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Polyethylenimine
Kinds of Polyethylenimine Selected AbstractsSelf-Assembled Multilayers of Polyethylenimine and DNA: Spectrophotometric and Electrochemical Characterization and Application for the Determination of Acridine Orange InteractionELECTROANALYSIS, Issue 15 2009F. Ferreyra Abstract This work deals with the study of the interaction between acridine orange (AO) and calf-thymus double stranded DNA (dsDNA) present in supramolecular architectures built on gold electrodes modified with mercapto-1-propanesulfonic acid (MPS) by self-assembling of polyethylenimine and dsDNA. The optimal conditions for building the supramolecular architecture were obtained from UV-vis spectrophotometric experiments. The electrochemical studies were performed by adsorptive transfer square wave voltammetry from the evaluation of the oxidation signal of AO accumulated within the multistructure. The effect of the number of PEI-dsDNA bilayers (Au/MPS/(PEI-dsDNA)n) on the accumulation and electrooxidation of AO is also discussed. [source] Fabrication of Galactosylated Polyethylenimine and Plasmid DNA Multilayers on poly (D,L -lactic acid) Films for in situ Targeted Gene Transfection,ADVANCED ENGINEERING MATERIALS, Issue 5 2009Yan Hu This study presents surface-mediated targeted in situ gene delivery from gene-tagged poly(D,L -lactic acid) (PDLLA) films, which were fabricated via a layer-by-layer (LbL) assembly technique with galactosylated polyethylenimine (GP) and plasmid DNA (pDNA, pSV-,-galactosidase). A linear growth of GP/pDNA multilayered films was observed. The pDNA was continuously released from multilayered films for over 32,h. The multilayered structure degraded and simultaneously formed GP/pDNA complexes in situ when exposing to a physiological environment. The pDNA was well protected by GP against DNase I digestion within formed GP/pDNA complexes. Our results demonstrated that GP contributes to receptor-mediated targeting for cell uptake and in situ gene transfection. The results reported here are potentially important for gene therapy, surface engineering of biomaterials, tissue engineering and implant technology. [source] Self-Assembled Multilayers of Polyethylenimine and DNA: Spectrophotometric and Electrochemical Characterization and Application for the Determination of Acridine Orange InteractionELECTROANALYSIS, Issue 15 2009F. Ferreyra Abstract This work deals with the study of the interaction between acridine orange (AO) and calf-thymus double stranded DNA (dsDNA) present in supramolecular architectures built on gold electrodes modified with mercapto-1-propanesulfonic acid (MPS) by self-assembling of polyethylenimine and dsDNA. The optimal conditions for building the supramolecular architecture were obtained from UV-vis spectrophotometric experiments. The electrochemical studies were performed by adsorptive transfer square wave voltammetry from the evaluation of the oxidation signal of AO accumulated within the multistructure. The effect of the number of PEI-dsDNA bilayers (Au/MPS/(PEI-dsDNA)n) on the accumulation and electrooxidation of AO is also discussed. [source] On-line biosensors for simultaneous determination of glucose, choline, and glutamate integrated with a microseparation systemELECTROPHORESIS, Issue 18 2003Guoyue Shi Abstract An effective microseparation system integrated with ring-disc electrodes and two microfluidic devices was fabricated for in vivo determination using a microdialysis pump. The major interference of ascorbic acid (AA) was excluded by direct oxidation with ascorbate oxidase. Glucose, glutamate, and choline were successfully determined simultaneously through the biosensors modified with a bilayer of osmium-poly(4-vinylpyridine)gel-horseradish peroxidase (Os-gel-HRP)/glucose oxidase (GOD), glutamate oxidase (GlutaOD) or choline oxidase (ChOD). To stabilize the biosensors, 0.2% polyethylenimine (PEI) was mixed with the oxidases. The cathodic currents of glucose, glutamate, and choline biosensors started to increase after the standard solutions were injected into the microseparation system. The on-line biosensors show a wide calibration range (10,7,10,5 mol/L) with a detection limit of 10,8 mol/L at the working potential of ,50 mV. The variations of glucose, glutamate, and choline were determined simultaneously in a free moving rat when we perfused the medial frontal cortex with 100 ,mol/L N -methyl- D -aspartate (NMDA) solution, which is the agonist of the NMDA receptor. [source] Construction of Polyethyleneimine-,-cyclodextrin/pDNA Multilayer Structure for Improved In Situ Gene Transfection,ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Yan Hu This study reports in situ gene delivery from gene-functionalized poly(D,L -lactic acid) (PDLLA, Mw of around 2.0,×,105,g,mol,1) films, which were constructed via layer-by-layer (LbL) assembly technique with low molecular weight polyethylenimine-,-cyclodextrin (PEI-CD) conjugate and plasmid DNA (pDNA). PEI-CD was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR), respectively. The buildup of multilayered PEI-CD/pDNA pairs onto PDLLA films was monitored with contact angle measurements and UV,Vis spectrometer, respectively. A sustained release of pDNA from multilayered