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Sample records for electroporation based delivery

  1. Electroporation-based DNA delivery technology

    DEFF Research Database (Denmark)

    Gothelf, A; Gehl, Julie

    2014-01-01

    DNA delivery to for example skin and muscle can easily be performed with electroporation. The method is efficient, feasible, and inexpensive and the future possibilities are numerous. Here we present our protocol for gene transfection to mouse skin using naked plasmid DNA and electric pulses....

  2. Enhanced Delivery of DNA or RNA Vaccines by Electroporation.

    Science.gov (United States)

    Broderick, Kate E; Humeau, Laurent M

    2017-01-01

    Nucleic acid vaccines are a next-generation branch of vaccines which offer major benefits over their conventional protein, bacteria, or viral-based counterparts. However, to be effective in large mammals and humans, an enhancing delivery technology is required. Electroporation is a physical technique which results in improved delivery of large molecules through the cell membrane. In the case of plasmid DNA and mRNA, electroporation enhances both the uptake and expression of the delivered nucleic acids. The muscle is an attractive tissue for nucleic acid vaccination in a clinical setting due to the accessibility and abundance of the target tissue. Historical clinical studies of electroporation in the muscle have demonstrated the procedure to be generally well tolerated in patients. Previous studies have determined that optimized electroporation parameters (such as electrical field intensity, pulse length, pulse width and drug product formulation) majorly impact the efficiency of nucleic acid delivery. We provide an overview of DNA/RNA vaccination in the muscle of mice. Our results suggest that the technique is safe and effective and is highly applicable to a research setting as well as scalable to larger animals and humans.

  3. Micro-/nanofluidics based cell electroporation.

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    Wang, Shengnian; Lee, L James

    2013-01-01

    Non-viral gene delivery has been extensively explored as the replacement for viral systems. Among various non-viral approaches, electroporation has gained increasing attention because of its easy operation and no restrictions on probe or cell type. Several effective systems are now available on the market with reasonably good gene delivery performance. To facilitate broader biological and medical applications, micro-/nanofluidics based technologies were introduced in cell electroporation during the past two decades and their advances are summarized in this perspective. Compared to the commercially available bulk electroporation systems, they offer several advantages, namely, (1) sufficiently high pulse strength generated by a very low potential difference, (2) conveniently concentrating, trapping, and regulating the position and concentration of cells and probes, (3) real-time monitoring the intracellular trafficking at single cell level, and (4) flexibility on cells to be transfected (from single cell to large scale cell population). Some of the micro-devices focus on cell lysis or fusion as well as the analysis of cellular properties or intracellular contents, while others are designed for gene transfection. The uptake of small molecules (e.g., dyes), DNA plasmids, interfering RNAs, and nanoparticles has been broadly examined on different types of mammalian cells, yeast, and bacteria. A great deal of progress has been made with a variety of new micro-/nanofluidic designs to address challenges such as electrochemical reactions including water electrolysis, gas bubble formation, waste of expensive reagents, poor cell viability, low transfection efficacy, higher throughput, and control of transfection dosage and uniformity. Future research needs required to advance micro-/nanofluidics based cell electroporation for broad life science and medical applications are discussed.

  4. Painless skin electroporation as a novel way for insulin delivery.

    Science.gov (United States)

    Wong, Tak-Wah; Chen, Tai-Yu; Huang, Chien-Chun; Tsai, Jui-Chen; Hui, Sek Wen

    2011-09-01

    Rigorous research efforts have been undertaken worldwide to develop a needle-free insulin delivery for many decades with limited success. This translational study aims to deliver insulin through skin with painless electroporation. A recently designed microelectrode array was used to deliver insulin in mice with diabetes under electroporation conditions that are painless and harmless on human skin. Under such condition, a therapeutic amount of insulin was delivered successfully through mouse skin. Electroporation alone increased insulin transport around 100-fold compared with passive diffusion. Increased skin temperature to 40°C for 20 min augmented insulin transport to 237-fold more than the control value. Repeated electroporation showed no harm on human skin. The results indicate the potential of painless delivery of insulin through human skin in future clinical practice.

  5. Microscale Symmetrical Electroporator Array as a Versatile Molecular Delivery System

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    Ouyang, Mengxing; Hill, Winfield; Lee, Jung Hyun; Hur, Soojung Claire

    2017-03-01

    Successful developments of new therapeutic strategies often rely on the ability to deliver exogenous molecules into cytosol. We have developed a versatile on-chip vortex-assisted electroporation system, engineered to conduct sequential intracellular delivery of multiple molecules into various cell types at low voltage in a dosage-controlled manner. Micro-patterned planar electrodes permit substantial reduction in operational voltages and seamless integration with an existing microfluidic technology. Equipped with real-time process visualization functionality, the system enables on-chip optimization of electroporation parameters for cells with varying properties. Moreover, the system’s dosage control and multi-molecular delivery capabilities facilitate intracellular delivery of various molecules as a single agent or in combination and its utility in biological research has been demonstrated by conducting RNA interference assays. We envision the system to be a powerful tool, aiding a wide range of applications, requiring single-cell level co-administrations of multiple molecules with controlled dosages.

  6. In vivo electroporation mediated gene delivery to the beating heart.

    Directory of Open Access Journals (Sweden)

    Erick L Ayuni

    Full Text Available Gene therapy may represent a promising alternative strategy for cardiac muscle regeneration. In vivo electroporation, a physical method of gene transfer, has recently evolved as an efficient method for gene transfer. In the current study, we investigated the efficiency and safety of a protocol involving in vivo electroporation for gene transfer to the beating heart. Adult male rats were anesthetised and the heart exposed through a left thoracotomy. Naked plasmid DNA was injected retrograde into the transiently occluded coronary sinus before the electric pulses were applied. Animals were sacrificed at specific time points and gene expression was detected. Results were compared to the group of animals where no electric pulses were applied. No post-procedure arrhythmia was observed. Left ventricular function was temporarily altered only in the group were high pulses were applied; CK-MB (Creatine kinase and TNT (Troponin T were also altered only in this group. Histology showed no signs of toxicity. Gene expression was highest at day one. Our results provide evidence that in vivo electroporation with an optimized protocol is a safe and effective tool for nonviral gene delivery to the beating heart. This method may be promising for clinical settings especially for perioperative gene delivery.

  7. Efficiency of cellular delivery of antisense peptide nucleic acid by electroporation depends on charge and electroporation geometry

    DEFF Research Database (Denmark)

    Joergensen, Mette; Agerholm-Larsen, Birgit; Nielsen, Peter E

    2011-01-01

    with a functional luciferase readout, we demonstrate that parameters such as peptide nucleic acid (PNA) charge and the method of electroporation have dramatic influence on the efficiency of productive delivery. In a suspended cell electroporation system (cuvettes), a positively charged PNA (+8) was most efficiently...... transferred, whereas charge neutral PNA was more effective in a microtiter plate electrotransfer system for monolayer cells. Surprisingly, a negatively charged (-23) PNA did not show appreciable activity in either system. Findings from the functional assay were corroborated by pulse parameter variations......, polymerase chain reaction, and confocal microscopy. In conclusion, we have found that the charge of PNA and electroporation system combination greatly influences the transfer efficiency, thereby illustrating the complexity of the electroporation mechanism....

  8. Current density imaging sequence for monitoring current distribution during delivery of electric pulses in irreversible electroporation.

    Science.gov (United States)

    Serša, Igor; Kranjc, Matej; Miklavčič, Damijan

    2015-01-01

    Electroporation is gaining its importance in everyday clinical practice of cancer treatment. For its success it is extremely important that coverage of the target tissue, i.e. treated tumor, with electric field is within the specified range. Therefore, an efficient tool for the electric field monitoring in the tumor during delivery of electroporation pulses is needed. The electric field can be reconstructed by the magnetic resonance electric impedance tomography method from current density distribution data. In this study, the use of current density imaging with MRI for monitoring current density distribution during delivery of irreversible electroporation pulses was demonstrated. Using a modified single-shot RARE sequence, where four 3000 V and 100 μs long pulses were included at the start, current distribution between a pair of electrodes inserted in a liver tissue sample was imaged. Two repetitions of the sequence with phases of refocusing radiofrequency pulses 90° apart were needed to acquire one current density image. For each sample in total 45 current density images were acquired to follow a standard protocol for irreversible electroporation where 90 electric pulses are delivered at 1 Hz. Acquired current density images showed that the current density in the middle of the sample increased from first to last electric pulses by 60%, i.e. from 8 kA/m2 to 13 kA/m2 and that direction of the current path did not change with repeated electric pulses significantly. The presented single-shot RARE-based current density imaging sequence was used successfully to image current distribution during delivery of short high-voltage electric pulses. The method has a potential to enable monitoring of tumor coverage by electric field during irreversible electroporation tissue ablation.

  9. Electroporation-based technologies for medicine: principles, applications, and challenges.

    Science.gov (United States)

    Yarmush, Martin L; Golberg, Alexander; Serša, Gregor; Kotnik, Tadej; Miklavčič, Damijan

    2014-07-11

    When high-amplitude, short-duration pulsed electric fields are applied to cells and tissues, the permeability of the cell membranes and tissue is increased. This increase in permeability is currently explained by the temporary appearance of aqueous pores within the cell membrane, a phenomenon termed electroporation. During the past four decades, advances in fundamental and experimental electroporation research have allowed for the translation of electroporation-based technologies to the clinic. In this review, we describe the theory and current applications of electroporation in medicine and then discuss current challenges in electroporation research and barriers to a more extensive spread of these clinical applications.

  10. A high throughput droplet based electroporation system

    Science.gov (United States)

    Yoo, Byeongsun; Ahn, Myungmo; Im, Dojin; Kang, Inseok

    2014-11-01

    Delivery of exogenous genetic materials across the cell membrane is a powerful and popular research tool for bioengineering. Among conventional non-viral DNA delivery methods, electroporation (EP) is one of the most widely used technologies and is a standard lab procedure in molecular biology. We developed a novel digital microfluidic electroporation system which has higher efficiency of transgene expression and better cell viability than that of conventional EP techniques. We present the successful performance of digital EP system for transformation of various cell lines by investigating effects of the EP conditions such as electric pulse voltage, number, and duration on the cell viability and transfection efficiency in comparison with a conventional bulk EP system. Through the numerical analysis, we have also calculated the electric field distribution around the cells precisely to verify the effect of the electric field on the high efficiency of the digital EP system. Furthermore, the parallelization of the EP processes has been developed to increase the transformation productivity. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (Grant Number: 2013R1A1A2011956).

  11. Microfluidic electroporation for delivery of small molecules and genes into cells using a common DC power supply.

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    Wang, Hsiang-Yu; Lu, Chang

    2008-06-15

    Electroporation is an efficient method of introducing foreign impermeant molecules such as drugs and genes into cells. Conventional electroporation has been based on the application of short electrical pulses (electropulsation). Electropulsation requires specialized equipment and cannot be integrated easily with techniques such as electrophoresis which is based on constant voltage. Here we demonstrate the delivery of small molecules and genes into cells, using a microfluidic electroporation technique based on constant direct current (DC) voltage that we developed earlier. We demonstrate the delivery of two molecules into Chinese hamster ovary (CHO-K1) cells: a membrane impermeable nucleic acid dye (SYTOX Green) and a plasmid vector carrying the gene for green fluorescent protein (pEGFP-C1). Our devices can exert field variations to flowing cells that are analogous to the application of single or multiple pulses by having different geometries. We investigate the effects of the electrical parameters and different geometries of the device on the transfection efficiency and cell viability. Our technique provides a simple solution to electroporation-based drug and gene delivery by eliminating the need for a pulse generator. We envision that these simple microscale electroporation devices will have the potential to work in parallel on a microchip platform and such technology will allow high-throughput functional screening of drugs and genes. (c) 2008 Wiley Periodicals, Inc.

  12. What you always needed to know about electroporation based DNA vaccines

    DEFF Research Database (Denmark)

    Gothelf, Anita Birgitte; Gehl, Julie

    2012-01-01

    , and it is foreseen that future DNA vaccination may to a large extent be coupled with and dependent upon electroporation based delivery. Understanding the basic science of electroporation and exploiting knowledge obtained on optimization of DNA electrotransfer to muscle and skin, may greatly augment efforts......Vaccinations are increasingly used to fight infectious disease, and DNA vaccines offer considerable advantages, including broader possibilities for vaccination and lack of need for cold storage. It has been amply demonstrated, that electroporation augments uptake of DNA in both skin and muscle...... on vaccine development. The purpose of this review is to give a succinct but comprehensive overview of electroporation as a delivery modality including electrotransfer to skin and muscle. As well, this review will speculate and discuss future uses for this powerful electrotransfer technology....

  13. Treatment planning of electroporation-based medical interventions: electrochemotherapy, gene electrotransfer and irreversible electroporation

    Science.gov (United States)

    Zupanic, Anze; Kos, Bor; Miklavcic, Damijan

    2012-09-01

    In recent years, cancer electrochemotherapy (ECT), gene electrotransfer for gene therapy and DNA vaccination (GET) and tissue ablation with irreversible electroporation (IRE) have all entered clinical practice. We present a method for a personalized treatment planning procedure for ECT, GET and IRE, based on medical image analysis, numerical modelling of electroporation and optimization with the genetic algorithm, and several visualization tools for treatment plan assessment. Each treatment plan provides the attending physician with optimal positions of electrodes in the body and electric pulse parameters for optimal electroporation of the target tissues. For the studied case of a deep-seated tumour, the optimal treatment plans for ECT and IRE require at least two electrodes to be inserted into the target tissue, thus lowering the necessary voltage for electroporation and limiting damage to the surrounding healthy tissue. In GET, it is necessary to place the electrodes outside the target tissue to prevent damage to target cells intended to express the transfected genes. The presented treatment planning procedure is a valuable tool for clinical and experimental use and evaluation of electroporation-based treatments.

  14. Treatment planning of electroporation-based medical interventions: electrochemotherapy, gene electrotransfer and irreversible electroporation.

    Science.gov (United States)

    Zupanic, Anze; Kos, Bor; Miklavcic, Damijan

    2012-09-07

    In recent years, cancer electrochemotherapy (ECT), gene electrotransfer for gene therapy and DNA vaccination (GET) and tissue ablation with irreversible electroporation (IRE) have all entered clinical practice. We present a method for a personalized treatment planning procedure for ECT, GET and IRE, based on medical image analysis, numerical modelling of electroporation and optimization with the genetic algorithm, and several visualization tools for treatment plan assessment. Each treatment plan provides the attending physician with optimal positions of electrodes in the body and electric pulse parameters for optimal electroporation of the target tissues. For the studied case of a deep-seated tumour, the optimal treatment plans for ECT and IRE require at least two electrodes to be inserted into the target tissue, thus lowering the necessary voltage for electroporation and limiting damage to the surrounding healthy tissue. In GET, it is necessary to place the electrodes outside the target tissue to prevent damage to target cells intended to express the transfected genes. The presented treatment planning procedure is a valuable tool for clinical and experimental use and evaluation of electroporation-based treatments.

