of the proteins in the analysis of water suspensions of bioconjugates. SERS results showed a broadening of the Raman bands of the proteins which we attribute to the contribution of different configurations of the proteins adsorbed on the Ag NPs surface. Moreover, the assignment of an intense and sharp peak in the low-frequency range to Ag–N vibrations points to the chemisorption of the proteins on the Ag-NPs surface.

  14. Sintering of oxide-supported Pt and Pd nanoparticles in air studied by in situ TEM

    DEFF Research Database (Denmark)

    Simonsen, Søren Bredmose

    of the sintering mechanisms of nanoparticles is important for making improvements to their long term catalytic activity. Diesel oxidation catalysts are usually composed of noble metal nanoparticles on a complex three-dimensional high surface area oxide. The complex support structure makes it difficult to directly...... observe dynamical processes such as particle sintering with the present state of the art microscope techniques, and consequently it is difficult to relate experimental observations and theoretical sintering models. To reduce the complexity, the present study uses planar model catalysts. These are composed...

  15. Composition of α- F e nanoparticles precipitated from CuFe alloy studied by hyperfine interactions

    Science.gov (United States)

    Kubániová, Denisa; Cesnek, Martin; Milkovi c, Ondrej; Kohout, Jaroslav; Miglierini, Marcel

    2016-12-01

    Iron-based nanoparticles prepared by precipitation from solid solution of saturated binary Cu-Fe alloy were studied by transmission electron microscopy, high-energy X-ray diffraction and Mössbauer spectroscopy. The results showed that the investigated as-prepared nanoparticles contained two phases. The major phase was determined as α- F e and the minor phase as γ- F e 2 O 3. Furthermore, additionally annealed samples in Ar protective atmosphere were investigated. Results showed clear decrease in contribution of α- F e phase and also revealed the presence of various iron oxides (maghemite, magnetite, hematite and wűstite).

  16. Induction of osteogenic differentiation of stem cells via a lyophilized microRNA reverse transfection formulation on a tissue culture plate

    DEFF Research Database (Denmark)

    Wu, Kaimin; Xu, Jie; Liu, Mingzhe

    2013-01-01

    MicroRNA (miRNA) regulation is a novel approach to manipulating the fate of mesenchymal stem cells, but an easy, safe, and highly efficient method of transfection is required. In this study, we developed an miRNA reverse transfection formulation by lyophilizing Lipofectamine 2000-miRNA lipoplexes...... on a tissue culture plate. The lipoplexes can be immobilized on a tissue culture plate with an intact pseudospherical structure and lyophilization without any lyoprotectant. In this study, reverse transfection resulted in highly efficient cellular uptake of miRNA and enabled significant manipulation...

  17. Enhanced effect of nuclear localization signal peptide during ultrasound‑targeted microbubble destruction‑mediated gene transfection.

    Science.gov (United States)

    Cao, Sheng; Zhou, Qing; Chen, Jin-Ling; Jiang, Nan; Wang, Yi-Jia; Deng, Qing; Hu, Bo; Guo, Rui-Qiang