films was observed for 28,h. The mechanism of in situ gene delivery on PDLLA film was investigated in this study as well. Spherical PEI-CD/pDNA complexes were formed and released following the deconstruction of multilayered films, which was confirmed by transmission electron microscopy (TEM) and gel electrophoresis, respectively. Surface mediated in situ gene transfection was achieved when culturing hepatoma G2 (HepG2) and human embryonic kidney 293 (HEK293) onto PEI-CD/pDNA multilayered films. Furthermore, PEI-CD improved the gene transfection efficiency when compared with that of PEI. Such gene-functionalized biomaterial reported here has potential application in tissue engineering and implant technology. [source] Fabrication of Galactosylated Polyethylenimine and Plasmid DNA Multilayers on poly (D,L -lactic acid) Films for in situ Targeted Gene Transfection,ADVANCED ENGINEERING MATERIALS, Issue 5 2009Yan Hu This study presents surface-mediated targeted in situ gene delivery from gene-tagged poly(D,L -lactic acid) (PDLLA) films, which were fabricated via a layer-by-layer (LbL) assembly technique with galactosylated polyethylenimine (GP) and plasmid DNA (pDNA, pSV-,-galactosidase). A linear growth of GP/pDNA multilayered films was observed. The pDNA was continuously released from multilayered films for over 32,h. The multilayered structure degraded and simultaneously formed GP/pDNA complexes in situ when exposing to a physiological environment. The pDNA was well protected by GP against DNase I digestion within formed GP/pDNA complexes. Our results demonstrated that GP contributes to receptor-mediated targeting for cell uptake and in situ gene transfection. The results reported here are potentially important for gene therapy, surface engineering of biomaterials, tissue engineering and implant technology. [source] Multilayer Hybrid Films Consisting of Alternating Graphene and Titania Nanosheets with Ultrafast Electron Transfer and Photoconversion PropertiesADVANCED FUNCTIONAL MATERIALS, Issue 22 2009Kiran Kumar Manga Abstract Alternating graphene (G) and titania (Ti0.91O2) multilayered nanosheets are fabricated using layer-by-layer electrostatic deposition followed by UV irradiation. Successful assemblies of graphene oxide (GO) and titania nanosheets in sequence with polyethylenimine as a linker is confirmed by UV,vis absorption and X-ray diffraction. Photocatalytic reduction of GO into G can be achieved upon UV irradiation. Ultrafast photocatalytic electron transfer between the titania and graphene is demonstrated using femtosecond transient absorption spectroscopy. Efficient exciton dissociation at the interfaces coupled with cross-surface charge percolation allows efficient photocurrent conversion in the multilayered Ti0.91O2/G films. [source] Achieving efficient delivery of morpholino oligos in cultured cellsGENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 3 2001Paul A. Morcos Abstract Summary: One of the many features that make morpholino oligos unique among the antisense structural types is an uncharged backbone. While this feature eliminates the nonspecific interactions of traditional S-oligos, it also renders the morpholino undeliverable via the traditional lipid-based delivery systems. This article describes a highly efficient method of delivering morpholino oligos into adherent and nonadherent cultured cells. In this system, a nonionic morpholino oligo is paired to a complementary DNA "carrier." The DNA is then bound electrostatically to a partially ionized, weakly-basic ethoxylated polyethylenimine (EPEI). This morpholino/DNA/EPEI complex is efficiently endocytosed, and when the pH drops within the endosome, the EPEI more fully ionizes, resulting in permeabilization of the endosomal membrane and release of the morpholino into the cytosol. This article describes optimization of delivery in HeLa cells and provides the basis for delivery in any cultured endocytic cell type. genesis 30:94,102, 2001. © 2001 Wiley-Liss, Inc. [source] PEI,PEG,Chitosan-Copolymer-Coated Iron Oxide Nanoparticles for Safe Gene Delivery: Synthesis, Complexation, and TransfectionADVANCED FUNCTIONAL MATERIALS, Issue 14 2009Forrest M. Kievit Abstract Gene therapy offers the potential of mediating disease through modification of specific cellular functions of target cells. However, effective transport of nucleic acids to target cells with minimal side effects remains a challenge despite the use of unique viral and non-viral delivery approaches. Here, a non-viral nanoparticle gene carrier that demonstrates effective gene delivery and transfection both in vitro and in vivo is presented. The nanoparticle system (NP,CP,PEI) is made of a superparamagnetic iron oxide nanoparticle (NP), which enables magnetic resonance imaging, coated with a novel copolymer (CP,PEI) comprised of short chain polyethylenimine (PEI) and poly(ethylene glycol) (PEG) grafted to the natural polysaccharide, chitosan (CP), which allows efficient loading and protection of the nucleic acids. The function of each component material in this nanoparticle system is illustrated by comparative studies of three nanoparticle systems of different surface chemistries, through material property characterization, DNA loading and transfection analyses, and