  15. Effective delivery of DNA into tumor cells and tissues by electroporation of polymer-DNA complex.

    Science.gov (United States)

    Kang, Jeong-Hun; Toita, Riki; Niidome, Takuro; Katayama, Yoshiki

    2008-07-08

    Electroporation is a useful means for non-viral gene delivery. Here, we investigated the use of electroporation to deliver polymer-DNA complexes into living cells using a protein kinase C (PKC)alpha-responsive polymer. The polymer was complexed with a luciferase-encoding DNA and electroporated into B16 melanoma cells. Gene expression from polymer-DNA complexes was 3- to 5-fold higher than from naked DNA. Moreover, after introduction of the polymer-DNA complex into tissues, luciferase levels were >2-fold higher in B16 melanoma tumors than in normal skin tissue. These results suggest that the combination of our polymer and electroporation is useful for the effective delivery of DNA into tumors.

  16. Electroporation for drug and gene delivery in the clinic: doctors go electric

    DEFF Research Database (Denmark)

    Gehl, J.

    2008-01-01

    Electroporation is a unique system for drug and gene delivery, as it is possible to very specifically target certain tissues within the body with whatever drug, gene, isotope, or other product is desired in a specific situation. An increasing number of clinical trials are being launched, and soph......, and sophistication of equipment and protocols continues. This chapter reviews present knowledge from clinical trials, describes important issues in the patient management when using electroporation, and outlines future perspectives of the technology Udgivelsesdato: 2008...

  17. Painless electroporation with a new needle-free microelectrode array to enhance transdermal drug delivery.

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    Wong, Tak-Wah; Chen, Ching-Hung; Huang, Chien-Chun; Lin, Cheng-De; Hui, Sek-Wen

    2006-02-21

    A microelectrode array was designed to minimize the pain sensation of electroporation for enhancing transdermal drug delivery. The influence of the size of the electrode-skin contact area and of the distance between electrodes on the pain sensation was tested on human volunteers. The pain level decreased with the dimension of electrode-skin contact area and with inter-electrode distance. When both reached about 0.5 mm, the pain level was not perceptible even at the threshold of transdermal electroporation level of sixty electric pulses at 150 V, 1 ms at 1-10 Hz. An array of 11 x 11 alternately connected electrodes with 0.6 x 0.6 mm dimension was fabricated. The electric thresholds for effective drug delivery, using toluidine blue O as a marker on mouse skin, was found to be the same for microelectrode arrays as for larger electrodes and wider inter-electrode distances. In vivo transdermal electroporation using microelectrode array with 180 pulses of 150 V, 0.2 ms at 1 Hz, followed by 30 min methotrexate (MTX) occlusion increased more than 4 fold the systemic MTX level in mice. The results demonstrated the potential of painless delivery of significant amounts of chemotherapeutic agents through skin with the new electrode arrays in a clinical setting.

  18. Non-Invasive Delivery of dsRNA into De-Waxed Tick Eggs by Electroporation

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    Ruiz, Newton; de Abreu, Leonardo Araujo; Parizi, Luís Fernando; Kim, Tae Kwon; Mulenga, Albert; Braz, Gloria Regina Cardoso; Vaz, Itabajara da Silva; Logullo, Carlos

    2015-01-01

    RNA interference-mediated gene silencing was shown to be an efficient tool for validation of targets that may become anti-tick vaccine components. Here, we demonstrate the application of this approach in the validation of components of molecular signaling cascades, such as the Protein Kinase B (AKT) / Glycogen Synthase Kinase (GSK) axis during tick embryogenesis. It was shown that heptane and hypochlorite treatment of tick eggs can remove wax, affecting corium integrity and but not embryo development. Evidence of AKT and GSK dsRNA delivery into de-waxed eggs of via electroporation is provided. Primers designed to amplify part of the dsRNA delivered into the electroporated eggs dsRNA confirmed its entry in eggs. In addition, it was shown that electroporation is able to deliver the fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI). To confirm gene silencing, a second set of primers was designed outside the dsRNA sequence of target gene. In this assay, the suppression of AKT and GSK transcripts (approximately 50% reduction in both genes) was demonstrated in 7-day-old eggs. Interestingly, silencing of GSK in 7-day-old eggs caused 25% reduction in hatching. Additionally, the effect of silencing AKT and GSK on embryo energy metabolism was evaluated. As expected, knockdown of AKT, which down regulates GSK, the suppressor of glycogen synthesis, decreased glycogen content in electroporated eggs. These data demonstrate that electroporation of de-waxed R. microplus eggs could be used for gene silencing in tick embryos, and improve the knowledge about arthropod embryogenesis. PMID:26091260

  19. Non-Invasive Delivery of dsRNA into De-Waxed Tick Eggs by Electroporation.

    Directory of Open Access Journals (Sweden)

    Newton Ruiz

    Full Text Available RNA interference-mediated gene silencing was shown to be an efficient tool for validation of targets that may become anti-tick vaccine components. Here, we demonstrate the application of this approach in the validation of components of molecular signaling cascades, such as the Protein Kinase B (AKT/Glycogen Synthase Kinase (GSK axis during tick embryogenesis. It was shown that heptane and hypochlorite treatment of tick eggs can remove wax, affecting corium integrity and but not embryo development. Evidence of AKT and GSK dsRNA delivery into de-waxed eggs of via electroporation is provided. Primers designed to amplify part of the dsRNA delivered into the electroporated eggs dsRNA confirmed its entry in eggs. In addition, it was shown that electroporation is able to deliver the fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI. To confirm gene silencing, a second set of primers was designed outside the dsRNA sequence of target gene. In this assay, the suppression of AKT and GSK transcripts (approximately 50% reduction in both genes was demonstrated in 7-day-old eggs. Interestingly, silencing of GSK in 7-day-old eggs caused 25% reduction in hatching. Additionally, the effect of silencing AKT and GSK on embryo energy metabolism was evaluated. As expected, knockdown of AKT, which down regulates GSK, the suppressor of glycogen synthesis, decreased glycogen content in electroporated eggs. These data demonstrate that electroporation of de-waxed R. microplus eggs could be used for gene silencing in tick embryos, and improve the knowledge about arthropod embryogenesis.

  20. Education on electrical phenomena involved in electroporation-based therapies and treatments: a blended learning approach.

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    Čorović, Selma; Mahnič-Kalamiza, Samo; Miklavčič, Damijan

    2016-04-07

    Electroporation-based applications require multidisciplinary expertise and collaboration of experts with different professional backgrounds in engineering and science. Beginning in 2003, an international scientific workshop and postgraduate course electroporation based technologies and treatments (EBTT) has been organized at the University of Ljubljana to facilitate transfer of knowledge from leading experts to researches, students and newcomers in the field of electroporation. In this paper we present one of the integral parts of EBTT: an e-learning practical work we developed to complement delivery of knowledge via lectures and laboratory work, thus providing a blended learning approach on electrical phenomena involved in electroporation-based therapies and treatments. The learning effect was assessed via a pre- and post e-learning examination test composed of 10 multiple choice questions (i.e. items). The e-learning practical work session and both of the e-learning examination tests were carried out after the live EBTT lectures and other laboratory work. Statistical analysis was performed to compare and evaluate the learning effect measured in two groups of students: (1) electrical engineers and (2) natural scientists (i.e. medical doctors, biologists and chemists) undergoing the e-learning practical work in 2011-2014 academic years. Item analysis was performed to assess the difficulty of each item of the examination test. The results of our study show that the total score on the post examination test significantly improved and the item difficulty in both experimental groups decreased. The natural scientists reached the same level of knowledge (no statistical difference in total post-examination test score) on the post-course test take, as do electrical engineers, although the engineers started with statistically higher total pre-test examination score, as expected. The main objective of this study was to investigate whether the educational content the e

  1. Electroporation-based gene therapy: recent evolution in the mechanism description and technology developments.

    Science.gov (United States)

    Mir, Lluis M

    2014-01-01

    Thirty years after the publication of the first report on gene electrotransfer in cultured cells by the delivery of delivering electric pulses, this technology is starting to be applied to humans. In 2008, at the time of the publication of the first edition of this book, reversible cell electroporation for gene transfer and gene therapy (nucleic acids electrotransfer) was at a cross roads in its development. In 5 years, basic and applied developments have brought gene electrotransfer into a new status. Present knowledge on the effects of cell exposure to appropriate electric field pulses, particularly at the level of the cell membrane, is reported here, as an introduction to the large range of applications described in this book. The importance of the models of electric field distribution in tissues and of the correct choice of electrodes and applied voltages is highlighted, as well as the large range of new specialized electrodes, developed also in the frame of the other electroporation-based treatments (electrochemotherapy). Indeed, electric pulses are now routinely applied for localized drug delivery in the treatment of solid tumors by electrochemotherapy. The mechanisms involved in DNA electrotransfer, which include cell electropermeabilization and DNA electrophoresis, are also surveyed: noticeably, the first molecular description of the crossing of a lipid membrane by a nucleic acid was reported in 2012. The progress in the understanding of cell electroporation as well as developments of technological aspects, in silico, in vitro and in vivo, have contributed to bring gene electrotransfer development to the clinical stage. However, spreading of the technology will require not only more clinical trials but also further homogenization of the protocols and the preparation and validation of Standard Operating Procedures.

  2. Close-field electroporation gene delivery using the cochlear implant electrode array enhances the bionic ear.

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    Pinyon, Jeremy L; Tadros, Sherif F; Froud, Kristina E; Y Wong, Ann C; Tompson, Isabella T; Crawford, Edward N; Ko, Myungseo; Morris, Renée; Klugmann, Matthias; Housley, Gary D

    2014-04-23

    The cochlear implant is the most successful bionic prosthesis and has transformed the lives of people with profound hearing loss. However, the performance of the "bionic ear" is still largely constrained by the neural interface itself. Current spread inherent to broad monopolar stimulation of the spiral ganglion neuron somata obviates the intrinsic tonotopic mapping of the cochlear nerve. We show in the guinea pig that neurotrophin gene therapy integrated into the cochlear implant improves its performance by stimulating spiral ganglion neurite regeneration. We used the cochlear implant electrode array for novel "close-field" electroporation to transduce mesenchymal cells lining the cochlear perilymphatic canals with a naked complementary DNA gene construct driving expression of brain-derived neurotrophic factor (BDNF) and a green fluorescent protein (GFP) reporter. The focusing of electric fields by particular cochlear implant electrode configurations led to surprisingly efficient gene delivery to adjacent mesenchymal cells. The resulting BDNF expression stimulated regeneration of spiral ganglion neurites, which had atrophied 2 weeks after ototoxic treatment, in a bilateral sensorineural deafness model. In this model, delivery of a control GFP-only vector failed to restore neuron structure, with atrophied neurons indistinguishable from unimplanted cochleae. With BDNF therapy, the regenerated spiral ganglion neurites extended close to the cochlear implant electrodes, with localized ectopic branching. This neural remodeling enabled bipolar stimulation via the cochlear implant array, with low stimulus thresholds and expanded dynamic range of the cochlear nerve, determined via electrically evoked auditory brainstem responses. This development may broadly improve neural interfaces and extend molecular medicine applications.

  3. Comparison of small interfering RNA (siRNA) delivery into bovine monocyte-derived macrophages by transfection and electroporation.

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    Jensen, Kirsty; Anderson, Jennifer A; Glass, Elizabeth J

    2014-04-15

    The manipulation of the RNA interference pathway using small interfering RNA (siRNA) has become the most frequently used gene silencing method. However, siRNA delivery into primary cells, especially primary macrophages, is often considered challenging. Here we report the investigation of the suitability of two methodologies: transient transfection and electroporation, to deliver siRNA targeted against the putative immunomodulatory gene Mediterranean fever (MEFV) into primary bovine monocyte-derived macrophages (bMDM). Eleven commercial transfection reagents were investigated with variable results with respect to siRNA uptake, target gene knock-down, cell toxicity and type I interferon (IFN) response induction. Three transfection reagents: Lipofectamine 2000, Lipofectamine RNAiMAX and DharmaFECT 3, were found to consistently give the best results. However, all the transfection reagents tested induced an IFN response in the absence of siRNA, which could be minimized by reducing the transfection reagent incubation period. In addition, optimized siRNA delivery into bMDM by electroporation achieved comparable levels of target gene knock-down as transient transfection, without a detectable IFN response, but with higher levels of cell toxicity. The optimized transient transfection and electroporation methodologies may provide a starting point for optimizing siRNA delivery into macrophages derived from other species or other cells considered difficult to investigate with siRNA. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  4. A modified single-cell electroporation method for molecule delivery into a motile protist, Euglena gracilis.

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    Ohmachi, Masashi; Fujiwara, Yoshie; Muramatsu, Shuki; Yamada, Koji; Iwata, Osamu; Suzuki, Kengo; Wang, Dan Ohtan

    2016-11-01

    Single-cell transfection is a powerful technique for delivering chemicals, drugs, or probes into arbitrary, specific single cells. This technique is especially important when the analysis of molecular function and cellular behavior in individual microscopic organisms such as protists requires the precise identification of the target cell, as fluorescence labeling of bulk populations makes tracking of individual motile protists virtually impossible. Herein, we have modified current single-cell electroporation techniques for delivering fluorescent markers into single Euglena gracilis, a motile photosynthetic microalga. Single-cell electroporation introduced molecules into individual living E. gracilis cells after a negative pressure was applied through a syringe connected to the micropipette to the target cell. The new method achieves high transfection efficiency and viability after electroporation. With the new technique, we successfully introduced a variety of molecules such as GFP, Alexa Fluor 488, and exciton-controlled hybridization-sensitive fluorescent oligonucleotide (ECHO) RNA probes into individual motile E. gracilis cells. We demonstrate imaging of endogenous mRNA in living E. gracilis without interfering with their physiological functions, such as swimming or division, over an extended period of time. Thus the modified single-cell electroporation technique is suitable for delivering versatile functional molecules into individual motile protists. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy – spatial and time dependent distribution

    Directory of Open Access Journals (Sweden)

    Dachs Gabi U

    2004-11-01

    Full Text Available Abstract Background Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation-assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. Methods Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. Results Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. Conclusions Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous

  6. Perspectives on Transdermal Electroporation

    Directory of Open Access Journals (Sweden)

    Kevin Ita

    2016-03-01

    Full Text Available Transdermal drug delivery offers several advantages, including avoidance of erratic absorption, absence of gastric irritation, painlessness, noninvasiveness, as well as improvement in patient compliance. With this mode of drug administration, there is no pre-systemic metabolism and it is possible to increase drug bioavailability and half-life. However, only a few molecules can be delivered across the skin in therapeutic quantities. This is because of the hindrance provided by the stratum corneum. Several techniques have been developed and used over the last few decades for transdermal drug delivery enhancement. These include sonophoresis, iontophoresis, microneedles, and electroporation. Electroporation, which refers to the temporary perturbation of the skin following the application of high voltage electric pulses, has been used to increase transcutaneous flux values by several research groups. In this review, transdermal electroporation is discussed and the use of the technique for percutaneous transport of low and high molecular weight compounds described. This review also examines our current knowledge regarding the mechanisms of electroporation and safety concerns arising from the use of this transdermal drug delivery technique. Safety considerations are especially important because electroporation utilizes high voltage pulses which may have deleterious effects in some cases.