    2017-07-01

    Ultrasound‑targeted microbubble destruction (UTMD) can promote the entry of plasmid DNA (pDNA) into the cell cytoplasm, by increasing the permeability of the cell membrane. But the transfection efficiency remains low due to inability of the pDNA to enter the nucleus. Various methods have been explored to improve the UTMD transfection efficiency, but with little success. In cells, the classic nuclear localization signal (cNLS) peptide is an amino acid sequence that signals proteins that are due for nuclear transport. The present study aimed to investigate whether binding of a cNLS peptide to the pDNA may improve the transfection efficiency of UTMD. Four experimental groups were analyzed: Control group (UTMD + pDNA), group with cNLS (UTMD + pDNA + cNLS), group with mutated NLS (mNLS; UTMD + pDNA + mNLS), and group with cNLS and the nuclear import blocker, wheat germ agglutinin (WGA; UTMD + pDNA + cNLS + WGA). The NLS was labeled by fluorescein isothiocyanate, whereas pDNA was labeled with Cy3. Different molar ratios were tested for the NLS and pDNA combination in order to achieve optimal binding of the two molecules. Human umbilical vein endothelial cells were then transfected using the optimum ultrasonic irradiation parameters and NLS/pDNA molar ratio. At 6 h post‑transfection, the rates of Cy3‑labeled pDNA inside the cells and their nuclei were detected by flow cytometry and laser confocal microscopy, and the cellular vs. nuclear uptake of pDNA was calculated. In order to further evaluate the effect of NLS on UTMD‑mediated gene transfection, the transfection efficiency and relative expression levels of mRNA and protein were detected at 48 h post‑transfection. The results demonstrated that the optimal molar ratio of NLS with pDNA was 104:1. The rates of pDNA successful entry into the cell and nucleus were significantly higher in the cNLS group compared with the control group. The transfection efficiency, and relative expression levels of mRNA and protein

  18. Study on core–shell–shell structured nanoparticles with magnetic and luminescent features: Construction, characterization and oxygen-sensing behavior

    International Nuclear Information System (INIS)

    Min, BU; Wenzhong, Ma

    2013-01-01

    In this paper, we construct core–shell–shell structured nanoparticles, where magnetic Fe 3 O 4 nanoparticles are used as the inner core, mesoporous silica functionalized with phosphorescent Ru(II) complex is used as the outer shell, and the middle shell which is composed of amorphous silica is introduced to minimize the negative effect from the inner core on the sensing probes. The obtained magnetic–luminescent composite nanoparticles are characterized by XRD analysis, IR spectrum, electron microscopy, fluorescence microscopy, thermogravimetric analysis and nitrogen adsorption and desorption, confirming the core–shell–shell structure. The magnetic and photophysical properties of the composite nanoparticles are investigated in detail. Data suggest that the nanoparticles show a smaller saturation magnetization value compared with that of Fe 3 O 4 nanoparticles. The composite namoparticles are red-emitting ones, and the emission is sensitive towards oxygen concentration variations with sensitivity of 4.1 and response time of 7 s. -- Highlights: • Core–shell–shell structured nanoparticles have been constructed. • The nanoparticles have been fully characterized and studied. • The nanoparticles own magnetic, luminescent and oxygen-sensing features. • Good sensitivity, short response time and high photostability have been realized

  19. Preparation of ZnO and ZnS nanoparticles and in-vitro study of their antimicrobial effect

    Directory of Open Access Journals (Sweden)

    Mojtaba Falahati

    2017-06-01

    Full Text Available Zinc sulfide (ZnS & zinc oxide (ZnO nanoparticles was evaluated for their antimicrobial activity against four pathogenic strains. ZnS&ZnO nanoparticles were synthesized by simple aqueous chemical reaction in an aqueous solution. The main advantage of these nanoparticles (size of 10-30 nm was that simply could be prepared by using cheap precursors in a cost effective and high throughput manner. Structural, morphological and chemical composition of the prepared nanoparticles were investigated by X-Ray Diffraction (XRD, Scanning Electron Microscopy (SEM and energy dispersion X-ray dispersive fluorescence spectroscopy (EDAX. The antimicrobial effects of the ZnS&ZnOnanoparticls were studied by serial dilution technique and also by well diffusion technique against four pathogenic microorganism strains of Staphyloccusaureus, Escherichia coli, Pseudomonas aeroginosa and Candidiaalbicans. Both nanoparticles of ZnS&ZnO showed antimicrobial activity against both Gram positive Staphyloccusaureus and Gram negative Escherichia coli and Pseudomonas aeroginosa and fungi of Candidiaalbicans. The best antimicrobial efficacy (as MIC of 50 µg/ml was related to effect of ZnO nanoparticles on Staphyloccusaureus and most resistant pathogen was Candidiaaibicans against ZnS nanoparticles with MIC more than 250 µg/ml. Zinc sulfide (ZnS & zinc oxide (ZnO nanoparticles was evaluated for their antimicrobial activity against four pathogenic strains. ZnS&ZnO nanoparticles were synthesized by simple aqueous chemical reaction in an aqueous solution.