toxicity assessment. Significantly, NP,CP,PEI demonstrates an innocuous toxic profile and a high level of expression of the delivered plasmid DNA in a C6 xenograft mouse model, making it a potential candidate for safe in vivo delivery of DNA for gene therapy. [source] Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference,HEPATOLOGY, Issue 6 2007Shirish Paranjpe Hepatocyte growth factor (HGF) and its receptor c-Met are involved in liver regeneration. The role of HGF and c-Met in liver regeneration in rat following two-thirds partial hepatectomy (PHx) was investigated using RNA interference to silence HGF and c-Met in separate experiments. A mixture of 2 c-Met-specific short hairpin RNA (ShRNA) sequences, ShM1 and ShM2, and 3 HGF-specific ShRNA, ShH1, ShH3, and ShH4, were complexed with linear polyethylenimine. Rats were injected with the ShRNA/PEI complex 24 hours before and at the time of PHx. A mismatch and a scrambled ShRNA served as negative controls. ShRNA treatment resulted in suppression of c-Met and HGF mRNA and protein compared with that in controls. The regenerative response was assessed by PCNA, mitotic index, and BrdU labeling. Treatment with the ShHGF mixture resulted in moderate suppression of hepatocyte proliferation. Immunohistochemical analysis revealed severe suppression of incorporation of BrdU and complete absence of mitosis in rats treated with ShMet 24 hours after PHx compared with that in controls. Gene array analyses indicated abnormal expression patterns in many cell-cycle- and apoptosis-related genes. The active form of caspase 3 was seen to increase in ShMet-treated rats. The TUNEL assay indicated a slight increase in apoptosis in ShMet-treated rats compared with that in controls. Conclusion: The data indicated that in vivo silencing of c-Met and HGF mRNA by RNA interference in normal rats results in suppression of mRNA and protein, which had a measurable effect on proliferation kinetics associated with liver regeneration. (HEPATOLOGY 2007.) [source] Multilayer Nanocomplexes of Polymer and DNA Exhibit Enhanced Gene Delivery,ADVANCED MATERIALS, Issue 1 2008M. Saul Polymeric-DNA complexes (polyplexes) are constructed with multiple layers of counter-polyions as DNA/polyethylenimine/poly(acrylic acid)/polyethylenimine. The increased association of polyethylenimine achieved by the multilayer approach leads to substantial increases in expression of transgene for reporter plasmids without the need for excess free polymer typically required for non-viral gene delivery. This method of polyplex preparation provides the opportunity to improve transgene expression for gene therapy approaches to disease treatment. [source] Combinatorial Modification of Degradable Polymers Enables Transfection of Human Cells Comparable to Adenovirus,ADVANCED MATERIALS, Issue 19 2007J. Green End-modified poly(,-amino ester)s, easy-to-synthesize degradable polymers, are able to deliver DNA to primary human cells at levels comparable to adenovirus and two orders of magnitude better than the commonly used non-viral vector, polyethylenimine. Small structural changes are found to affect multiple steps of gene delivery including the DNA binding affinity, nanoparticle size, intracellular DNA uptake, and final protein expression. In vivo, these polymer modifications enhance DNA delivery to ovarian tumors. [source] Comparison of the effects of polyethylenimine and maleated polypropylene coupling agents on the properties of cellulose-reinforced polypropylene compositesJOURNAL OF APPLIED POLYMER SCIENCE, Issue 5 2008C. González-Sánchez Abstract The desire to improve the properties of cellulose-reinforced composites while producing them by methods as similar as possible to those used on an industrial scale is one of the driving forces in this field of research. In this work, extensive research for determining the mechanical, thermal, rheological, and physical properties of novel cellulose-reinforced polypropylene composites containing a polyethylenimine (PEI) coupling agent was conducted. A comparison of their properties with those of reference composites without any coupling agent or containing a maleated polypropylene (MAPP) coupling agent was also carried out. The presence of the PEI coupling agent mainly gave rise to a substantial increase in the tensile and flexural strengths and elongations as well as the impact strength, heat deflection temperature (HDT), melt volume flow index, and water absorption of PEI-containing composites in comparison with composites without any coupling agent added. However, the increases achieved in the tensile and flexural composite strengths and HDT were lower than those achieved with the MAPP coupling agent mainly for composites containing 50 wt % cellulose fibers. On the other hand, PEI-containing composites exhibited, in most cases, larger elongations and energies required to break in tensile tests as well as larger impact strengths, melt volume flow indices, and water absorption percentages than MAPP-containing composites. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] In vitro release of complexed pDNA from biodegradable polymer filmsJOURNAL OF APPLIED POLYMER SCIENCE, Issue 1 2008Y. Ramgopal Abstract The controlled delivery of low-molecular weight drugs and proteins from biodegradable polymers has received considerable attention. However, controlled release studies of pDNA from such polymers have not been reported to date. In this study, a plasmid DNA was complexed with the cationic polymer called polyethylenimine (PEI). This gene vector has been shown to be very effective in transfecting cells. The complexed DNA were then incorporated into different types of poly-lactic- co -glycolic acid (PLGA) film; PLGA 53/47 (Mw 90 kDa), 50/50 (Mw 11 kDa, end group is lauryl ester) and 75/25 (Mw 120 kDa). Their release profiles from a buffer solution were studied. An initial (small) burst release of PEI-DNA from film was observed in PLGA 53/47 and 50/50, followed by a plateau phase and finally a rapid erosion-controlled release. For PLGA 50/50, the rapid release started after 14 days; erosion-controlled release for PLGA 53/47 started after 9 days; for PLGA 75/25, the release rate was governed by an initial burst release (10%) followed by a slow release controlled by diffusion. No obvious erosion-controlled release rate was observed for this polymer up to 27 days. Thus, the controlled release of complexed DNA follows the general features exhibited by lower- Mw drugs. This is of significance in designing gene vector matrices that offer the promise of more lasting gene therapy compared with particulate formulations. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] Synthesis and characterization of dexamethasone-conjugated linear polyethylenimine as a gene carrierJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 3 2010Hyunjung Kim Abstract Linear polyethylenimine (25,kDa, LPEI25k) has been shown to be an effective non-viral gene carrier with higher transfection and lower toxicity than branched polyethylenimine (BPEI) of comparable molecular weight. In this study, dexamethasone was conjugated to LPEI25k to improve the efficiency of gene delivery. Dexamethasone is a synthetic glucocorticoid receptor ligand. Dexamethasone-conjugated LPEI25k (LPEI,Dexa) was evaluated as a gene carrier in various cells. Gel retardation assays showed that LPEI,Dexa completely retarded plasmid DNA (pDNA) at a 0.75:1 weight ratio (LPEI/pDNA). LPEI,Dexa had the highest transfection efficiency at a 2:1 weight ratio (LPEI,Dexa/DNA). At this ratio, the size of the LPEI,Dexa/pDNA complex was approximately 125,nm and the zeta potential was 35,mV. LPEI,Dexa had higher transfection efficiency than LPEI and Lipofectamine 2000. In addition, the cytotoxicity of LPEI,Dexa was much lower than that of BPEI (25,kDa, BPEI25k). In conclusion, LPEI,Dexa has a high transfection efficiency and low toxicity and can therefore be used for non-viral gene delivery. J. Cell. Biochem. 110: 743,751, 2010. © 2010 Wiley-Liss, Inc. [source] Synthesis and evaluation of a water-soluble polymer to reduce Ac-225 daughter migrationJOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 2 2007Jonathan Fitzsimmons Abstract The actinium decay chain has been promoted as an in vivo alpha generator for therapy, but migration of daughters from the primary conjugate has lead to increased toxicity away from the target organ. To reduce daughter migration, polyethylenimine (PEI) was used with a primary chelator and secondary chelators. The primary chelator, DOTA, was used to coordinate 225Actinium and secondary chelators-acetate and DTPA, were added to the polymer for coordination of daughters formed by decay. The 225Actinium polymer derivatives containing secondary chelators were found to retain radioactive daughters better than the 225Actinium bond to the primary alone. The retention of 213Bismuth and 209Thallium had the following order from highest retained to lowest DOTA-PEI-DTPA,DOTA-PEI-CH2OO- > DOTA-PEI. The data suggests this polymer approach could be used to reduce daughter migration and has potential for development of actinium labeled radiopharmaceuticals. Copyright © 2007 John Wiley & Sons, Ltd. [source] Phosphothioated oligodeoxynucleotides induce nonspecific effects on neuronal cell adhesion in a growth substrate-dependent manner,JOURNAL OF NEUROSCIENCE RESEARCH, Issue 8 2009Eitan Okun Abstract Synthetic phosphothioated (PTO) oligodeoxynucleotide (ODN) sequences are commonly used for a variety of applications that benefit from nuclease protection. The PTO modification is implemented mainly in antisense ODN, but also in ODN that were shown to activate members of the toll-like receptor (TLR) family such as TLR3 (poly-I:C), TLR8 (ssRNA), and TLR9 (CpG). Neurons are routinely plated on surfaces coated with either cationic substances such as poly-L-ornithine (PLO), polyethylenimine (PEI), poly-L-lysine or ECM components such as laminin, collagen, or fibronectin. We found that PTO-ODN aimed at activating TLR9 induces a non-TLR9-specific detachment phenotype in cortical neurons plated on either laminin or PEI, but not on PLO. This phenotype was correlated with decreased viability and was partially inhibited when caspase-3 was inhibited with Ac-DEVD-CMK. This finding suggests that the use of PTO-ODN can cause nonspecific effects on cell adhesion that could compromise interpretation of data from experiments using PTO-ODN. © 2009 Wiley-Liss, Inc. [source] Colonic delivery of ,-lactamases does not affect amoxicillin