  7. Synergistic Combination of Electrolysis and Electroporation for Tissue Ablation.

    Science.gov (United States)

    Stehling, Michael K; Guenther, Enric; Mikus, Paul; Klein, Nina; Rubinsky, Liel; Rubinsky, Boris

    2016-01-01

    Electrolysis, electrochemotherapy with reversible electroporation, nanosecond pulsed electric fields and irreversible electroporation are valuable non-thermal electricity based tissue ablation technologies. This paper reports results from the first large animal study of a new non-thermal tissue ablation technology that employs "Synergistic electrolysis and electroporation" (SEE). The goal of this pre-clinical study is to expand on earlier studies with small animals and use the pig liver to establish SEE treatment parameters of clinical utility. We examined two SEE methods. One of the methods employs multiple electrochemotherapy-type reversible electroporation magnitude pulses, designed in such a way that the charge delivered during the electroporation pulses generates the electrolytic products. The second SEE method combines the delivery of a small number of electrochemotherapy magnitude electroporation pulses with a low voltage electrolysis generating DC current in three different ways. We show that both methods can produce lesion with dimensions of clinical utility, without the need to inject drugs as in electrochemotherapy, faster than with conventional electrolysis and with lower electric fields than irreversible electroporation and nanosecond pulsed ablation.

  8. Electroporation-mediated in vivo gene delivery of the Na+/K+-ATPase pump reduced lung injury in a mouse model of lung contusion.

    Science.gov (United States)

    Machado-Aranda, David A; Suresh, M V; Yu, Bi; Raghavendran, Krishnan

    2012-01-01

    Lung contusion (LC) is an independent risk factor for acute respiratory distress syndrome. The final common pathway in ARDS involves accumulation of fluid in the alveoli. In this study, we demonstrate the application of a potential gene therapy approach by delivering the Na+/K+-ATPase pump subunits in a murine model of LC. We hypothesized that restoring the activity of the pump will result in removal of excess alveolar fluid and additionally reduce inflammation. Under anesthesia, C57/BL6 mice were struck along the right posterior axillary line 1 cm above the costal margin with a cortical contusion impactor. Immediately afterward, 100 μg of plasmid DNA coding for the α,β of the Na+/K+-ATPase pump were instilled into the lungs (LC-electroporation-pump group). Contusion only (LC-only) and a sham saline instillation group after contusion were used as controls (LC-electroporation-sham). By using a BTX 830 electroporator, eight electrical pulses of 200 V/cm field strength were applied transthoracically. Mice were killed at 24 hours, 48 hours, and 72 hours after delivery. Bronchial alveolar lavage was recollected to measure albumin and cytokines by enzyme-linked immunosorbent assay. Pulmonary compliance was measured, and lungs were subject to histopathologic analysis. After the electroporation and delivery of genes coding for the α,β subunits of the Na+/K+-ATPase pump, there was a significant mitigation of acute lung injury as evidenced by reduction in bronchial alveolar lavage levels of albumin, improved pressure volume curves, and reduced inflammation seen on histology. Electroporation-mediated gene transfer of the subunits of the Na+/K+-ATPase pump enhanced recovery from acute inflammatory lung injury after LC.

  9. Comparative evaluation of transmembrane ion transport due to monopolar and bipolar nanosecond, high-intensity electroporation pulses based on full three-dimensional analyses

    Science.gov (United States)

    Hu, Q.; Joshi, R. P.

    2017-07-01

    Electric pulse driven membrane poration finds applications in the fields of biomedical engineering and drug/gene delivery. Here we focus on nanosecond, high-intensity electroporation and probe the role of pulse shape (e.g., monopolar-vs-bipolar), multiple electrode scenarios, and serial-versus-simultaneous pulsing, based on a three-dimensional time-dependent continuum model in a systematic fashion. Our results indicate that monopolar pulsing always leads to higher and stronger cellular uptake. This prediction is in agreement with experimental reports and observations. It is also demonstrated that multi-pronged electrode configurations influence and increase the degree of cellular uptake.

  10. Ex vivo and in silico feasibility study of monitoring electric field distribution in tissue during electroporation based treatments.

    Directory of Open Access Journals (Sweden)

    Matej Kranjc

    Full Text Available Magnetic resonance electrical impedance tomography (MREIT was recently proposed for determining electric field distribution during electroporation in which cell membrane permeability is temporary increased by application of an external high electric field. The method was already successfully applied for reconstruction of electric field distribution in agar phantoms. Before the next step towards in vivo experiments is taken, monitoring of electric field distribution during electroporation of ex vivo tissue ex vivo and feasibility for its use in electroporation based treatments needed to be evaluated. Sequences of high voltage pulses were applied to chicken liver tissue in order to expose it to electric field which was measured by means of MREIT. MREIT was also evaluated for its use in electroporation based treatments by calculating electric field distribution for two regions, the tumor and the tumor-liver region, in a numerical model based on data obtained from clinical study on electrochemotherapy treatment of deep-seated tumors. Electric field distribution inside tissue was successfully measured ex vivo using MREIT and significant changes of tissue electrical conductivity were observed in the region of the highest electric field. A good agreement was obtained between the electric field distribution obtained by MREIT and the actual electric field distribution in evaluated regions of a numerical model, suggesting that implementation of MREIT could thus enable efficient detection of areas with insufficient electric field coverage during electroporation based treatments, thus assuring the effectiveness of the treatment.

  11. Calcium Electroporation

    DEFF Research Database (Denmark)

    Frandsen, Stine Krog; Gibot, Laure; Madi, Moinecha

    2015-01-01

    BACKGROUND: Calcium electroporation describes the use of high voltage electric pulses to introduce supraphysiological calcium concentrations into cells. This promising method is currently in clinical trial as an anti-cancer treatment. One very important issue is the relation between tumor cell kill...... efficacy-and normal cell sensitivity. METHODS: Using a 3D spheroid cell culture model we have tested the effect of calcium electroporation and electrochemotherapy using bleomycin on three different human cancer cell lines: a colorectal adenocarcinoma (HT29), a bladder transitional cell carcinoma (SW780......), and a breast adenocarcinoma (MDA-MB231), as well as on primary normal human dermal fibroblasts (HDF-n). RESULTS: The results showed a clear reduction in spheroid size in all three cancer cell spheroids three days after treatment with respectively calcium electroporation (p

  12. Modeling of electric field distribution in tissues during electroporation.

    Science.gov (United States)

    Corovic, Selma; Lackovic, Igor; Sustaric, Primoz; Sustar, Tomaz; Rodic, Tomaz; Miklavcic, Damijan

    2013-02-21

    Electroporation based therapies and treatments (e.g. electrochemotherapy, gene electrotransfer for gene therapy and DNA vaccination, tissue ablation with irreversible electroporation and transdermal drug delivery) require a precise prediction of the therapy or treatment outcome by a personalized treatment planning procedure. Numerical modeling of local electric field distribution within electroporated tissues has become an important tool in treatment planning procedure in both clinical and experimental settings. Recent studies have reported that the uncertainties in electrical properties (i.e. electric conductivity of the treated tissues and the rate of increase in electric conductivity due to electroporation) predefined in numerical models have large effect on electroporation based therapy and treatment effectiveness. The aim of our study was to investigate whether the increase in electric conductivity of tissues needs to be taken into account when modeling tissue response to the electroporation pulses and how it affects the local electric distribution within electroporated tissues. We built 3D numerical models for single tissue (one type of tissue, e.g. liver) and composite tissue (several types of tissues, e.g. subcutaneous tumor). Our computer simulations were performed by using three different modeling approaches that are based on finite element method: inverse analysis, nonlinear parametric and sequential analysis. We compared linear (i.e. tissue conductivity is constant) model and non-linear (i.e. tissue conductivity is electric field dependent) model. By calculating goodness of fit measure we compared the results of our numerical simulations to the results of in vivo measurements. The results of our study show that the nonlinear models (i.e. tissue conductivity is electric field dependent: σ(E)) fit experimental data better than linear models (i.e. tissue conductivity is constant). This was found for both single tissue and composite tissue. Our results of

  13. The Effect of Electroporation of a Lyotroic Liquid Crystal Genistein-Based Formulation in the Recovery of Murine Melanoma Lesions

    Directory of Open Access Journals (Sweden)

    Corina Danciu

    2015-07-01

    Full Text Available A lamellar lyotropic liquid crystal genistein-based formulation (LLC-Gen was prepared in order to increase the aqueous solubility of the lipophilic phytocompound genistein. The formulation was applied locally, in a murine model of melanoma, with or without electroporation. The results demonstrated that, when the formulation was applied by electroporation, the tumors appeared later. During the 21 days of the experiment, the LLC-Gen formulation decreased the tumor volume, the amount of melanin and the degree of erythema, but when electroporation was applied, all these parameters indicated a better prognosis even (lower tumor volume, amount of melanin and degree of erythema. Although hematoxylin–eosin (HE staining confirmed the above events, application of the LLC-Gen formulation by electroporation did not lead to a significant effect in terms of the serum concentrations of the protein S100B and serum neuron specific enolase (NSE, or the tissue expression of the platelet-derived growth factor receptor β (PDGFRβ antibody.

  14. Predicting electroporation of cells in an inhomogeneous electric field based on mathematical modeling and experimental CHO-cell permeabilization to propidium iodide determination.

    Science.gov (United States)

    Dermol, Janja; Miklavčič, Damijan

    2014-12-01

    High voltage electric pulses cause electroporation of the cell membrane. Consequently, flow of the molecules across the membrane increases. In our study we investigated possibility to predict the percentage of the electroporated cells in an inhomogeneous electric field on the basis of the experimental results obtained when cells were exposed to a homogeneous electric field. We compared and evaluated different mathematical models previously suggested by other authors for interpolation of the results (symmetric sigmoid, asymmetric sigmoid, hyperbolic tangent and Gompertz curve). We investigated the density of the cells and observed that it has the most significant effect on the electroporation of the cells while all four of the mathematical models yielded similar results. We were able to predict electroporation of cells exposed to an inhomogeneous electric field based on mathematical modeling and using mathematical formulations of electroporation probability obtained experimentally using exposure to the homogeneous field of the same density of cells. Models describing cell electroporation probability can be useful for development and presentation of treatment planning for electrochemotherapy and non-thermal irreversible electroporation. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. siRNA delivery into cultured primary human myoblasts--optimization of electroporation parameters and theoretical analysis.

    Science.gov (United States)

    Lojk, Jasna; Mis, Katarina; Pirkmajer, Sergej; Pavlin, Mojca

    2015-12-01

    Introduction of genetic material into muscle tissue has been extensively researched, including isolation and in vitro expansion of primary myoblasts as a potential source of cells for skeletal and heart muscle tissue engineering applications. In this study, we optimized the electroporation protocol for introduction of short interfering ribonucleic acid (siRNA) against messenger RNA for Hypoxia Inducible Factor 1α (HIF-1α) into cultured primary human myoblasts. We established optimal pulsing protocol for siRNA electro transfection, and theoretically analyzed the effect of electric field and pulse duration on silencing efficiency and electrophoretic displacement of siRNA. Silencing of HIF-1α was determined with quantitative polymerase chain reaction and Western Blot. The most efficient silencing (71% knockdown) was achieved with 8 × 2 ms pulses, E = 0.6 kV/cm. Viability was determined immediately, 1 h and 48 h after electroporation. In general, there was a trade-off between efficient silencing and preserved viability. Electric field and pulse duration are crucial parameters for silencing, since both increase membrane permeabilization and electrophoretic transfer of siRNA. Short-term viability showed immediate toxicity of pulses due to membrane damage, while indirect effects on cell proliferation were observed after 48 h. Presented results are important for faster optimization of electroporation parameters for ex vivo electrotransfer of short RNA molecules into primary human myoblasts. © 2015 Wiley Periodicals, Inc.

  16. Handbook of electroporation

    CERN Document Server

    2017-01-01

    This major reference work is a one-shot knowledge base on electroporation and the use of pulsed electric fields of high intensity and their use in biology, medicine, biotechnology, and food and environmental technologies. The Handbook offers a widespread and well-structured compilation of 156 chapters ranging from the foundations to applications in industry and hospital. It is edited and written by most prominent researchers in the field. With regular updates and growing in its volume it is suitable for academic readers and researchers regardless of their disciplinary expertise, and will also be accessible to students and serious general readers. The Handbook's 276 authors have established scholarly credentials and come from a wide range of disciplines. This is crucially important in a highly interdisciplinary field of electroporation and the use of pulsed electric fields of high intensity and its applications in different fields from medicine, biology, food proce ssing, agriculture, process engineering, en...

  17. Electroporation-induced electrosensitization.

    Directory of Open Access Journals (Sweden)

    Olga N Pakhomova

    Full Text Available BACKGROUND: Electroporation is a method of disrupting the integrity of cell membrane by electric pulses (EPs. Electrical modeling is widely employed to explain and study electroporation, but even most advanced models show limited predictive power. No studies have accounted for the biological consequences of electroporation as a factor that alters the cell's susceptibility to forthcoming EPs. METHODOLOGY/PRINCIPAL FINDINGS: We focused first on the role of EP rate for membrane permeabilization and lethal effects in mammalian cells. The rate was varied from 0.001 to 2,000 Hz while keeping other parameters constant (2 to 3,750 pulses of 60-ns to 9-µs duration, 1.8 to 13.3 kV/cm. The efficiency of all EP treatments was minimal at high rates and started to increase gradually when the rate decreased below a certain value. Although this value ranged widely (0.1-500 Hz, it always corresponded to the overall treatment duration near 10 s. We further found that longer exposures were more efficient irrespective of the EP rate, and that splitting a high-rate EP train in two fractions with 1-5 min delay enhanced the effects severalfold. CONCLUSIONS/SIGNIFICANCE: For varied experimental conditions, EPs triggered a delayed and gradual sensitization to EPs. When a portion of a multi-pulse exposure was delivered to already sensitized cells, the overall effect markedly increased. Because of the sensitization, the lethality in EP-treated cells could be increased from 0 to 90% simply by increasing the exposure duration, or the exposure dose could be reduced twofold without reducing the effect. Many applications of electroporation can benefit from accounting for sensitization, by organizing the exposure either to maximize sensitization (e.g., for sterilization or, for other applications, to completely or partially avoid it. In particular, harmful side effects of electroporation-based therapies (electrochemotherapy, gene therapies, tumor ablation include convulsions

  18. Feasibility of catheter-directed intraluminal irreversible electroporation of porcine ureter and acute outcomes in response to increasing energy delivery.