  20. Developing a puncture-free in ovo chicken transfection strategy based on bypassing albumen nucleases.

    Science.gov (United States)

    Amini, Hamid-Reza; Pakdel, Abbas; Shahr-Babak, Hossein Moradi; Eghbalsaied, Shahin

    2017-03-15

    Chicken is a dual-purpose animal important from both agricultural and medical aspects. Even though significant improvements have been made in chicken transgenesis technologies, chicken genome manipulation has not been widely used in developmental biology. This study was aimed to evaluate chicken egg white nuclease properties and thereof plausibility of devising an in vivo transfection technology without causing physical damage to the embryo. First, the nuclease activity of egg albumen was assessed. The egg white nucleases were strongly active in degrading DNA and RNA. The egg white DNase activity was comparable to commercially available DNase-I. Nuclease activities were also assessed after heating, proteinase K, or EDTA treatment. Unlike proteinase K, both heating and EDTA were noticeably effective for the nuclease inactivation. Simultaneous application of lipoplex form of DNA (1 μg pDB2: 3 μl Lipofectamine2000) and EDTA showed a synergistic effect in protection against egg white nucleases. Finally, we injected the lipoplexes with or without EDTA close to the embryo at day0, but outside the embryonic epiblast. Implementation of a scrutinized PCR assay indicated that transfection took place only when EDTA was complemented to the lipoplexes. The transfection rate of day4 embryos and the hatched chicks were 54.5 and 30.0%, respectively. EGFP expression was detected in two out of three transgenic chicks. In conclusion, this study provided a detail analysis of chicken egg albumen nuclease properties and suggested the feasibility of developing a puncture-free handmade technology for transfection of the chicken embryo. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. An in vivo transfection system for inducible gene expression and gene silencing in murine hepatocytes.

    Science.gov (United States)

    Hubner, Eric K; Lechler, Christian; Kohnke-Ertel, Birgit; Zmoos, Anne-Flore; Sage, Julien; Schmid, Roland M; Ehmer, Ursula

    2017-01-01

    Hydrodynamic tail vein injection (HTVI) of transposon-based integration vectors is an established system for stably transfecting mouse hepatocytes in vivo that has been successfully employed to study key questions in liver biology and cancer. Refining the vectors for transposon-mediated hepatocyte transfection will further expand the range of applications of this technique in liver research. In the present study, we report an advanced transposon-based system for manipulating gene expression in hepatocytes in vivo. Transposon-based vector constructs were generated to enable the constitutive expression of inducible Cre recombinase (CreER) together with tetracycline-inducible transgene or miR-small hairpin RNA (shRNA) expression (Tet-ON system). Transposon and transposase expression vectors were co-injected into R26R-mTmG reporter mice by HTVI. Cre-mediated gene recombination was induced by tamoxifen, followed by the administration of doxycycline to drive tetracycline-inducible gene or shRNA expression. Expression was visualized by immunofluorescence staining in livers of injected mice. After HTVI, Cre recombination by tamoxifen led to the expression of membrane-bound green fluorescent protein in transfected hepatocytes. Activation of inducible gene or shRNA expression was detected by immunostaining in up to one-third of transfected hepatocytes, with an efficiency dependent on the promoter driving the Tet-ON system. Our vector system combines Cre-lox mediated gene mutation with inducible gene expression or gene knockdown, respectively. It provides the opportunity for rapid and specific modification of hepatocyte gene expression and can be a useful tool for genetic screening approaches and analysis of target genes specifically in genetically engineered mouse models. Copyright © 2016 John Wiley & Sons, Ltd.

  2. [Expression of human Jagged-1 protein on eukaryotic cells and establishment of stable transfectant cell line].