pharmacokinetics in ratsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 5 2008Sandrine Bourgeois Abstract Pectin beads containing ,-lactamases were designed for the hydrolysis of colonic residual antibiotics responsible for the emergence of resistance. Beads were prepared by ionotropic gelation in CaCl2 and stabilized by coating with polyethylenimine (PEI) to resist disintegration in the upper GI tract. Particle characterization showed that dried beads had a diameter around 1 mm independently of the presence of PEI. Seven to ten percent (w/w) of PEI was located on bead surface forming a coating layer as observed by scanning electron microscopy. PEI improved considerably bead stability in simulated intestinal medium while affecting slightly the encapsulation efficiency of active ,-lactamases. Coated beads were able to preserve ,-lactamases from premature leakage in the upper GIT whereas, in simulated colonic medium, pectinases induced matrix degradation and reduction of ,-lactamase content especially in beads coated in a 0.8% PEI solution. Finally, the pharmacokinetics of amoxicillin in rat after oral administration was not modified by the co-administration of beads containing ,-lactamases. In conclusion, PEI-coated beads are stable in the upper GIT but remain sensitive to the action of pectinolytic enzymes allowing release of ,-lactamases in a colonic medium without modification of the absorption of a ,-lactam antibiotic when co-administered with loaded beads. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97: 1853,1863, 2008 [source] Maintenance of nonviral vector particle size during the freezing step of the lyophilization process is insufficient for preservation of activity: Insight from other structural indicatorsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 10 2001Marion d.C. Molina Abstract The instability of nonviral vectors as liquid formulations has stimulated considerable interest in developing dehydrated formulations that would be resistant to shipping stresses and could be stored at room temperature. Recently, we reported that high sucrose/DNA ratios are capable of maintaining particle size during the freezing step of the lyophilization process and we suggested that the separation of individual particles within sugar matrices is responsible for the reported protection of nonviral vectors during the freezing step of a typical lyophilization protocol. The purpose of this study was to extend these observations to other nonviral vectors that incorporate different cationic components. Cationic lipid-based complexes composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), with helper lipid cholesterol (Chol) or dioleoylphosphatidyl-ethanolamine (DOPE), showed similar protection by sucrose. Formulations of a polyethylenimine (PEI)-based vector required much higher excipient/DNA ratios for size protection compared with protamine- and lipid-based vectors. At low sucrose/DNA ratios, zeta potentials for all complexes were significantly lowered during freezing. Similar results were obtained at high sucrose/DNA ratios, except for DOTAP,DOPE-containing vectors which maintained zeta potential values comparable to unfrozen controls. The changes in zeta potential values indicate that complexes are altered during freezing despite the maintenance of particle size as determined by light scattering. Furthermore, these changes might explain the observed reduction in transfection activity and provide new information about the effects of physicochemical changes of nonviral vectors during the freezing step of lyophilization. © 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:1445,1455, 2001 [source] Unimolecular micelle derived from hyperbranched polyethylenimine with well-defined hybrid shell of poly(ethylene oxide) and polystyrene: A versatile nanocapsuleJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 3 2010Yong Liang Abstract The synthesis and properties of a macromolecular nanocapsule derived from hyperbranched polyethylenimine (HPEI) with well-defined hybrid shell of poly(ethylene oxide) monomethyl ether (mPEO) and polystyrene (PS) are described. HPEI is treated in sequence with 4-glycidol-2,2,6,6-tetrametyl-piperidin-1-oxyl, succinic anhydride, mPEO, leading to a HPEI derivative compatible with nitroxide-mediated living radical polymerization of styrene, thus a macromolecular nanocapsule, HPEI@PEO/PS, is available with a well-defined and tunable hybrid shell of PEO and PS. Within certain PEO/PS ratio, the nanocapsule is soluble in a number of organic solvents as well as in water. The nanocapsule exists as three layer onion-like particle (HPEI@PS@PEO) in water, whereas in chloroform it exists as a hybrid shell particle (HPEI@PEO/PS), and the particles generally exist in the form of unimolecular micelle. In a biphasic water/chloroform mixture, the nanocapsule can transfer anionic, water-soluble guest from an aqueous phase to the chloroform phase; while when dissolved in water, the nanocapsule can efficiently capture both ionic and apolar solutes. Release of the guest can occur under the stimulus of pH or the switch of medium. This is the first example of a unimolecular micelle that can simultaneously deliver both polar and apolar guests. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 681,691, 2010 [source] Corrosion of ZrB2 Powder During Wet Processing , Analysis and ControlJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 5 2008Sea-Hoon Lee Corrosion behavior of ZrB2 powder during wet processing in water or ethyl alcohol was studied both with and without an organic additive. Incorporation of oxygen and pH change did not intensively occur during static aging of aqueous slurries, but corrosion was enhanced when stirring the slurries. The oxygen content of the powder increased rather rapidly with milling time in ethyl alcohol. The molecular weight of polyethylenimine effected the pH change and oxygen content of ZrB2 powder, after corrosion in water for 18 months. X-ray photoelectron spectroscopy analysis informed that the surface of both the pristine and corroded powders was mainly covered with ZrOH, but a certain amount of Zr,B bonding remained at the powder surface after the wet processing. [source] Antimicrobial photodynamic therapy combined with conventional endodontic treatment to eliminate root canal biofilm infectionLASERS IN SURGERY AND MEDICINE, Issue 1 2007Aguinaldo S. Garcez DDS Abstract Background and Objective To compare the effectiveness of antimicrobial photodynamic therapy (PDT), standard endodontic treatment and the combined treatment to eliminate bacterial biofilms present in infected root canals. Study Design/Materials and Methods Ten single-rooted freshly extracted human teeth were inoculated with stable bioluminescent Gram-negative bacteria, Proteus mirabilis and Pseudomonas aeruginosa to form 3-day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify bacterial burdens. PDT employed a conjugate between polyethylenimine and chlorin(e6) as the photosensitizer (PS) and 660-nm diode laser light delivered into the root canal via a 200-µ fiber, and this was compared and combined with standard endodontic treatment using mechanical debridement and antiseptic irrigation. Results Endodontic therapy alone reduced bacterial bioluminescence by 90% while PDT alone reduced bioluminescence by 95%. The combination reduced bioluminescence by >98%, and importantly the bacterial regrowth observed 24 hours after treatment was much less for the combination (P<0.0005) than for either single treatment. Conclusions Bioluminescence imaging is an efficient way to monitor endodontic therapy. Antimicrobial PDT may have a role to play in optimized endodontic therapy. Lasers Surg. Med. © 2006 Wiley-Liss, Inc. [source] RNA-containing adenovirus/polyethylenimine transfer complexes effectively transduce dendritic cells and induce antigen-specific T cell responsesTHE JOURNAL OF GENE MEDICINE, Issue 4 2004Tatjana C. Gust Abstract Background Dendritic cells (DCs) are the most potent antigen-presenting cells in initiating primary immune responses. Given the unique properties of DCs, gene-modified DCs represent a particularly attractive approach for immunotherapy of diseases such as cancer. Methods Gene-modified DCs were obtained by a receptor-mediated gene delivery system using adenovirus (Ad) particles as ligand and RNA or DNA condensed by polyethylenimine (PEI). In vitro transcribed polyadenylated or non-polyadenylated RNA was used. RNA-transduced DCs were generated expressing chicken ovalbumin (OVA) or chimeric constructs thereof, and compared with DNA-transduced DCs. Results Ad/PEI transfection complexes efficiently delivered RNA into DCs. Such RNA-transduced DCs induced OVA-specific T cell responses more effectively than DNA-transduced DCs. Furthermore, DCs transduced with polyadenylated RNA were more potent in stimulating CD4+ and CD8+ T cell responses than DCs transduced with non-polyadenylated RNA and this was particularly important for CD4+ T cell responses. Conclusions Ad/PEI/RNA transfection is an efficient means for generating RNA-transduced DCs and for stimulating antigen-specific T cell responses. Polyadenylation of RNA enhances CD8+ T cell responses and is essential for CD4+ T cell responses. Copyright © 2004 John Wiley & Sons, Ltd. [source] Lactosylated polyethylenimine for gene transfer into airway epithelial cells: role of the sugar moiety in cell delivery and intracellular trafficking of the complexesTHE JOURNAL OF GENE MEDICINE, Issue 3 2004Stéphanie Grosse Abstract Background As we have previously shown that lactosylated polyethylenimine (PEI) is the most efficient glycosylated PEI for gene transfer into human airway epithelial cells in primary culture, we have studied here the role of the lactose residue in the enhancement of gene transfer efficiency observed with lactosylated PEI as compared with unsubstituted PEI in immortalized (,CFTE29o- cells) and primary human airway epithelial cells. Methods and results After three transfections of 1 h performed daily, 60% of ,CFTE29o- cells were transfected with lactosylated PEI, whereas 25% of cells were transfected with unsubstituted PEI (p < 0.05). Cell viability was 1.8-fold greater with lactosylated PEI as compared with unsubstituted PEI (p < 0.05). As assessed by flow cytometry, the cellular uptake of lactosylated complexes was greater than that of complexes made with unsubstituted PEI (p < 0.05) and involved mostly a receptor-mediated endocytosis. The study of the intracellular trafficking in