    Science.gov (United States)

    Srimathveeravalli, Govindarajan; Silk, Mikhail; Wimmer, Thomas; Monette, Sebastien; Kimm, Simon; Maybody, Majid; Solomon, Stephen B; Coleman, Jonathan; Durack, Jeremy C

    2015-07-01

    To evaluate the feasibility of focal intraluminal irreversible electroporation (IRE) in the ureter with a novel electrode catheter and to study the treatment effects in response to increasing pulse strength. Five IRE treatment settings were each evaluated twice for the ablation of normal ureter in 5 Yorkshire pigs (n = 1-4 ablations per animal; total of 10 ablations) with the use of a prototype device under ultrasound and fluoroscopic guidance. Animals received unilateral or bilateral treatment, limited to a maximum of 2 ablations in any 1 ureter. Treatment was delivered with increasing pulse strength (from 1,000 V to 3,000 V in increments of 500 V) while keeping the pulse duration (100 μs) and number of pulses (n = 90) constant. Ureter patency was assessed with antegrade ureteropyelography immediately following treatment. Animals were euthanized within 4 hours after treatment, and treated urinary tract was harvested for histopathologic analysis with hematoxylin and eosin and Masson trichrome stains. IRE was successfully performed in all animals, without evidence of ureteral perforation. Hematoxylin and eosin analysis of IRE treatments demonstrated full-thickness ablation at higher field strengths (mucosa to the adventitia). Masson trichrome stains showed preservation of connective tissue at all field strengths. Intraluminal catheter-directed IRE ablation is feasible and produces full-thickness ablation of normal ureters. There was no evidence of lumen perforation even at the maximum voltages evaluated. Copyright © 2015 SIR. Published by Elsevier Inc. All rights reserved.

  19. An electrically active microneedle array for electroporation

    Science.gov (United States)

    Choi, Seong-O; Kim, Yeu Chun; Park, Jung-Hwan; Hutcheson, Joshua; Gill, Harvinder S.; Yoon, Yong-Kyu; Prausnitz, Mark R.; Allen, Mark G.

    2010-01-01

    We have designed and fabricated a microneedle array with electrical functionality with the final goal of electroporating skin’s epidermal cells to increase their transfection by DNA vaccines. The microneedle array was made of polymethylmethacrylate (PMMA) by micromolding technology from a master PDMS mold, followed by metal deposition, patterning using laser ablation, and electrodeposition. This microneedle array possessed sufficient mechanical strength to penetrate human skin in vivo and was also able to electroporate both red blood cells and human prostate cancer cells as an in vitro model to demonstrate cell membrane permeabilization. A model to predict the effective volume for electroporation with respect to applied voltages was constructed from finite element simulation. This study demonstrates the mechanical and electrical functionalities of the first MEMS-fabricated microneedle array for electroporation, designed for DNA vaccine delivery. PMID:20012696

  20. Segmentation of hepatic vessels from MRI images for planning of electroporation-based treatments in the liver.

    Science.gov (United States)

    Marcan, Marija; Pavliha, Denis; Music, Maja Marolt; Fuckan, Igor; Magjarevic, Ratko; Miklavcic, Damijan

    2014-09-01

    Electroporation-based treatments rely on increasing the permeability of the cell membrane by high voltage electric pulses delivered to tissue via electrodes. To ensure that the whole tumor is covered by the sufficiently high electric field, accurate numerical models are built based on individual patient geometry. For the purpose of reconstruction of hepatic vessels from MRI images we searched for an optimal segmentation method that would meet the following initial criteria: identify major hepatic vessels, be robust and work with minimal user input. We tested the approaches based on vessel enhancement filtering, thresholding, and their combination in local thresholding. The methods were evaluated on a phantom and clinical data. Results show that thresholding based on variance minimization provides less error than the one based on entropy maximization. Best results were achieved by performing local thresholding of the original de-biased image in the regions of interest which were determined through previous vessel-enhancement filtering. In evaluation on clinical cases the proposed method scored in average sensitivity of 93.68%, average symmetric surface distance of 0.89 mm and Hausdorff distance of 4.04 mm. The proposed method to segment hepatic vessels from MRI images based on local thresholding meets all the initial criteria set at the beginning of the study and necessary to be used in treatment planning of electroporation-based treatments: it identifies the major vessels, provides results with consistent accuracy and works completely automatically. Whether the achieved accuracy is acceptable or not for treatment planning models remains to be verified through numerical modeling of effects of the segmentation error on the distribution of the electric field.

  1. Microfluidic Screening of Electric Fields for Electroporation

    Science.gov (United States)

    Garcia, Paulo A.; Ge, Zhifei; Moran, Jeffrey L.; Buie, Cullen R.

    2016-01-01

    Electroporation is commonly used to deliver molecules such as drugs, proteins, and/or DNA into cells, but the mechanism remains poorly understood. In this work a rapid microfluidic assay was developed to determine the critical electric field threshold required for inducing bacterial electroporation. The microfluidic device was designed to have a bilaterally converging channel to amplify the electric field to magnitudes sufficient to induce electroporation. The bacterial cells are introduced into the channel in the presence of SYTOX®, which fluorescently labels cells with compromised membranes. Upon delivery of an electric pulse, the cells fluoresce due to transmembrane influx of SYTOX® after disruption of the cell membranes. We calculate the critical electric field by capturing the location within the channel of the increase in fluorescence intensity after electroporation. Bacterial strains with industrial and therapeutic relevance such as Escherichia coli BL21 (3.65 ± 0.09 kV/cm), Corynebacterium glutamicum (5.20 ± 0.20 kV/cm), and Mycobacterium smegmatis (5.56 ± 0.08 kV/cm) have been successfully characterized. Determining the critical electric field for electroporation facilitates the development of electroporation protocols that minimize Joule heating and maximize cell viability. This assay will ultimately enable the genetic transformation of bacteria and archaea considered intractable and difficult-to-transfect, while facilitating fundamental genetic studies on numerous diverse microbes. PMID:26893024

  2. DNA Vaccine Electroporation and Molecular Adjuvants

    Science.gov (United States)

    2016-03-16

    vaccination, delivery methods, electroporation, molecular adjuvants, intramuscular, intradermal 1. Introduction The Filoviridae family of viruses is...a malaria DNA vaccine by immunization with a needle-free jet device. Vaccine, 2001. 20(1-2): p. 275-80. 20. Choi, A.H., et al., Protection of mice

  3. An Infrared Actin Probe for Deep-Cell Electroporation-Based Single-Molecule Speckle (eSiMS Microscopy

    Directory of Open Access Journals (Sweden)

    Sawako Yamashiro

    2017-07-01

    Full Text Available Single-molecule speckle (SiMS microscopy is a powerful method to directly elucidate biochemical reactions in live cells. However, since the signal from an individual fluorophore is extremely faint, the observation area by epi-fluorescence microscopy is restricted to the thin cell periphery to reduce autofluorescence, or only molecules near the plasma membrane are visualized by total internal reflection fluorescence (TIRF microscopy. Here, we introduce a new actin probe labeled with near infrared (NIR emissive CF680R dye for easy-to-use, electroporation-based SiMS microscopy (eSiMS for deep-cell observation. CF680R-labeled actin (CF680R-actin incorporated into actin structures and showed excellent brightness and photostability suitable for single-molecule imaging. Importantly, the intensity of autofluorescence with respect to SiMS brightness was reduced to approximately 13% compared to DyLight 550-labeled actin (DL550-actin. CF680R-actin enabled the monitoring of actin SiMS in actomyosin bundles associated with adherens junctions (AJs located at 3.5–4 µm above the basal surfaces of epithelial monolayers. These favorable properties of CF680R-actin extend the application of eSiMS to actin turnover and flow analyses in deep cellular structures.

  4. Web-based tool for visualization of electric field distribution in deep-seated body structures and planning of electroporation-based treatments.

    Science.gov (United States)

    Marčan, Marija; Pavliha, Denis; Kos, Bor; Forjanič, Tadeja; Miklavčič, Damijan

    2015-01-01

    Treatments based on electroporation are a new and promising approach to treating tumors, especially non-resectable ones. The success of the treatment is, however, heavily dependent on coverage of the entire tumor volume with a sufficiently high electric field. Ensuring complete coverage in the case of deep-seated tumors is not trivial and can in best way be ensured by patient-specific treatment planning. The basis of the treatment planning process consists of two complex tasks: medical image segmentation, and numerical modeling and optimization. In addition to previously developed segmentation algorithms for several tissues (human liver, hepatic vessels, bone tissue and canine brain) and the algorithms for numerical modeling and optimization of treatment parameters, we developed a web-based tool to facilitate the translation of the algorithms and their application in the clinic. The developed web-based tool automatically builds a 3D model of the target tissue from the medical images uploaded by the user and then uses this 3D model to optimize treatment parameters. The tool enables the user to validate the results of the automatic segmentation and make corrections if necessary before delivering the final treatment plan. Evaluation of the tool was performed by five independent experts from four different institutions. During the evaluation, we gathered data concerning user experience and measured performance times for different components of the tool. Both user reports and performance times show significant reduction in treatment-planning complexity and time-consumption from 1-2 days to a few hours. The presented web-based tool is intended to facilitate the treatment planning process and reduce the time needed for it. It is crucial for facilitating expansion of electroporation-based treatments in the clinic and ensuring reliable treatment for the patients. The additional value of the tool is the possibility of easy upgrade and integration of modules with new

  5. Optimal Electroporation Condition for Small Interfering RNA Transfection into MDA-MB-468 Cell Line

    Directory of Open Access Journals (Sweden)

    Rita Arabsolghar

    2012-09-01

    Full Text Available Background: Electroporation is a valuable tool for small interfering RNA (siRNA delivery into cells because it efficiently transforms a wide variety of cell types. Since electroporation condition for each cell type must be determined experimentally, this study presents an optimal electroporation strategy to reproducibly and efficiently transfect MDA-MB 468 human breast cancer cell with siRNA. Methods: To identify the best condition, the cells were firstly electroporated without siRNA and cell viability was determined by trypan blue and MTT assays. Then siRNA transfection in the best condition was performed. Western blot analysis was used for monitoring successful siRNA transfection. Results: The best condition for electroporation of this cell line was 220 volt and 975 µF in exponential decay using the Gene Pulser X cell electroporation system. Our data demonstrated that by using proper electroporation condition, DNA methyl transferase mRNA was silenced by 10 nmol DNMT1 siRNA in MDA-MB 468 cells when compared with negative control siRNA electroporation. Analysis of cell viability demonstrated that optimal electroporation condition resulted in 74% and 78% cell viability by trypan blue staining and MTT assay, respectively. Conclusion: Transfection of the MDA-MB-468 breast cancer cell line with siRNA in the obtained electroporation condition was successful and resulted in effective gene silencing and high cellular viability.

  6. Albumin-based drug delivery

    DEFF Research Database (Denmark)

    Larsen, Maja Thim; Kuhlmann, Matthias; Hvam, Michael Lykke

    2016-01-01

    binding sites, cellular receptor engagement, and a long circulatory half-life due to interaction with the recycling neonatal Fc receptor. Exploitation of these properties promotes albumin as an attractive candidate for half-life extension and targeted intracellular delivery of drugs attached by covalent...... conjugation, genetic fusions, association or ligand-mediated association. This review will give an overview of albumin-based products with focus on the natural biological properties and molecular interactions that can be harnessed for the design of a next-generation drug delivery platform.......The effectiveness of a drug is dependent on accumulation at the site of action at therapeutic levels, however, challenges such as rapid renal clearance, degradation or non-specific accumulation requires drug delivery enabling technologies. Albumin is a natural transport protein with multiple ligand...

  7. Albumin-based drug delivery

    DEFF Research Database (Denmark)

    Larsen, Maja Thim; Kuhlmann, Matthias; Hvam, Michael Lykke

    2016-01-01

    The effectiveness of a drug is dependent on accumulation at the site of action at therapeutic levels, however, challenges such as rapid renal clearance, degradation or non-specific accumulation requires drug delivery enabling technologies. Albumin is a natural transport protein with multiple ligand...... binding sites, cellular receptor engagement, and a long circulatory half-life due to interaction with the recycling neonatal Fc receptor. Exploitation of these properties promotes albumin as an attractive candidate for half-life extension and targeted intracellular delivery of drugs attached by covalent...... conjugation, genetic fusions, association or ligand-mediated association. This review will give an overview of albumin-based products with focus on the natural biological properties and molecular interactions that can be harnessed for the design of a next-generation drug delivery platform....

  8. Optimized DNA electroporation for primary human T cell engineering.

    Science.gov (United States)

    Zhang, Zhang; Qiu, Shunfang; Zhang, Xiaopeng; Chen, Wei

    2018-01-30

    Effective gene-delivery systems for primary human T cell engineering are useful tools for both basic research and clinical immunotherapy applications. Pseudovirus-based systems and electro-transfection are the most popular strategies for genetic material transduction. Compared with viral-particle-mediated approaches, electro-transfection is theoretically safer, because it does not promote transgene integration into the host genome. Additionally, the simplicity and speed of the procedure increases the attractiveness of electroporation. Here, we developed and optimized an electro-transfection method for the production of engineered chimeric antigen receptor (CAR)-T cells. Stimulation of T cells had the greatest effect on their transfection, with stimulation of cells for up to 3 days substantially improving transfection efficiency. Additionally, the strength of the external electric field, input cell number, and the initial amount of DNA significantly affected transfection performance. The voltage applied during electroporation affected plasmid permeation and was negatively correlated with the number of viable cells after electroporation. Moreover, higher plasmid concentration increased the proportion of positively transfected cells, but decreased cell viability, and for single-activated cells, higher cell density enhanced their viability. We evaluated the effects of two clinically relevant factors, serum supplementation in the culture medium and cryopreservation immediately after the isolation of peripheral blood lymphocytes. Our findings showed that our protocol performed well using xeno-free cultured, fresh T cells, with application resulting in a lower but acceptable transfection efficiency of cells cultured with fetal bovine serum or thawed cells. Furthermore, we described an optimized procedure to generate CAR-T cells within 6 days and that exhibited cytotoxicity toward targeted cells. Our investigation of DNA electro-transfection for the use in human primary

  9. Image-guided Electro-assisted Drug Delivery: Comparison Between Two Types of Electrodes

    Directory of Open Access Journals (Sweden)

    Biliana Nikolova

    2015-06-01

    Full Text Available Electroporation-based cancer treatment techniques are currently after active investigations in the field of drug delivery, optimization of electrical parameters and elucidation of the exact mechanisms at a molecular level. The present study is designed to investigate the exact in vivo redistribution and persistence of nanoparticles in the tumor tissue of colon-cancer grafted mice after electroporation with two different kinds of electrodes. The aim of the study is to avoid artifacts during electroporation due to accumulation of nanoparticles in the surrounding non-cancer tissues. The isolated electrodes are appropriate for the treatment of 3-dimensional tumors and have a large potential in this field.