    Science.gov (United States)

    Gan, Zhi-Hua; Chen, Yu; Yan, Hua; Wang, Kan-Kan

    2010-08-01

    Jagged-1 protein is one of the ligands belonging to Notch signaling pathway. Notch signaling pathway is one of the major signaling pathways mediated by contact between cells and plays an important role to regulate the process of proliferation and differentiation of hematopoietic cells in the hematopoietic microenvironment. To study the biological effect after the combination of receptor and ligand in Notch signaling pathway and the mechanism of Notch signaling pathway in bone marrow stromal cells mediated-drug resistance, a NIH-3T3 cell line over-expressing Jagged-1 protein was constructed for further research purposes. A full coding region of Jagged-1 gene was cloned and inserted into eukaryotic expression plasmid to construct pEGFP-IRES2-Jagged-1 eukaryotic expression vector, then transfected into NIH-3T3 cell line, a mammalian cells. As a result Western blot analysis confirmed that the transfectant NIH-3T3 cells highly expressed Jagged-1 protein and flow cytometry analysis confirmed that the NIH-3T3-pEGFP-IRES2-Jagged-1 cell line over-expressed Jagged-1 protein was monoclonal after screened by selective medium and limiting dilution analysis. It is concluded that the pEGFP-IRES2-Jagged-1 eukaryotic expression vector and a stable transfectant monoclonal NIH-3T3 cell line are successfully established. The construction of the stable transfectant monoclonal NIH-3T3 cell line which overexpressed Jagged-1 protein, provides the conditions to further study the mechanism of the bone marrow stromal cell-mediated drug resistance and to discover the new drug targets.

  3. Effects of Microbubble Size on Ultrasound-Mediated Gene Transfection in Auditory Cells

    Directory of Open Access Journals (Sweden)

    Ai-Ho Liao

    2014-01-01

    Full Text Available Gene therapy for sensorineural hearing loss has recently been used to insert genes encoding functional proteins to preserve, protect, or even regenerate hair cells in the inner ear. Our previous study demonstrated a microbubble- (MB-facilitated ultrasound (US technique for delivering therapeutic medication to the inner ear. The present study investigated whether MB-US techniques help to enhance the efficiency of gene transfection by means of cationic liposomes on HEI-OC1 auditory cells and whether MBs of different sizes affect such efficiency. Our results demonstrated that the size of MBs was proportional to the concentration of albumin or dextrose. At a constant US power density, using 0.66, 1.32, and 2.83 μm albumin-shelled MBs increased the transfection rate as compared to the control by 30.6%, 54.1%, and 84.7%, respectively; likewise, using 1.39, 2.12, and 3.47 μm albumin-dextrose-shelled MBs increased the transfection rates by 15.9%, 34.3%, and 82.7%, respectively. The results indicate that MB-US is an effective technique to facilitate gene transfer on auditory cells in vitro. Such size-dependent MB oscillation behavior in the presence of US plays a role in enhancing gene transfer, and by manipulating the concentration of albumin or dextrose, MBs of different sizes can be produced.

  4. The Effect of Linear PEI on Characteristics and Transfection Efficiency of PEI-Based Cationic Nanoliposomes

    Directory of Open Access Journals (Sweden)

    Mohammad Ramezani

    2011-01-01

    Full Text Available Objective(sThe development of efficient and safe carrier system to transfer DNA into cells is essential in non-viral gene therapy. The aim of the present study was to evaluate the effect of linear polyetheneimine (lPEI (2500 Da on the physicochemical and biological properties of lipopolyplexes constructed from liposomes and lPEI. Materials and MethodsDifferent lipopolymers were synthesized from lPEI and acrylate derivatives. Nanocarriers were composed of the lipids (DOPE, DPPE and DOTAP and the synthesized lipopolymers. After characterization of the prepared vectors by determination of size and zeta potential, transfection activity was tested in Neuro2A cells. Ethidium bromide and MTT test were used to evaluate the DNA condensation ability and cytotoxicity of vectors, respectively. Results Vector’s size ranged from 95 to 337 nm and they had positive charge. The differences in DNA binding properties of lipopolyplexes were not significant. Among lipids, DOTAP showed better impact on transfection efficiency. The highest transfection activity was achieved by liposomal formulation consist of DOTAP and lipopolymer composed of lPEI and hexyl acrylate. The lipopolyplexes showed minimum cytotoxicity to the cultured cells in vitro. Conclusion The results of study confirmed that it is possible to improve gene expression using lipopolyplexes.