airway epithelial cells of complexes showed an endosomal and lysosomal accumulation of lactosylated complexes. In the presence of a proton pump inhibitor, the level of lactosylated and unsubstituted PEI-mediated gene expression was reduced more than 20-fold, whereas the cell viability increased to almost 100%. For both complexes, a nuclear localization was observed for less than 5% of intracellular complexes. Conclusions Our results show that the greater gene transfer efficiency observed for lactosylated complexes may be attributed to a higher amount of lactosylated complexes incorporated by airway epithelial cells and a lower cytotoxicity that might be related to reduced endosomolytic properties. However, the lactose residues substituting the PEI did not promote the entry of the plasmid into the nucleus. Copyright © 2004 John Wiley & Sons, Ltd. [source] Metastable and stable states of xanthan polyelectrolyte complexes studied by atomic force microscopyBIOPOLYMERS, Issue 3 2004Gjertrud Maurstad Abstract The compaction of the semiflexible polysaccharide xanthan with selected multi- and polyvalent cations was studied. Polyelectrolyte complexes prepared at concentrations of 1,2 ,g/ml were observed by tapping mode atomic force microscopy. High-molecular-weight xanthan compacted with chitosan yields a blend of mainly toroidal and metastable structures and a small fraction of rod-like species. Polyelectrolyte complexes of xanthan with polyethylenimine and trivalent chromium yielded similar structures or alternatively less well packed species. Racquet-type morphologies were identified as kinetically trapped states occurring on the folding path toward the energetically stable state of the toroids. Thermal annealing yielded a shift of the distribution of xanthan,chitosan morphologies toward this stable state. Ensembles of toroidal and rod-like morphologies of the xanthan,chitosan structures, collected using an asphericity index, were analyzed. The mean height of the toroids increased upon heating, with a selective increase in the height range above 2 nm. It is suggested that the observed metastable structures are formed from the high-molecular-weight fraction of xanthan and that these are driven toward the toroidal state, being a low-energy state, following annealing. Considered a model system for condensation of semiflexible polymers, the compaction of xanthan by chitosan captures the system at various stages in the folding toward a low-energy state and thus allows experimental analyses of these intermediates and their evolution. © 2004 Wiley Periodicals, Inc. Biopolymers, 2004 [source] Calcium phosphate transfection generates mammalian recombinant cell lines with higher specific productivity than polyfectionBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2008Sebastien Chenuet Abstract Transfection with polyethylenimine (PEI) was evaluated as a method for the generation of recombinant Chinese hamster ovary (CHO DG44) cell lines by direct comparison with calcium phosphate-DNA coprecipitation (CaPO4) using both green fluorescent protein (GFP) and a monoclonal antibody as reporter proteins. Following transfection with a GFP expression vector, the proportion of GFP-positive cells as determined by flow cytometry was fourfold higher for the PEI transfection as compared to the CaPO4 transfection. However, the mean level of transient GFP expression for the cells with the highest level of fluorescence was twofold greater for the CaPO4 transfection. Fluorescence in situ hybridization on metaphase chromosomes from pools of cells grown under selective pressure demonstrated that plasmid integration always occurred at a single site regardless of the transfection method. Importantly, the copy number of integrated plasmids was measurably higher in cells transfected with CaPO4. The efficiency of recombinant cell line recovery under selective pressure was fivefold higher following PEI transfection, but the average specific productivity of a recombinant antibody was about twofold higher for the CaPO4-derived cell lines. Nevertheless, no difference between the two transfection methods was observed in terms of the stability of protein production. These results demonstrated the feasibility of generating recombinant CHO-derived cell lines by PEI transfection. However, this method appeared inferior to CaPO4 transfection with regard to the specific productivity of the recovered cell lines. Biotechnol. Bioeng. © 2008 Wiley Periodicals, Inc. [source] Scalable production of adeno-associated virus type 2 vectors via suspension transfection,BIOTECHNOLOGY & BIOENGINEERING, Issue 3 2006Joon Young Park Abstract Vectors derived from adeno-associated virus type 2 (AAV2) are promising gene delivery vehicles, but it is still challenging to get the large number of recombinant adeno-associated virus (rAAV) particles required for large animal and clinical studies. Current transfection technology requires adherent cultures of HEK 293 cells that can only be expanded by preparing multiple culture plates. A single large-scale suspension culture could replace these multiple culture preparations, but there is currently no effective co-transfection scheme for generating rAAV from cells in suspension culture. Here, we weaned HEK 293 cells to