  10. Recent Trends on Micro/Nanofluidic Single Cell Electroporation

    Directory of Open Access Journals (Sweden)

    Tuhin Subhra Santra

    2013-09-01

    Full Text Available The behaviors of cell to cell or cell to environment with their organelles and their intracellular physical or biochemical effects are still not fully understood. Analyzing millions of cells together cannot provide detailed information, such as cell proliferation, differentiation or different responses to external stimuli and intracellular reaction. Thus, single cell level research is becoming a pioneering research area that unveils the interaction details in high temporal and spatial resolution among cells. To analyze the cellular function, single cell electroporation can be conducted by employing a miniaturized device, whose dimension should be similar to that of a single cell. Micro/nanofluidic devices can fulfill this requirement for single cell electroporation. This device is not only useful for cell lysis, cell to cell fusion or separation, insertion of drug, DNA and antibodies inside single cell, but also it can control biochemical, electrical and mechanical parameters using electroporation technique. This device provides better performance such as high transfection efficiency, high cell viability, lower Joule heating effect, less sample contamination, lower toxicity during electroporation experiment when compared to bulk electroporation process. In addition, single organelles within a cell can be analyzed selectively by reducing the electrode size and gap at nanoscale level. This advanced technique can deliver (in/out biomolecules precisely through a small membrane area (micro to nanoscale area of the single cell, known as localized single cell membrane electroporation (LSCMEP. These articles emphasize the recent progress in micro/nanofluidic single cell electroporation, which is potentially beneficial for high-efficient therapeutic and delivery applications or understanding cell to cell interaction.

  11. Avoiding nerve stimulation in irreversible electroporation: a numerical modeling study

    Science.gov (United States)

    Mercadal, Borja; Arena, Christopher B.; Davalos, Rafael V.; Ivorra, Antoni

    2017-10-01

    Electroporation based treatments consist in applying one or multiple high voltage pulses to the tissues to be treated. As an undesired side effect, these pulses cause electrical stimulation of excitable tissues such as nerves and muscles. This increases the complexity of the treatments and may pose a risk to the patient. To minimize electrical stimulation during electroporation based treatments, it has been proposed to replace the commonly used monopolar pulses by bursts of short bipolar pulses. In the present study, we have numerically analyzed the rationale for such approach. We have compared different pulsing protocols in terms of their electroporation efficacy and their capability of triggering action potentials in nerves. For that, we have developed a modeling framework that combines numerical models of nerve fibers and experimental data on irreversible electroporation. Our results indicate that, by replacing the conventional relatively long monopolar pulses by bursts of short bipolar pulses, it is possible to ablate a large tissue region without triggering action potentials in a nearby nerve. Our models indicate that this is possible because, as the pulse length of these bipolar pulses is reduced, the stimulation thresholds raise faster than the irreversible electroporation thresholds. We propose that this different dependence on the pulse length is due to the fact that transmembrane charging for nerve fibers is much slower than that of cells treated by electroporation because of their geometrical differences.

  12. The influence of soft layer electrokinetics on bacterial electroporation

    Science.gov (United States)

    Moran, Jeffrey; Dingari, Naga Neehar; Buie, Cullen

    2015-11-01

    Electroporation of mammalian cells has received a significant amount of theoretical attention over the last decade because of its ability to deliver biologically active molecules into cells using short and strong electric field pulses. However, application of the same theory to bacterial electroporation presents significant challenges because of the presence of charged soft layers around bacteria. The soft layer charge distribution has been found to significantly influence bacterial electrophoretic mobility and polarizability because it alters the electric potential spatial distribution around the cell envelope. In addition, the RC charging time scale of both the soft layer and electric double layer is of the order of microseconds, which is also of similar order of magnitude as the pore creation time scale. Therefore in this study, we investigate the influence of soft layer electrokinetics on the spatial pore distribution and the temporal pore radius evolution during bacteria electroporation, which are quantitative measures of a bacterium's amenability to electroporation. The study will have significant impact on designing and optimizing bacteria electroporation platforms for gene and drug delivery applications.

  13. Optimizing in vivo gene transfer into mouse corpus cavernosum by use of surface electroporation

    Science.gov (United States)

    Song, Kang-Moon; Choi, Min Ji; Kwon, Mi-Hye; Ghatak, Kalyan; Park, Soo-Hwan; Ryu, Dong-Soo; Ryu, Ji-Kan

    2015-01-01

    Purpose Electroporation is known to enhance the efficiency of gene transfer through a transient increase in cell membrane permeability. The aim of this study was to determine the optimal conditions for in vivo electroporation-mediated gene delivery into mouse corpus cavernosum. Materials and Methods Diabetes was induced in C57BL/6 mice by intraperitoneal injections of streptozotocin. After intracavernous injection of pCMV-Luc (100 µg/40 µL), different electroporation settings (5-50 V, 8-16 pulses with a duration of 40-100 ms) were applied to the penis to establish the optimal conditions for electroporation. Gene expression was evaluated by luciferase assay. We also assessed the undesired consequences of electroporation by visual inspection and hematoxylin-eosin staining of penile tissue. Results Electroporation profoundly induced gene expression in the corpus cavernosum tissue of normal mice in a voltage-dependent manner. We observed electrical burn scars in the penis of normal mice who received electroporation with eight 40-ms pulses at a voltage of 50 V and sixteen 40-ms pulses, eight 100-ms pulses, and sixteen 100-ms pulses at a voltage of 30 V. No detectable burn scars were noted in normal mice stimulated with eight 40-ms pulses at a voltage of 30 V. Electroporation also significantly induced gene expression in diabetic mice stimulated with 40-ms pulse at a voltage of 30 V without injury to the penis. Conclusions We have established the optimal electroporation conditions for maximizing gene transfer into the corpus cavernosum of mice while avoiding damage to the erectile tissue. The electroporation-mediated gene delivery technique will be a valuable tool for gene therapy in the field of erectile dysfunction. PMID:25763123

  14. The Influence of Soft Layer Electrokinetics on Electroporation of Gram-positive Bacteria

    Science.gov (United States)

    Dingari, Naga Neehar; Moran, Jeffrey L.; Garcia, Paulo A.; Buie, Cullen R.

    2016-11-01

    Bacterial electroporation involves subjecting cells to intense ( 10 kV/cm) electric pulses, to open pores on the cell membrane for intracellular delivery of exogenous molecules. Its high efficiency in genetic transformation makes it an attractive tool for synthetic biology. While mammalian cell electroporation has received extensive theoretical and experimental investigation, bacterial electroporation has received markedly less attention. In this work, we develop a theoretical model of electroporation for gram-positive bacteria, taking into account the effect of the bacterial cell envelope on the cell's response to an electroporation pulse. We model the influence of the cell wall charge on the electrokinetic transport (and hence the pore properties) around the bacterial cell envelope using the Poisson-Nernst-Planck equations. Further, we account for the influence of the cell wall's mechanical elasticity on the pore radius evolution during electroporation, which is typically neglected in mammalian cell electroporation. This yields valuable information about favorable conditions for pore formation and will enable designing optimal platforms for bacteria electroporation.

  15. Transformation of methylotrophic bacteria by electroporation.

    Science.gov (United States)

    Gliesche, C G

    1997-02-01

    An efficient system for electroporation of the methylotrophic bacteria Hyphomicrobium facilis, Hyphomicrobium denitrificans, Methylobacillus glycogenes, Methylobacterium extorquens, and Methylophilus methylotrophus is described. It could be demonstrated that vectors based on the broad-host-range plasmid pBBR1 could be transferred into these strains. Plasmid pBBR1KAN (3.9 kb), a kanamycin-resistant derivative of pBBR1, was suitable for transformation experiments in these methylotrophic bacteria. Transformation efficiencies up to 10(4) transformants/microgram plasmid pBBR1KAN were obtained. The broad-host-range plasmid pLA2917 was transferred into Hyphomicrobium species by a triparental mating. However, this plasmid was integrated into the genome of Hyphomicrobium spp. Plasmids pLA2917, pKT231, pSUP2021, pRZ705, and phage DNA could not be transferred in Hyphomicrobium spp. by electroporation under the conditions applied.

  16. 3D Nanochannel Array Platform for High-throughput Cell Manipulation and Nano-electroporation

    Science.gov (United States)

    Chang, Lingqian

    Electroporation is one of the most common non-viral methods for gene delivery. Recent progress in gene therapy has offered special opportunities to electroporation for in vitro and in vivo applications. However, conventional bulk electroporation (BEP) inevitably causes serious cell damage and stochastic transfection between cells. Microfluidic electroporation (MEP) has been claimed to provide benign single cell transfection for the last decade. Nevertheless, the intracellular transport in both MEP and BEP systems is highly diffusion-dominant, which prevents precise dose control and high uniformity. In this Ph.D. research, we developed a 3D nanochannel-electroporation (3D NEP) platform for mass cell transfection. A silicon-based nanochannel array (3D NEP) chip was designed and fabricated for cell manipulation and electroporation. The chip, designed as Z-directional microchannel - nanochannel array, was fabricated by clean room techniques including projection photolithography and deep reactive-ion etching (DRIE). The fabricated 3D NEP chip is capable of handling 40,000 cells per 1 cm2, up to 1 million per wafer (100 mm diameter). High-throughput cell manipulation technologies were investigated for precise alignment of individual cells to the nanochannel array, a key step for NEP to achieve dose control. We developed three techniques for cell trapping in this work. (1) Magnetic tweezers (MTs) were integrated on the chip to remotely control cells under a programmed magnetic field. (2) A positive dielectrophoresis (pDEP) power system was built as an alternative to trap cells onto the nanochannel array using DEP force. (3) A novel yet simple 'dipping-trap' method was used to rapidly trap cells onto a nanochannel array, aligned by a micro-cap array pattern on the 3D NEP chip, which eventually offered 70 - 90 % trapping efficiency and 90 % specificity. 3D NEP platforms were assembled for cell transfection based on the Si-based nanochannel array chip and cell manipulation

  17. Single exponential decay waveform; a synergistic combination of electroporation and electrolysis (E2 for tissue ablation

    Directory of Open Access Journals (Sweden)

    Nina Klein

    2017-04-01

    Full Text Available Background Electrolytic ablation and electroporation based ablation are minimally invasive, non-thermal surgical technologies that employ electrical currents and electric fields to ablate undesirable cells in a volume of tissue. In this study, we explore the attributes of a new tissue ablation technology that simultaneously delivers a synergistic combination of electroporation and electrolysis (E2. Method A new device that delivers a controlled dose of electroporation field and electrolysis currents in the form of a single exponential decay waveform (EDW was applied to the pig liver, and the effect of various parameters on the extent of tissue ablation was examined with histology. Results Histological analysis shows that E2 delivered as EDW can produce tissue ablation in volumes of clinical significance, using electrical and temporal parameters which, if used in electroporation or electrolysis separately, cannot ablate the tissue. Discussion The E2 combination has advantages over the three basic technologies of non-thermal ablation: electrolytic ablation, electrochemical ablation (reversible electroporation with injection of drugs and irreversible electroporation. E2 ablates clinically relevant volumes of tissue in a shorter period of time than electrolysis and electroporation, without the need to inject drugs as in reversible electroporation or use paralyzing anesthesia as in irreversible electroporation.

  18. Overview of Single-Molecule Speckle (SiMS) Microscopy and Its Electroporation-Based Version with Efficient Labeling and Improved Spatiotemporal Resolution.

    Science.gov (United States)

    Yamashiro, Sawako; Watanabe, Naoki

    2017-07-06

    Live-cell single-molecule imaging was introduced more than a decade ago, and has provided critical information on remodeling of the actin cytoskeleton, the motion of plasma membrane proteins, and dynamics of molecular motor proteins. Actin remodeling has been the best target for this approach because actin and its associated proteins stop diffusing when assembled, allowing visualization of single-molecules of fluorescently-labeled proteins in a state specific manner. The approach based on this simple principle is called Single-Molecule Speckle (SiMS) microscopy. For instance, spatiotemporal regulation of actin polymerization and lifetime distribution of actin filaments can be monitored directly by tracking actin SiMS. In combination with fluorescently labeled probes of various actin regulators, SiMS microscopy has contributed to clarifying the processes underlying recycling, motion and remodeling of the live-cell actin network. Recently, we introduced an electroporation-based method called eSiMS microscopy, with high efficiency, easiness and improved spatiotemporal precision. In this review, we describe the application of live-cell single-molecule imaging to cellular actin dynamics and discuss the advantages of eSiMS microscopy over previous SiMS microscopy.

  19. Overview of Single-Molecule Speckle (SiMS Microscopy and Its Electroporation-Based Version with Efficient Labeling and Improved Spatiotemporal Resolution

    Directory of Open Access Journals (Sweden)

    Sawako Yamashiro

    2017-07-01

    Full Text Available Live-cell single-molecule imaging was introduced more than a decade ago, and has provided critical information on remodeling of the actin cytoskeleton, the motion of plasma membrane proteins, and dynamics of molecular motor proteins. Actin remodeling has been the best target for this approach because actin and its associated proteins stop diffusing when assembled, allowing visualization of single-molecules of fluorescently-labeled proteins in a state specific manner. The approach based on this simple principle is called Single-Molecule Speckle (SiMS microscopy. For instance, spatiotemporal regulation of actin polymerization and lifetime distribution of actin filaments can be monitored directly by tracking actin SiMS. In combination with fluorescently labeled probes of various actin regulators, SiMS microscopy has contributed to clarifying the processes underlying recycling, motion and remodeling of the live-cell actin network. Recently, we introduced an electroporation-based method called eSiMS microscopy, with high efficiency, easiness and improved spatiotemporal precision. In this review, we describe the application of live-cell single-molecule imaging to cellular actin dynamics and discuss the advantages of eSiMS microscopy over previous SiMS microscopy.