  5. Palladium nanoparticles anchored on graphene nanosheets: Methanol, ethanol oxidation reactions and their kinetic studies

    KAUST Repository

    Nagaraju, Doddahalli H.

    2014-12-01

    Palladium nanoparticles decorated graphene (Gra/Pd nanocomposite) was synthesized by simultaneous chemical reduction of graphene oxide and palladium salt in a single step. The negatively charged graphene oxide (GO) facilitates uniform distribution of Pd2+ ions onto its surface. The subsequent reduction by hydrazine hydrate provides well dispersed Pd nanoparticles decorated graphene. Different amount of Pd nanoparticles on graphene was synthesized by changing the volume to weight ratio of GO to PdCl2. X-ray diffraction studies showed FCC lattice of Pd with predominant (1 1 1) plane. SEM and TEM studies revealed that thin graphene nanosheets are decorated by Pd nanoparticles. Raman spectroscopic studies revealed the presence of graphene nanosheets. The electro-catalytic activity of Gra/Pd nanocomposites toward methanol and ethanol oxidation in alkaline medium was evaluated by cyclic voltammetric studies. 1:1 Gra/Pd nanocomposite exhibited good electro-catalytic activity and efficient electron transfer. The kinetics of electron transfer was studied using chronoamperometry. Improved electro-catalytic activity of 1:1 Gra/Pd nanocomposite toward alcohol oxidation makes it as a potential anode for the alcohol fuel cells. © 2014 Elsevier Ltd.

  6. Atorvastatin calcium loaded chitosan nanoparticles: in vitro evaluation and in vivo pharmacokinetic studies in rabbits

    Directory of Open Access Journals (Sweden)

    Abdul Baquee Ahmed

    2015-06-01

    Full Text Available In this study, we prepared atorvastatin calcium (AVST loaded chitosan nanoparticles to improve the oral bioavailability of the drug. Nanoparticles were prepared by solvent evaporation technique and evaluated for its particle size, entrapment efficiency, zeta potential, in vitro release and surface morphology by scanning electron microscopy (SEM. In addition, the pharmacokinetics of AVST from the optimized formulation (FT5 was compared with marketed immediate release formulation (Atorva(r in rabbits. Particle size of prepared nanoparticles was ranged between 179.3 ± 7.12 to 256.8 ± 8.24 nm with a low polydispersity index (PI value. Zeta potential study showed that the particles are stable with positive values between 13.03 ± 0.32 to 46.90 ± 0.49 mV. FT-IR studies confirmed the absence of incompatibility of AVST with excipient used in the formulations. In vitro release study showed that the drug release was sustained for 48 h. Results of pharmacokinetics study showed significant changes in the pharmacokinetic parameter (2.2 fold increase in AUC of the optimized formulation as compared to marketed formulation (Atorva(r. Thus, the developed nanoparticles evidenced the improvement of oral bioavailability of AVST in rabbit model.

  7. Development of an immiscible polymer/polymer/nanoparticle system in order to study the location of nanoparticles at polymer/polymer interface by quantitative optical microscopy

    Science.gov (United States)

    Johansen, Luis Henrique B.; Canto, Leonardo B.; Canevarolo, Sebastião V.