suspension culture using hydrogel-coated six-well culture plates and established an efficient transfection strategy suitable for these cells. Then the cultures were gradually scaled up. We used linear polyethylenimine (PEI) to mediate transfection and obtained high transfection efficiencies ranging from 54% to 99%, thereby allowing efficient generation of rAAV vectors. Up to 1013 rAAV particles and, more importantly, up to 1011 infectious particles were generated from a 2-L bioreactor culture. The suspension-transfection strategy of this study facilitates the homogeneous preparation of rAAV at a large scale, and holds further potential as the basis for establishing a manufacturing process in a larger bioreactor. © 2006 Wiley Periodicals, Inc. [source] Microparticle-mediated gene delivery for the enhanced expression of a 19-kDa fragment of merozoite surface protein 1 of Plasmodium falciparumBIOTECHNOLOGY PROGRESS, Issue 1 2010Shan Liu Abstract The 19 kDa carboxyl-terminal fragment of merozoite surface protein 1 (MSP119) is a major component of the invasion-inhibitory response in individual immunity to malaria. A novel ultrasonic atomization approach for the formulation of biodegradable poly(lactic- co -glycolic acid) (PLGA) microparticles of malaria DNA vaccines encoding MSP119 is presented here. After condensing the plasmid DNA (pDNA) molecules with a cationic polymer polyethylenimine (PEI), a 40 kHz ultrasonic atomization frequency was used to formulate PLGA microparticles at a flow rate of 18 mL h,1. High levels of gene expression and moderate cytotoxicity in COS-7 cells were achieved with the condensed pDNA at a nitrogen to phosphate (N/P) ratio of 20, thus demonstrating enhanced cellular uptake and expression of the transgene. The ability of the microparticles to convey pDNA was examined by characterizing the formulated microparticles. The microparticles displayed Z-average hydrodynamic diameters of 1.50,2.10 ,m and zeta potentials of 17.8,23.2 mV. The encapsulation efficiencies were between 78 and 83%, and 76 and 85% of the embedded malaria pDNA molecules were released under physiological conditions in vitro. These results indicate that PLGA-mediated microparticles can be employed as potential gene delivery systems to antigen-presenting cells in the prevention of malaria. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Polyethylenimine-coated albumin nanoparticles for BMP-2 deliveryBIOTECHNOLOGY PROGRESS, Issue 4 2008Sufeng Zhang Abstract Nanoparticle (NP)-based delivery has gained importance for improving the potency of therapeutic agents. The bovine serum albumin (BSA) NPs, obtained by a coacervation process, was modified by electrostatic adsorption of cationic polyethylenimine (PEI) to NP surfaces for delivery of bone-inducing growth factor, bone morphogenetic protein-2 (BMP-2). Different concentrations of PEI were utilized for coating BSA NPs to stabilize the colloidal system and to control the release of BMP-2. The NPs were characterized by size and zeta potential measurements, as well as by Scanning Electron Microscopy and Atomic Force Microscopy. The encapsulation efficiency was typically >90% in all NP preparations. In vitro release kinetics showed that the PEI concentration used for coating the NPs efficiently controlled the release of BMP-2, demonstrating a gradual slowing, sustained release pattern during a 10-day study period. The bioactivity of the encapsulated BMP-2 and the toxicity of the NPs were examined by the alkaline phosphatase (ALP) induction assay and the MTT assay, respectively, using C2C12 cells. The results indicated that PEI was the primary determinant of NP toxicities, and BSA NPs coated with 0.1 mg/mL PEI demonstrated tolerable toxicity, retained the bioactivity of BMP-2, and efficiently slowed the release rate of BMP-2. We conclude that BMP-2 encapsulated in BSA NPs might be an efficient way to deliver the protein for in vivo bone induction. [source] One-Step, Painting-Like Coating Procedures To Make Surfaces Highly and Permanently BactericidalBIOTECHNOLOGY PROGRESS, Issue 2 2006Daewon Park Previously we found that covalent attachment of long-chained, moderately hydrophobic polycations to surfaces of solid objects renders the latter permanently bactericidal. Herein we replaced such surface-specific, multistep immobilization techniques with a single-step, general procedure akin to common painting. Glass or polyethylene slides were briefly dipped into organic solutions of certain optimally hydrophobic N -alkyl-PEI (where PEI stands for branched 750-kDa polyethylenimine) polycations, followed by solvent evaporation. The resultant polycation-coated slides were able to kill on contact all of the encountered bacterial cells, whether the Gram-positive human pathogen Staphylococcus aureus or its Gram-negative brethren Escherichia coli. This biocidal effect was found not to be caused by N -alkyl-PEI molecules leached from the surface. Further examination of the mechanism of this bactericidal action suggested that the surface-deposited N -alkyl-PEI kills bacteria by rupturing their cellular membranes. This conclusion was further supported by studies in which the molecular weight of PEI and the hydrophobicity of the alkyl moiety were varied. [source] | |||||||||||||||||||