  20. Overview of Single-Molecule Speckle (SiMS) Microscopy and Its Electroporation-Based Version with Efficient Labeling and Improved Spatiotemporal Resolution

    Science.gov (United States)

    Yamashiro, Sawako; Watanabe, Naoki

    2017-01-01

    Live-cell single-molecule imaging was introduced more than a decade ago, and has provided critical information on remodeling of the actin cytoskeleton, the motion of plasma membrane proteins, and dynamics of molecular motor proteins. Actin remodeling has been the best target for this approach because actin and its associated proteins stop diffusing when assembled, allowing visualization of single-molecules of fluorescently-labeled proteins in a state specific manner. The approach based on this simple principle is called Single-Molecule Speckle (SiMS) microscopy. For instance, spatiotemporal regulation of actin polymerization and lifetime distribution of actin filaments can be monitored directly by tracking actin SiMS. In combination with fluorescently labeled probes of various actin regulators, SiMS microscopy has contributed to clarifying the processes underlying recycling, motion and remodeling of the live-cell actin network. Recently, we introduced an electroporation-based method called eSiMS microscopy, with high efficiency, easiness and improved spatiotemporal precision. In this review, we describe the application of live-cell single-molecule imaging to cellular actin dynamics and discuss the advantages of eSiMS microscopy over previous SiMS microscopy. PMID:28684722

  1. Can electroporation previous to radiofrequency hepatic ablation enlarge thermal lesion size? A feasibility study based on theoretical modelling and in vivo experiments.

    Science.gov (United States)

    Trujillo, Macarena; Castellví, Quim; Burdío, Fernando; Sánchez Velazquez, Patricia; Ivorra, Antoni; Andaluz, Anna; Berjano, Enrique

    2013-05-01

    The aim of this study was to assess the feasibility of a hybrid ablative technique based on applying electroporation (EP) pulses just before conducting radiofrequency ablation (RFA). The rationale was that the EP-induced reduction in blood perfusion could be sufficient to reduce the thermal sink effect and hence to increase the coagulation volume in comparison to that created exclusively by RFA. A modelling study and in vivo experimental study were used. A Cool-tip RF applicator was used both for EP and RFA. Overall, the results did not show any synergy effect from using the hybrid technique. Applying EP pulses prior to RFA did not increase the coagulation zone obtained and the lesions were almost identical. Additional computer simulations provided an explanation for this; the effect of reducing blood perfusion by thermal damage during RFA completely masks the effect of reducing blood perfusion by EP. This is because both thermal damage and EP affect the same zone, i.e. the tissue around the electrode. Our computer modelling and in vivo experimental findings suggest that the combination of EP and RFA with monopolar applicators does not provide an additional benefit over the use of RFA alone.

  2. Online bioimpedance feedback for in vivo electroporated tissues

    Science.gov (United States)

    Medrano, J.; Rey, J. I.; Connolly, R. J.; Anderson, A.; Jaroszeski, M.; Gitlin, R.

    2010-04-01

    Electroporation in vivo is a biotechnology method that uses short-duration high intensity electric fields to enhance plasma membrane permeability in living cells in order to facilitate the uptake of drugs, DNA, genes and proteins into the cytoplasm. The degree of permeability is related to the tissue's bioimpedance; hence, accurate impedance evaluation throughout electroporation treatment is essential to 1) avoid over-treating tissues resulting in excessive cell death and 2) under-treating tissues resulting in poor permeability. Cell viability and membrane permeability is based on a number of factors, including: time elapsed after electroporation, electroporation pulse amplitude, tissue type, and so on; thus, efficient feedback protocols must minimize delays between treatment and impedance readings. Current methods of bioimpedance feedback are often cumbersome and impedance analysis devices can be expensive, bulky, and immobile. Emerging technologies facilitate economical methods, fast protocols, and portability to realize bioimpedance measurement and feedback online (i.e. realtime). Consequently, this research uses automation software, logic-biased protocols, an inexpensive commercially available impedance analyzer microchip, and a custom-built hexagonal electrode probe to measure dynamic bioimpedance changes. This work demonstrates how this novel system measures tissue bioimpedance instantly and efficiently before and after electroporation. Additionally this system allows for the comparison of electrode geometries as well as electric field' magnitudes and distributions. Follow up work will pursue the optimization of plasma membrane permeability for several tissue/cell types.

  3. Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements.

    Directory of Open Access Journals (Sweden)

    Rosemary A Bamford

    Full Text Available Förster resonance energy transfer (FRET technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5 complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.

  4. Nanotechnology-based drug delivery systems

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    Singh Baljit

    2007-12-01

    Full Text Available Abstract Nanoparticles hold tremendous potential as an effective drug delivery system. In this review we discussed recent developments in nanotechnology for drug delivery. To overcome the problems of gene and drug delivery, nanotechnology has gained interest in recent years. Nanosystems with different compositions and biological properties have been extensively investigated for drug and gene delivery applications. To achieve efficient drug delivery it is important to understand the interactions of nanomaterials with the biological environment, targeting cell-surface receptors, drug release, multiple drug administration, stability of therapeutic agents and molecular mechanisms of cell signalling involved in pathobiology of the disease under consideration. Several anti-cancer drugs including paclitaxel, doxorubicin, 5-fluorouracil and dexamethasone have been successfully formulated using nanomaterials. Quantom dots, chitosan, Polylactic/glycolic acid (PLGA and PLGA-based nanoparticles have also been used for in vitro RNAi delivery. Brain cancer is one of the most difficult malignancies to detect and treat mainly because of the difficulty in getting imaging and therapeutic agents past the blood-brain barrier and into the brain. Anti-cancer drugs such as loperamide and doxorubicin bound to nanomaterials have been shown to cross the intact blood-brain barrier and released at therapeutic concentrations in the brain. The use of nanomaterials including peptide-based nanotubes to target the vascular endothelial growth factor (VEGF receptor and cell adhesion molecules like integrins, cadherins and selectins, is a new approach to control disease progression.

  5. A new equivalent circuit model for micro electroporation systems

    KAUST Repository

    Shagoshtasbi, Hooman

    2011-02-01

    Electroporation (EP) is a unique biotechnique in which intense electric pulses are applied on the cell membrane to temporarily generate nanoscale electropores and to increase the membrane permeability for the delivery of exogenous biomolecules or drugs. We propose a new equivalent circuit model with 8 electric components to predict the electrodynamic response of a micro EP system. As the permeability of the cell membrane increases, the membrane resistance decreases. The numerical simulations of the transmembrane current responses to different applied voltages (1∼6V) are consistent with the experimental results using HeLa cells. Besides, the transmembrane voltage as a function of applied voltages is determined as well. These transmembrane current and voltage responses can be extremely useful for the design of new generation of micro EP systems for transfection of large DNA molecules in the future. © 2011 IEEE.

  6. Protein-Based Drug-Delivery Materials

    Science.gov (United States)

    Jao, Dave; Xue, Ye; Medina, Jethro; Hu, Xiao

    2017-01-01

    There is a pressing need for long-term, controlled drug release for sustained treatment of chronic or persistent medical conditions and diseases. Guided drug delivery is difficult because therapeutic compounds need to survive numerous transport barriers and binding targets throughout the body. Nanoscale protein-based polymers are increasingly used for drug and vaccine delivery to cross these biological barriers and through blood circulation to their molecular site of action. Protein-based polymers compared to synthetic polymers have the advantages of good biocompatibility, biodegradability, environmental sustainability, cost effectiveness and availability. This review addresses the sources of protein-based polymers, compares the similarity and differences, and highlights characteristic properties and functionality of these protein materials for sustained and controlled drug release. Targeted drug delivery using highly functional multicomponent protein composites to guide active drugs to the site of interest will also be discussed. A systematical elucidation of drug-delivery efficiency in the case of molecular weight, particle size, shape, morphology, and porosity of materials will then be demonstrated to achieve increased drug absorption. Finally, several important biomedical applications of protein-based materials with drug-delivery function—including bone healing, antibiotic release, wound healing, and corneal regeneration, as well as diabetes, neuroinflammation and cancer treatments—are summarized at the end of this review. PMID:28772877

  7. Protein-Based Drug-Delivery Materials

    Directory of Open Access Journals (Sweden)

    Dave Jao

    2017-05-01

    Full Text Available There is a pressing need for long-term, controlled drug release for sustained treatment of chronic or persistent medical conditions and diseases. Guided drug delivery is difficult because therapeutic compounds need to survive numerous transport barriers and binding targets throughout the body. Nanoscale protein-based polymers are increasingly used for drug and vaccine delivery to cross these biological barriers and through blood circulation to their molecular site of action. Protein-based polymers compared to synthetic polymers have the advantages of good biocompatibility, biodegradability, environmental sustainability, cost effectiveness and availability. This review addresses the sources of protein-based polymers, compares the similarity and differences, and highlights characteristic properties and functionality of these protein materials for sustained and controlled drug release. Targeted drug delivery using highly functional multicomponent protein composites to guide active drugs to the site of interest will also be discussed. A systematical elucidation of drug-delivery efficiency in the case of molecular weight, particle size, shape, morphology, and porosity of materials will then be demonstrated to achieve increased drug absorption. Finally, several important biomedical applications of protein-based materials with drug-delivery function—including bone healing, antibiotic release, wound healing, and corneal regeneration, as well as diabetes, neuroinflammation and cancer treatments—are summarized at the end of this review.

  8. Multivalent human papillomavirus l1 DNA vaccination utilizing electroporation.

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    Kihyuck Kwak

    Full Text Available Naked DNA vaccines can be manufactured simply and are stable at ambient temperature, but require improved delivery technologies to boost immunogenicity. Here we explore in vivo electroporation for multivalent codon-optimized human papillomavirus (HPV L1 and L2 DNA vaccination.Balb/c mice were vaccinated three times at two week intervals with a fusion protein comprising L2 residues ∼11-88 of 8 different HPV types (11-88×8 or its DNA expression vector, DNA constructs expressing L1 only or L1+L2 of a single HPV type, or as a mixture of several high-risk HPV types and administered utilizing electroporation, i.m. injection or gene gun. Serum was collected two weeks and 3 months after the last vaccination. Sera from immunized mice were tested for in-vitro neutralization titer, and protective efficacy upon passive transfer to naive mice and vaginal HPV challenge. Heterotypic interactions between L1 proteins of HPV6, HPV16 and HPV18 in 293TT cells were tested by co-precipitation using type-specific monoclonal antibodies.Electroporation with L2 multimer DNA did not elicit detectable antibody titer, whereas DNA expressing L1 or L1+L2 induced L1-specific, type-restricted neutralizing antibodies, with titers approaching those induced by Gardasil. Co-expression of L2 neither augmented L1-specific responses nor induced L2-specific antibodies. Delivery of HPV L1 DNA via in vivo electroporation produces a stronger antibody response compared to i.m. injection or i.d. ballistic delivery via gene gun. Reduced neutralizing antibody titers were observed for certain types when vaccinating with a mixture of L1 (or L1+L2 vectors of multiple HPV types, likely resulting from heterotypic L1 interactions observed in co-immunoprecipitation studies. High titers were restored by vaccinating with individual constructs at different sites, or partially recovered by co-expression of L2, such that durable protective antibody titers were achieved for each type

  9. Generating Mouse Models Using Zygote Electroporation of Nucleases (ZEN) Technology with High Efficiency and Throughput.

    Science.gov (United States)

    Wang, Wenbo; Zhang, Yingfan; Wang, Haoyi

    2017-01-01

    Mouse models with genetic modifications are widely used in biology and biomedical research. Although the application of CRISPR-Cas9 system greatly accelerated the process of generating genetically modified mice, the delivery method depending on manual injection of the components into the embryos remains a bottleneck, as it is laborious, low throughput, and technically demanding. To overcome this limitation, we invented and optimized the ZEN (Zygote electroporation of nucleases) technology to deliver CRISPR-Cas9 reagents via electroporation. Using ZEN, we were able to generate genetically modified mouse models with high efficiency and throughput. Here, we describe the protocol in great detail.

  10. Polysaccharide-Based Micelles for Drug Delivery

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    Nan Zhang

    2013-05-01

    Full Text Available Delivery of hydrophobic molecules and proteins has been an issue due to poor bioavailability following administration. Thus, micelle carrier systems are being investigated to improve drug solubility and stability. Due to problems with toxicity and immunogenicity, natural polysaccharides are being explored as substitutes for synthetic polymers in the development of new micelle systems. By grafting hydrophobic moieties to the polysaccharide backbone, self-assembled micelles can be readily formed in aqueous solution. Many polysaccharides also possess inherent bioactivity that can facilitate mucoadhesion, enhanced targeting of specific tissues, and a reduction in the inflammatory response. Furthermore, the hydrophilic nature of some polysaccharides can be exploited to enhance circulatory stability. This review will highlight the advantages of polysaccharide use in the development of drug delivery systems and will provide an overview of the polysaccharide-based micelles that have been developed to date.

  11. Nanochemistry of protein-based delivery agents

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    Subin R.C.K. Rajendran

    2016-07-01

    Full Text Available The past decade has seen an increased interest in the conversion of food proteins into functional biomaterials, including their use for loading and delivery of physiologically active compounds such as nutraceuticals and pharmaceuticals. Proteins possess a competitive advantage over other platforms for the development of nanodelivery systems since they are biocompatible, amphipathic, and widely available. Proteins also have unique molecular structures and diverse functional groups that can be selectively modified to alter encapsulation and release properties. A number of physical and chemical methods have been used for preparing protein nanoformulations, each based on different underlying protein chemistry. This review focuses on the chemistry of the reorganization and/or modification of proteins into functional nanostructures for delivery, from the perspective of their preparation, functionality, stability and physiological behavior.

  12. Local electroporation of a single cell using a scanning ion conductance microscope

    Science.gov (United States)

    Iwata, Futoshi; Yamazaki, Koji; Ishizaki, Kimihiro; Ushiki, Tatuo

    2014-03-01

    We developed a novel electroporation technique for molecular delivery into a single cell. A nanopipette, a thermally pulled glass capillary, is prepared as to act as a pair of tiny electrodes for single-cell electroporation. An Ag/AgCl wire is inserted into the nanopipette, and the outside edge of the nanopipette is coated by Ag sputtering. Electric pulses are applied between the outside and inside electrodes to form a local electric field at the edge of the nanopipette. To position the pipette edge in the vicinity of the cell membrane, we control the probe-surface distance using a scanning ion conductance microscope (SICM). The SICM technique achieves non-contact approach of the nanopipette edge on the cell membrane, which allows low-invasive electroporation of a single cell. As a demonstration of this technique, a fluorescent molecule of propidium iodide was successfully delivered into a single HeLa cell.