    2015-12-01

    In the past ten years, stabilization of the phase morphology of immiscible polymer blends during melt compounding went through a new perspective by the use of inorganic nanoparticles as compatibilizers. Following the ideas of Ramsden and Pickering, the stabilization of the minor phase in immiscible polymer blends could be achieved with solid nanoparticles located at the interface of the phases, lowering the interfacial tension and acting as a physical barrier to droplet coalescence. In this work, the location of the silica nanoparticle in an immiscible polymer blend is studied using quantitative optical microscopy, measuring the total light scattering, i.e. turbidity, created by the use of hydrophilic and hydrophobic silica nanoparticles (hi-silica and hb-silica, respectively) in an immiscible polymer blend. The light scattering at the polymer/polymer interface is minimized choosing a PS/PC immiscible blend which has minimal difference in their refractive indices. On the other hand, the considerable difference in the refractive index of the chosen polymers and nanosilica would highlight the scattering effect of the silica nanoparticles if located at the polymer/polymer interface. The transmitted light intensity from neat PS/PC blends and some PS/PC/hl-silica systems were similar, showing only a small change in the range of the glass transition temperatures of the two polymers, which is an indication that the silica nanoparticles are dispersed inside the two polymer phases. However, the transmitted light intensity is greatly changed in the system PS/PC/hb-silica, containing the hydrophobic silica, which according to the wetting parameter should have the silica nanoparticles located mainly at the polymer/polymer interface.

  8. Small-Angle Neutron Scattering Study of Interplay of Attractive and Repulsive Interactions in Nanoparticle-Polymer System.

    Science.gov (United States)

    Kumar, Sugam; Aswal, Vinod K; Kohlbrecher, Joachim

    2016-02-16

    The phase behavior of nanoparticle (silica)-polymer (polyethylene glycol) system without and with an electrolyte (NaCl) has been studied. It is observed that nanoparticle-polymer system behaves very differently in the presence of electrolyte. In the absence of electrolyte, the nanoparticle-polymer system remains in one-phase even at very high polymer concentrations. On the other hand, a re-entrant phase behavior is found in the presence of electrolyte, where one-phase (individual) system undergoes two-phase (nanoparticle aggregation) and then back to one-phase with increasing polymer concentration. The regime of two-phase system has been tuned by varying the electrolyte concentration. The polymer concentration range over which the two-phase system exists is significantly enhanced with the increase in the electrolyte concentration. These systems have been characterized by small-angle neutron scattering (SANS) experiments of contrast-marching the polymer to the solvent. The data are modeled using a two-Yukawa potential accounting for both attractive and repulsive parts of the interaction between nanoparticles. The phase behavior of nanoparticle-polymer system is explained by interplay of attractive (polymer-induced attractive depletion between nanoparticles) and repulsive (nanoparticle-nanoparticle electrostatic repulsion and polymer-polymer repulsion) interactions present in the system. In the absence of electrolyte, the strong electrostatic repulsion between nanoparticles dominates over the polymer-induced depletion attraction and the nanoparticle system remains in one-phase. With addition of electrolyte, depletion attraction overcomes electrostatic repulsion at some polymer concentration, resulting into nanoparticle aggregation and two-phase system. Further addition of polymer increases the polymer-polymer repulsion which eventually reduces the strength of depletion and hence re-entrant phase behavior. The effects of varying electrolyte concentration on the phase

  9. Studies on interfacial interactions of TiO2 nanoparticles with ...

    Indian Academy of Sciences (India)

    Administrator

    Studies on interfacial interactions of TiO2 nanoparticles with bacterial cells under light and dark conditions by Swayamprava Dalai et al (pp 371–381). Graphical abstract. Sequence of events occurring in course of cell-NP interaction: Surface adsorption, internalization and bioaccumulation of NPs which leads to stress upon ...