  13. Electroporation of cells using EM induction of ac fields by a magnetic stimulator

    Energy Technology Data Exchange (ETDEWEB)

    Chen, C; Robinson, M P [Department of Electronics, University of York, Heslington, York YO10 5DD (United Kingdom); Evans, J A [Academic Unit of Medical Physics, University of Leeds, Leeds LS2 9JT (United Kingdom); Smye, S W [Department of Medical Physics and Engineering, Leeds Teaching Hospitals, St. James' s University Hospital, Leeds LS9 7TF (United Kingdom); O' Toole, P [Department of Biology, University of York, Heslington, York YO10 5DD (United Kingdom)

    2010-02-21

    This paper describes a method of effectively electroporating mammalian cell membranes with pulsed alternating-current (ac) electric fields at field strengths of 30-160 kV m{sup -1}. Although many in vivo electroporation protocols entail applying square wave or monotonically decreasing pulses via needles or electrode plates, relatively few have explored the use of pulsed ac fields. Following our previous study, which established the effectiveness of ac fields for electroporating cell membranes, a primary/secondary coil system was constructed to produce sufficiently strong electric fields by electromagnetic induction. The primary coil was formed from the applicator of an established transcranial magnetic stimulation (TMS) system, while the secondary coil was a purpose-built device of a design which could eventually be implanted into tissue. The effects of field strength, pulse interval and cumulative exposure time were investigated using microscopy and flow cytometry. Results from experiments on concentrated cell suspensions showed an optimized electroporation efficiency of around 50%, demonstrating that electroporation can be practicably achieved by inducing such pulsed ac fields. This finding confirms the possibility of a wide range of in vivo applications based on magnetically coupled ac electroporation.

  14. Retinal Fiber Tracing by In Ovo Electroporation

    Science.gov (United States)

    Harada, Hidekiyo; Nakamura, Harukazu

    Axonal tracing techniques are the fundamentals for the investigation of neural circuit formation. In ovo electroporation system allows us to transfect a gene of interest to the desired place in chick embryos (Odani et al., 2008). Recently, Tol2 transposase element, which was originally found in medaka fish (Koga et al., 1996), has been adapted to an in ovo electroporation system (Niwa et al., 1991; Kawakami et al., 1998, 2000, 2004a, 2004b; Kawakami & Noda, 2004; Kawakami, 2005, 2007; Sato et al., 2007). This system assures the integration of the transgene into the genome by electroporation (Niwa et al., 1991; Sato et al., 2007). We applied this system for tracing retinal fibers (Harada et al., 2008). In this chapter, we demonstrate the method of tracing retinal fibers from both small and large groups of the retinal ganglion cell (RGC) with transposon-mediated gene transfer by in ovo electroporation to chick embryos.

  15. Biopolymer based nanosystem for doxorubicin targeted delivery.

    Science.gov (United States)

    Csikós, Zsuzsanna; Kerekes, Krisztina; Fazekas, Erika; Kun, Sándor; Borbély, János

    2017-01-01

    This study describes formation of an actively and passively targeted, water-soluble drug delivery system (DDS) which contains doxorubicin (DOX). The system comprises two biocompatible and biodegradable polymers: poly-γ-glutamic acid (PGA) and chitosan (CH). Self-assembly of these biopolymers in aqueous medium results stable nanoparticles (NPs) with a hydrodynamic size of 80-150 nm and slightly negative surface charge. Folic acid (FA) was used as targeting agent bonded to the polyanion (PA) and also to the surface of the NPs. The NP's physical stability, active targeting effect, cellular toxicity, release profile and in vivo anti-tumor efficacy were investigated. It was found that the targeted, self-assembled nanoparticles are stable at 4°C for several months, cause better in vitro toxicity effect on folate receptor (FR) positive cell lines than the doxorubicin or the non-targeted nanosystem and based on its release profile it is expected, that the nanosystem will remain stable during the circulation in the body. Pharmacodynamic studies demonstrated that the DOX-loaded nanoparticles can deliver greater tumor growth inhibition than the free drug molecules and the liposomal compound, with less general toxicity. It was observed that the overall survival is the main benefit of the biopolymer based drug delivery system.

  16. Controlled release of optimized electroporation enhances the transdermal efficiency of sinomenine hydrochloride for treating arthritis in vitro and in clinic

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    Feng S

    2017-06-01

    Full Text Available Shun Feng,1,* Lijun Zhu,1,* Zhisheng Huang,2 Haojia Wang,1 Hong Li,1 Hua Zhou,3 Linlin Lu,1 Ying Wang,1 Zhongqiu Liu,1,3 Liang Liu1,3 1International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 2Department of Acupuncture and Rehabilitation, Guangzhou Hospital of Integrated Traditional Chinese and Western Medicine, Guangzhou, Guangdong, 3State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, People’s Republic of China *These authors contributed equally to this work Abstract: Sinomenine hydrochloride (SH is an ideal drug for the treatment of rheumatoid arthritis and osteoarthritis. However, high plasma concentration of systemically administered SH can release histamine, which can cause rash and gastrointestinal side effects. Topical delivery can increase SH concentration in the synovial fluid without high plasma level, thus minimizing systemic side effects. However, passive diffusion of SH was found to be inefficient because of the presence of the stratum corneum layer. Therefore, an effective method is required to compensate for the low efficiency of SH passive diffusion. In this study, transdermal experiments in vitro and clinical tests were utilized to explore the optimized parameters for electroporation of topical delivery for SH. Fluorescence experiment and hematoxylin and eosin staining analysis were performed to reveal the mechanism by which electroporation promoted permeation. In vitro, optimized electroporation parameters were 3 KHz, exponential waveform, and intensity 10. Using these parameters, transdermal permeation of SH was increased by 1.9–10.1 fold in mice skin and by 1.6–47.1 fold in miniature pig skin compared with passive diffusion. After the electroporation stimulation, the intercellular intervals and epidermal cracks in the skin increased. In clinical tests, SH concentration in synovial fluid was 20

  17. In vivo Editing of the Human Mutant Rhodopsin Gene by Electroporation of Plasmid-based CRISPR/Cas9 in the Mouse Retina

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    Maria Carmela Latella

    2016-01-01

    Full Text Available The bacterial CRISPR/Cas system has proven to be an efficient tool for genetic manipulation in various organisms. Here we show the application of CRISPR-Cas9 technology to edit the human Rhodopsin (RHO gene in a mouse model for autosomal dominant Retinitis Pigmentosa. We designed single or double sgRNAs to knock-down mutant RHO expression by targeting exon 1 of the RHO gene carrying the P23H dominant mutation. By delivering Cas9 and sgRNAs in a single plasmid we induced an efficient gene editing in vitro, in HeLa cells engineered to constitutively express the P23H mutant RHO allele. Similarly, after subretinal electroporation of the CRISPR/Cas9 plasmid expressing two sgRNAs into P23H RHO transgenic mice, we scored specific gene editing as well as significant reduction of the mutant RHO protein. Successful in vivo application of the CRISPR/Cas9 system confirms its efficacy as a genetic engineering tool in photoreceptor cells.

  18. In vivo Editing of the Human Mutant Rhodopsin Gene by Electroporation of Plasmid-based CRISPR/Cas9 in the Mouse Retina.

    Science.gov (United States)

    Latella, Maria Carmela; Di Salvo, Maria Teresa; Cocchiarella, Fabienne; Benati, Daniela; Grisendi, Giulia; Comitato, Antonella; Marigo, Valeria; Recchia, Alessandra

    2016-11-22

    The bacterial CRISPR/Cas system has proven to be an efficient tool for genetic manipulation in various organisms. Here we show the application of CRISPR-Cas9 technology to edit the human Rhodopsin (RHO) gene in a mouse model for autosomal dominant Retinitis Pigmentosa. We designed single or double sgRNAs to knock-down mutant RHO expression by targeting exon 1 of the RHO gene carrying the P23H dominant mutation. By delivering Cas9 and sgRNAs in a single plasmid we induced an efficient gene editing in vitro, in HeLa cells engineered to constitutively express the P23H mutant RHO allele. Similarly, after subretinal electroporation of the CRISPR/Cas9 plasmid expressing two sgRNAs into P23H RHO transgenic mice, we scored specific gene editing as well as significant reduction of the mutant RHO protein. Successful in vivo application of the CRISPR/Cas9 system confirms its efficacy as a genetic engineering tool in photoreceptor cells.

  19. Optimised electroporation mediated DNA vaccination for treatment of prostate cancer.

    LENUS (Irish Health Repository)

    Ahmad, Sarfraz

    2010-01-01

    ABSTRACT: BACKGROUND: Immunological therapies enhance the ability of the immune system to recognise and destroy cancer cells via selective killing mechanisms. DNA vaccines have potential to activate the immune system against specific antigens, with accompanying potent immunological adjuvant effects from unmethylated CpG motifs as on prokaryotic DNA. We investigated an electroporation driven plasmid DNA vaccination strategy in animal models for treatment of prostate cancer. METHODS: Plasmid expressing human PSA gene (phPSA) was delivered in vivo by intra-muscular electroporation, to induce effective anti-tumour immune responses against prostate antigen expressing tumours. Groups of male C57 BL\\/6 mice received intra-muscular injections of phPSA plasmid. For phPSA delivery, quadriceps muscle was injected with 50 mug plasmid. After 80 seconds, square-wave pulses were administered in sequence using a custom designed pulse generator and acustom-designed applicator with 2 needles placed through the skin central to the muscle. To determine an optimum treatment regimen, three different vaccination schedules were investigated. In a separate experiment, the immune potential of the phPSA vaccine was further enhanced with co- administration of synthetic CpG rich oligonucleotides. One week after last vaccination, the mice were challenged subcutaneously with TRAMPC1\\/hPSA (prostate cancer cell line stably expressing human PSA) and tumour growth was monitored. Serum from animals was examined by ELISA for anti-hPSA antibodies and for IFNgamma. Histological assessment of the tumours was also carried out. In vivo and in vitro cytotoxicity assays were performed with splenocytes from treated mice. RESULTS: The phPSA vaccine therapy significantly delayed the appearance of tumours and resulted in prolonged survival of the animals. Four-dose vaccination regimen provided optimal immunological effects. Co - administration of the synthetic CpG with phPSA increased anti-tumour responses

  20. CRY 1AB trangenic cowpea obtained by nodal electroporation ...

    African Journals Online (AJOL)

    Electroporation-mediated genetic transformation was used to introduce Cry 1 Ab insecticidal gene into cowpea. Nodal buds were electroporated in planta with a plasmid carrying the Cry 1Ab and antibiotic resistance npt II genes driven by a 35S CaMV promoter. T1 seeds derived from electroporated branches were selected ...

  1. Protein-Based Nanomedicine Platforms for Drug Delivery

    Energy Technology Data Exchange (ETDEWEB)

    Ma Ham, Aihui; Tang, Zhiwen; Wu, Hong; Wang, Jun; Lin, Yuehe

    2009-08-03

    Drug delivery systems have been developed for many years, however some limitations still hurdle the pace of going to clinical phase, for example, poor biodistribution, drug molecule cytotoxicity, tissue damage, quick clearance from the circulation system, solubility and stability of drug molecules. To overcome the limitations of drug delivery, biomaterials have to be developed and applied to drug delivery to protect the drug molecules and to enhance the drug’s efficacy. Protein-based nanomedicine platforms for drug delivery are platforms comprised of naturally self-assembled protein subunits of the same protein or a combination of proteins making up a complete system. They are ideal for drug delivery platforms due to their biocompatibility and biodegradability coupled with low toxicity. A variety of proteins have been used and characterized for drug delivery systems including the ferritin/apoferritin protein cage, plant derived viral capsids, the small Heat shock protein (sHsp) cage, albumin, soy and whey protein, collagen, and gelatin. There are many different types and shapes that have been prepared to deliver drug molecules using protein-based platforms including the various protein cages, microspheres, nanoparticles, hydrogels, films, minirods and minipellets. There are over 30 therapeutic compounds that have been investigated with protein-based drug delivery platforms for the potential treatment of various cancers, infectious diseases, chronic diseases, autoimmune diseases. In protein-based drug delivery platforms, protein cage is the most newly developed biomaterials for drug delivery and therapeutic applications. Their uniform sizes, multifunctions, and biodegradability push them to the frontier for drug delivery. In this review, the recent strategic development of drug delivery has been discussed with a special emphasis upon the polymer based, especially protein-based nanomedicine platforms for drug delivery. The advantages and disadvantages are also

  2. Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle

    DEFF Research Database (Denmark)

    Hjortkjær, Camilla Brolin; Shiraishi, Takehiko; Hojman, Pernille

    2015-01-01

    Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality for impro......Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality...... and dystrophic mdx mice with or without electroporation. At low, single PNA doses (1.5, 3, or 10 µg/TA), electroporation augmented the antisense exon skipping induced by an unmodified PNA by twofold to fourfold in healthy mouse muscle with optimized electric parameters, measured after 7 days. The PNA splice...

  3. Clinician attendance and delivery practices at hospital-based vaginal deliveries in Western Kenya.

    Science.gov (United States)

    Patel, Suha J; Bogaert, Kelly; Gachuno, Onesmus W; Kibore, Minnie W; Unger, Jennifer; Walker, Dilys M

    2016-05-01

    To characterize delivery practices and factors associated with respectful, evidence-based care at a referral hospital in Western Kenya. An exploratory observational study used a standardized birth-observation form to record information on patient characteristics and healthcare practitioner behaviors during uncomplicated vaginal deliveries between June 30, 2014 and July 17, 2014. All deliveries were monitored for whether healthcare staff performed six specific evidence-based practices (three maternal and three neonatal practices). In total, 75 vaginal deliveries were observed. In 48 (64%) deliveries, nursing students were the only practitioners present. The mean number of evidence-based practices performed at each delivery was 3.58. The number of evidence-based practices performed by junior practitioners was higher when a nurse educator was assessing their performance (4.47 vs 3.36, Prespectful-care scores were recorded when delivery teams comprised three or more practitioners (1.38; 95% confidence interval 0.93-1.84 vs 2.74; 95% confidence interval 2.16-3.31, P=0.002). The present study found low rates of evidence-based practice and respectful maternity care; this could serve as a deterrent for women seeking care at the study facility. These findings emphasize the need for a comprehensive approach in increasing the quality of patient care to improve maternal and newborn health outcomes. Copyright © 2015 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

  4. Dextran-based microspheres as controlled delivery systems for proteins

    NARCIS (Netherlands)

    Vlugt-Wensink, K.D.F.