  10. Evaluation of twin-head electrospray nanoparticle disperser for nanotoxicity study

    Science.gov (United States)

    Liu, Qiaoling; Budiman, Thomas; Chen, Da-Ren

    2014-08-01

    With the rapid development of nanotechnology, nanoparticles with various sizes and compositions have been synthesized and proposed for industrial applications. At the same time, the health effects and environmental impacts of nanoparticles become an emerging concern to be addressed. Both in vitro and in vivo studies are of importance to better understand the toxicity of nanoparticles. It is thus essential to have a nanoparticle disperser capable of dispersing individual nanoparticles for these studies. A twin-head electrospray (THES) nanoparticle disperser for animal inhalation exposure studies has recently become commercially available from TSE Systems Inc. Different from the cone-jet electrospray method used in the majority of literature, this particular disperser operates at the multi-jet mode. In this study, we reported our finding on the performance evaluation of the THES disperser with respect to its mass throughput and quality of size distribution of aerosol produced. Three different nanomaterials (TiO2, ZnO, and NiO) were used in this study. It is found that the maximal mass throughput of the studied disperser was achieved by keeping the distance between two opposite spray capillary tips at 3.0 cm, operating the primary carrier-to-capillary sheath flow rates at the ratio of 4:3, and feeding spray suspensions at a flow rate of 20 µl/min. Under the above settings and operations, the highest mass concentration for nano-ZnO was measured at 14.56 mg/m3. Nanoparticle streams with higher concentrations can be further produced by lowering the total carrier gas flow rate and spraying suspensions of higher nanomaterial concentrations. Our study also found that the particle mass throughput of the studied disperser had a good linear relationship with the mass concentration of spray suspension. In addition, the spatial uniformity of nano aerosol distribution in a TSE head-nose-only exposure chamber was investigated. An acceptable nano aerosol uniformity result was

  11. [Transfection Efficiency of Ad5F11p-GFP on CIK and NK-92 Cells and Its Influence on Biological Characteristics].

    Science.gov (United States)

    Xu, Zan-Mei; Lu, Ying; Zhao, Lan-Jun; Liu, Jin; Hu, Xian-Wen; Wu, Chu-Tse; Duan, Hai-Feng

    2016-06-01

    To study transfection efficiency of Ad5F11p-GFP and its influence on biological characteristics of CIK and NK-92 cells in order to predict the application of Ad5F11p vector in immunotherapy. Two kinds of immune cells, cytokine-induced killer (CIK) cells and natural-killer (NK) cell line NK-92 cells, were transfected by Ad5F11p-GFP at different multiplicity of transfection (MOI), and untransfected immune cells were used as negative control. GFP expression was determined by flow cytometry, the cell morphology was observed with microscope, the cell proliferation was analyzed by trypan blue staining, specific cytotoxicity of NK-92 cells was determined by LDH assay. About 90% of transfection efficiency for NK-92 cells could be achieved at a MOI of 25, while the transfection efficiency for CIK was less than 40% at a MOI of 200. In addition, the transfection efficiency basically unchanged at the same MOI for 48 h and 96 h, and the immune cells transfected with the virus trended to form agglomeration, displaying slower proliferation, increase of IFN-γ release and enhancement of tumor killing activity. Ad5F11p- modified NK-92 shows a good prospect for adoptive immunotherapy.

  12. Transfer studies of polystyrene nanoparticles in the ex vivo human placenta perfusion model: key sources of artifacts

    Science.gov (United States)

    Grafmueller, Stefanie; Manser, Pius; Diener, Liliane; Maurizi, Lionel; Diener, Pierre-André; Hofmann, Heinrich; Jochum, Wolfram; Krug, Harald F.; Buerki-Thurnherr, Tina; von Mandach, Ursula; Wick, Peter

    2015-08-01

    Nanotechnology is a rapidly expanding and highly promising new technology with many different fields of application. Consequently, the investigation of engineered nanoparticles in biological systems is steadily increasing. Questions about the safety of such engineered nanoparticles are very important and the most critical subject with regard to the penetration of biological barriers allowing particle distribution throughout the human body. Such translocation studies are technically challenging and many issues have to be considered to obtain meaningful and comparable results. Here we report on the transfer of polystyrene nanoparticles across the human placenta using an ex vivo human placenta perfusion model. We provide an overview of several challenges that can potentially occur in any translocation study in relation to particle size distribution, functionalization and stability of labels. In conclusion, a careful assessment of nanoparticle properties in a physiologically relevant milieu is as challenging and important as the actual study of nanoparticle-cell interactions itself.

  13. Reduced repair of non-dimer photoproducts in a gene transfected into xeroderma pigmentosum cells

    International Nuclear Information System (INIS)

    Protic-Sabljic, Miroslava; Kraemer, K.H.

    1986-01-01

    Cells from patients with the sun sensitive cancer-prone disease, xeroderma pigmentosum (XP) have defective repair of UV damaged DNA with reduced excision of the major photoproduct, the cyclobutane type pyrimidine dimer. Other (non-dimer) photoproducts, have recently been implicated in UV mutagenesis. Utilizing an expression vector host cell reactivation assay, UV damaged transfecting DNA that was treated by in vitro photoreactivation to reverse pyrimidine dimers while not altering other photoproducts was studied. It was found that the reduced expression of a UV damaged transfecting plasmid in XP complementation group A cells is only partially reversed by photoreactivation. E. coli photolyase treatment of pSV2catSVgpt exposed to 100 or 200 J m -2 of 254 nm radiation removed 99% of the T4 endonuclease V sensitive sites. Transfection of XP12BE(SV40) cells with photoreactivated pSV2catSVgpt showed residual inhibition corresponding to 25 to 37% of the lethal hits to the cat gene. This residual inhibition corresponds to the fraction of non-dimer photoproducts induced by UV. This result implies that XP12BE(SV40) cells do not repair most of the non-dimer photoproducts in DNA. (author)

  14. X-ray sensitive strains of CHO cells show decreased frequency of stable transfection

    International Nuclear Information System (INIS)

    Jeggo, P.; Smith, J.

    1987-01-01

    Six X-ray sensitive (xrs) strains of the Chinese hamster ovary cell line have previously been isolated and shown to have a defect in double strand break rejoining. In this study, these strains have been investigated for their ability to take up and integrate foreign DNA. All the xrs strains investigated so far have shown a decreased frequency of stable transfectants compared to their parent line, in experiments using the plasmid pSV2gpt, which contains the selectable bacterial gene, guanine phosphoribosyl transferase. This decreased frequency is observed over a wide range of DNA concentrations (0.1 to 20 μg DNA) but is more pronounced at higher DNA concentrations. In contrast, these xrs strains show the same level of transfection proficiency as the wild type parent using a transient transfection system with a plasmid containing the bacterial CAT (chloramphenicol acetyl transferase) gene. Since the level of CAT activity does not depend on integration of foreign DNA, this suggests that the xrs strains are able to take up the same amount of DNA as the parent strains, but have a defect in the integration of foreign DNA. Since this integration of foreign DNA probably occurs by non-homologous recombination, this may indicate a role of the xrs gene product in this process

  15. Extended gene expression by medium exchange and repeated transient transfection for recombinant protein production enhancement.

    Science.gov (United States)

    Cervera, Laura; Gutiérrez-Granados, Sonia; Berrow, Nicholas Simon; Segura, Maria Mercedes; Gòdia, Francesc

    2015-05-01

    Production of recombinant products in mammalian cell cultures can be achieved by stable gene expression (SGE) or transient gene expression (TGE). The former is based on the integration of a plasmid DNA into the host cell genome allowing continuous gene expression. The latter is based on episomal plasmid DNA expression. Conventional TGE is limited to a short production period of usually about 96 h, therefore limiting productivity. A novel gene expression approach termed extended gene expression (EGE) is explored in this study. The aim of EGE is to prolong the production period by the combination of medium exchange and repeated transfection of cell cultures with plasmid DNA to improve overall protein production. The benefit of this methodology was evaluated for the production of three model recombinant products: intracellular GFP, secreted GFP, and a Gag-GFP virus-like particles (VLPs). Productions were carried out in HEK 293 cell suspension cultures grown in animal-derived component free media using polyethylenimine (PEI) as transfection reagent. Transfections were repeated throughout the production process using different plasmid DNA concentrations, intervals of time, and culture feedin