    2007-01-01

    Dextran-based microspheres as controlled delivery systems for proteins Dextran based microspheres are investigated as controlled delivery system for proteins. Microspheres were prepared by polymerization of dex-HEMA in an aqueous two-phase system of dex-HEMA and PEG. Protein loaded microspheres are

  5. Transformation of group A streptococci by electroporation

    NARCIS (Netherlands)

    Suvorov, Alexander; Kok, Jan; Venema, Gerhardus

    1988-01-01

    The introduction, via electroporation, of free plasmid DNA into three strains of Streptococcus pyogenes is described. The method is very simple and rapid and efficiencies vary from 1 × 10^3 to 4 × 10^4 per µg of DNA. The method was also used to introduce an integrative plasmid and transformants were

  6. Theoretical and experimental study of electroporation of red blood cells using MEMS technology

    KAUST Repository

    Deng, Peigang

    2010-01-01

    A theoretical and experimental study of electroporation (EP) of red blood cells (RBCs) was presented in this paper. With additional strain energy, an energy-based model of an electropore induced on a RBC\\'s membrane at different electric fields was proposed to predict the critical EP electric field strength. In addition, EP experiments with red blood cells at single-cell level was carried out on a micro EP chip. The measured critical EP electric field strengths are in agreement with the numerical predictions. ©2010 IEEE.

  7. Superparamagnetic Iron Oxide Nanoparticle-Based Delivery Systems for Biotherapeutics

    Science.gov (United States)

    Mok, Hyejung; Zhang, Miqin

    2014-01-01

    Introduction Superparamagnetic iron oxide nanoparticle (SPION)-based carrier systems have many advantages over other nanoparticle-based systems. They are biocompatible, biodegradable, facilely tunable, and superparamagnetic and thus controllable by an external magnetic field. These attributes enable their broad biomedical applications. In particular, magnetically-driven carriers are drawing considerable interest as an emerging therapeutic delivery system because of their superior delivery efficiency. Area covered This article reviews the recent advances in use of SPION-based carrier systems to improve the delivery efficiency and target specificity of biotherapeutics. We examine various formulations of SPION-based delivery systems, including SPION micelles, clusters, hydrogels, liposomes, and micro/nanospheres, as well as their specific applications in delivery of biotherapeutics. Expert opinion Recently, biotherapeutics including therapeutic cells, proteins and genes have been studied as alternative treatments to various diseases. Despite the advantages of high target specificity and low adverse effects, clinical translation of biotherapeutics has been hindered by the poor stability and low delivery efficiency compared to chemical drugs. Accordingly, biotherapeutic delivery systems that can overcome these limitations are actively pursued. SPION-based materials can be ideal candidates for developing such delivery systems because of their excellent biocompatibility and superparamagnetism that enables long-term accumulation/retention at target sites by utilization of a suitable magnet. In addition, synthesis technologies for production of finely-tuned, homogeneous SPIONs have been well developed, which may promise their rapid clinical translation. PMID:23199200

  8. The site of administration influences both the type and the magnitude of the immune response induced by DNA vaccine electroporation.

    Science.gov (United States)

    Vandermeulen, Gaëlle; Vanvarenberg, Kevin; De Beuckelaer, Ans; De Koker, Stefaan; Lambricht, Laure; Uyttenhove, Catherine; Reschner, Anca; Vanderplasschen, Alain; Grooten, Johan; Préat, Véronique

    2015-06-22

    We investigated the influence of the site of administration of DNA vaccine on the induced immune response. DNA vaccines were administered by electroporation at three different sites: tibial cranial muscle, abdominal skin and ear pinna. Aiming to draw general conclusions about DNA vaccine delivery, we successively used several plasmids encoding either luciferase and ovalbumin as models or gp160 and P1A as vaccines against HIV and P815 mastocytoma, respectively. Low levels and duration of luciferase transgene expression were observed after electroporation of the abdominal skin, partly explaining its lower immunogenic performance as compared to the other sites of administration. Analyses of OT-I CD8+ and OT-II CD4+ T cell responses highlighted the differential impact of the delivery site on the elicited immune response. Muscle electroporation induced the strongest humoral immune response and both muscle and ear pinna sites induced cellular immunity against gp160. Ear pinna delivery generated the highest level of CTL responses against P1A but electroporation of muscle and ear pinna were equally efficient in delaying P815 growth and improving mice survival. The present study demonstrated that the site of administration is a key factor to be tested in the development of DNA vaccine. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Self-Nanoemulsifying Drug Delivery Systems Based on Melon Oil ...

    African Journals Online (AJOL)

    Purpose: To formulate self-nanoemulsifying drug delivery systems (SNEDDS) based on melon oil and its admixture with a homolipid from Bos indicus (cow fat) for the delivery of indomethacin, a hydrophobic anti-inflammatory agent. Method: Melon oil and cow fat were extracted by standard methods and used in the ...

  10. An ICT-Based Agricultural Extension Service Delivery for Nigeria ...

    African Journals Online (AJOL)

    This paper proposed an ICT-based extension service delivery for Nigeria. The proposed design, though to be use as supplement to the existing system would engender an extension delivery system that is void of many of the limitations inherent in the earlier approaches. Basically, it revolves round the use ICT facilities like ...

  11. Recent developments in oral lipid-based drug delivery

    DEFF Research Database (Denmark)

    Thomas, N.; Rades, T.; Müllertz, A.

    2013-01-01

    The increasing number of poorly water-soluble drugs in development in the pharmaceutical industry has sparked interest in novel drug delivery options such as lipid-based drug delivery systems (LbDDS). Several LbDDS have been marketed successfully and have shown superior and more reliable...

  12. Lightning-triggered electroporation and electrofusion as possible contributors to natural horizontal gene transfer

    Science.gov (United States)

    Kotnik, Tadej

    2013-09-01

    Phylogenetic studies show that horizontal gene transfer (HGT) is a significant contributor to genetic variability of prokaryotes, and was perhaps even more abundant during the early evolution. Hitherto, research of natural HGT has mainly focused on three mechanisms of DNA transfer: conjugation, natural competence, and viral transduction. This paper discusses the feasibility of a fourth such mechanism - cell electroporation and/or electrofusion triggered by atmospheric electrostatic discharges (lightnings). A description of electroporation as a phenomenon is followed by a review of experimental evidence that electroporation of prokaryotes in aqueous environments can result in release of non-denatured DNA, as well as uptake of DNA from the surroundings and transformation. Similarly, a description of electrofusion is followed by a review of experiments showing that prokaryotes devoid of cell wall can electrofuse into hybrids expressing the genes of their both precursors. Under sufficiently fine-tuned conditions, electroporation and electrofusion are efficient tools for artificial transformation and hybridization, respectively, but the quantitative analysis developed here shows that conditions for electroporation-based DNA release, DNA uptake and transformation, as well as for electrofusion are also present in many natural aqueous environments exposed to lightnings. Electroporation is thus a plausible contributor to natural HGT among prokaryotes, and could have been particularly important during the early evolution, when the other mechanisms might have been scarcer or nonexistent. In modern prokaryotes, natural absence of the cell wall is rare, but it is reasonable to assume that the wall has formed during a certain stage of evolution, and at least prior to this, electrofusion could also have contributed to natural HGT. The concluding section outlines several guidelines for assessment of the feasibility of lightning-triggered HGT.

  13. Nanotechnologies in delivery of mRNA therapeutics using nonviral vector-based delivery systems.

    Science.gov (United States)

    Guan, S; Rosenecker, J

    2017-03-01

    Because of its safe and effective protein expression profile, in vitro transcribed messenger RNA (IVT-mRNA) represents a promising candidate in the development of novel therapeutics for genetic diseases, vaccines or gene editing strategies, especially when its inherent shortcomings (for example, instability and immunogenicity) have been partially addressed via structural modifications. However, numerous unsolved technical difficulties in successful in vivo delivery of IVT-mRNA have greatly hindered the applications of IVT-mRNA in clinical development. Recent advances in nanotechnology and material science have yielded many promising nonviral delivery systems, some of which were able to efficiently facilitate targeted in vivo delivery of IVT-mRNA in safe and noninvasive manners. The diversity and flexibility of these delivery systems highlight the recent progress of IVT-mRNA-based therapy using nonviral vectors. In this review, we summarize recent advances of existing and emerging nonviral vector-based nanotechnologies for IVT-mRNA delivery and briefly summarize the interesting but rarely discussed applications on simultaneous delivery of IVT-mRNA with DNA.

  14. Calcium electroporation in three cell lines; a comparison of bleomycin and calcium, calcium compounds, and pulsing conditions

    DEFF Research Database (Denmark)

    Frandsen, Stine Krog; Gissel, Hanne; Hojman, Pernille

    2013-01-01

    BACKGROUND: Electroporation with calcium (calcium electroporation) can induce ATP depletion-associated cellular death. In the clinical setting, the cytotoxic drug bleomycin is currently used with electroporation (electrochemotherapy) for palliative treatment of tumors. Calcium electroporation off...

  15. Oral delivery of peptides and proteins using lipid-based drug delivery systems

    DEFF Research Database (Denmark)

    Li, Ping; Nielsen, Hanne Mørck; Müllertz, Anette

    2012-01-01

    INTRODUCTION: In order to successfully develop lipid-based drug delivery systems (DDS) for oral administration of peptides and proteins, it is important to gain an understanding of the colloid structures formed by these DDS, the mode of peptide and protein incorporation as well as the mechanism...... by which intestinal absorption of peptides and proteins is promoted. AREAS COVERED: The present paper reviews the literature on lipid-based DDS, employed for oral delivery of peptides and proteins and highlights the mechanisms by which the different lipid-based carriers are expected to overcome the two...... most important barriers (extensive enzymatic degradation and poor transmucosal permeability). This paper also gives a clear-cut idea about advantages and drawbacks of using different lipidic colloidal carriers ((micro)emulsions, solid lipid core particles and liposomes) for oral delivery of peptides...

  16. In vivo imaging of cancer cells with electroporation of quantum dots and multispectral imaging

    Science.gov (United States)

    Yoo, Jung Sun; Won, Nayoun; Kim, Hong Bae; Bang, Jiwon; Kim, Sungjee; Ahn, Saeyoung; Soh, Kwang-Sup

    2010-06-01

    Our understanding of dissemination and growth of cancer cells is limited by our inability for long-term followup of this process in vivo. Fluorescence molecular imaging has the potential to track cancer cells with high contrast and sensitivity in living animals. For this purpose, intracellular delivery of near-infrared fluorescence quantum dots (QDs) by electroporation offers considerable advantages over organic fluorophores and other cell tagging methods. In this research we developed a multispectral imaging system that could eliminate two major parameters compromising in vivo fluorescence imaging performance, i.e., variations in the tissue optical properties and tissue autofluorescence. We demonstrated that electroporation of QDs and multispectral imaging allowed in vivo assessment of cancer development and progression in the xenograft mouse tumor model for more than 1 month, providing a powerful means to learn more about the biology of cancer and metastasis.

  17. Dendrimer based nanotherapeutics for ocular drug delivery

    Science.gov (United States)

    Kambhampati, Siva Pramodh

    PAMAM dendrimers are a class of well-defined, hyperbranched polymeric nanocarriers that are being investigated for ocular drug and gene delivery. Their favorable properties such as small size, multivalency and water solubility can provide significant opportunities for many biologically unstable drugs and allows potentially favorable ocular biodistribution. This work exploits hydroxyl terminated dendrimers (G4-OH) as drug/gene delivery vehicles that can target retinal microglia and pigment epithelium via systemic delivery with improved efficacy at much lower concentrations without any side effects. Two different drugs Triamcinolone acetonide (TA) and N-Acetyl Cysteine (NAC) conjugated to G4-OH dendrimers showed tailorable sustained release in physiological relevant solutions and were evaluated in-vitro and in-vivo. Dendrimer-TA conjugates enhanced the solubility of TA and were 100 fold more effective at lower concentrations than free TA in its anti-inflammatory activity in activated microglia and in suppressing VEGF production in hypoxic RPE cells. Dendrimers targeted activated microglia/macrophages and RPE and retained for a period of 21 days in I/R mice model. The relative retention of intravitreal and intravenous dendrimers was comparable, if a 30-fold intravenous dose is used; suggesting intravenous route targeting retinal diseases are possible with dendrimers. D-NAC when injected intravenously attenuated retinal and choroidal inflammation, significantly reduced (˜73%) CNV growth at early stage of AMD in rat model of CNV. A combination therapy of D-NAC + D-TA significantly suppressed microglial activation and promoted CNV regression in late stages of AMD without causing side-effects. G4-OH was modified with linker having minimal amine groups and incorporation of TA as a nuclear localization enhancer resulted in compact gene vectors with favorable safety profile and achieved high levels of transgene expression in hard to transfect human retinal pigment

  18. Carbohydrate-based amphiphilic nano delivery systems for cancer therapy.

    Science.gov (United States)

    Liu, Kegang; Jiang, Xiaohua; Hunziker, Patrick

    2016-09-15

    Nanoparticles (NPs) are novel drug delivery systems that have been attracting more and more attention in recent years, and have been used for the treatment of cancer, infection, inflammation and other diseases. Among the numerous classes of materials employed for constructing NPs, organic polymers are outstanding due to the flexibility of design and synthesis and the ease of modification and functionalization. In particular, NP based amphiphilic polymers make a great contribution to the delivery of poorly-water soluble drugs. For example, natural, biocompatible and biodegradable products like polysaccharides are widely used as building blocks for the preparation of such drug delivery vehicles. This review will detail carbohydrate based amphiphilic polymeric systems for cancer therapy. Specifically, it focuses on the nature of the polymer employed for the preparation of targeted nanocarriers, the synthetic methods, as well as strategies for the application and evaluation of biological activity. Applications of the amphiphilic polymer systems include drug delivery, gene delivery, photosensitizer delivery, diagnostic imaging and specific ligand-assisted cellular uptake. As a result, a thorough understanding of the relationship between chemical structure and biological properties facilitate the optimal design and rational clinical application of the resulting carbohydrate based nano delivery systems for cancer therapy.

  19. Functional genomics tool: Gene silencing in Ixodes scapularis eggs and nymphs by electroporated dsRNA

    Directory of Open Access Journals (Sweden)

    Troiano Emily

    2010-01-01

    Full Text Available Abstract Background Ticks are blood-sucking arthropods responsible for transmitting a wide variety of disease-causing agents, and constitute important public health threats globally. Ixodes scapularis is the primary vector of the Lyme disease agent in the eastern and central U.S. RNAi is a mechanism by which gene-specific double-stranded RNA (dsRNA triggers degradation of homologous mRNA transcripts. Here, we describe an optimized protocol for effectively suppressing gene expression in the egg and nymphal stages of I. scapularis by electroporation. Results The genes encoding the putative Phospholipase A2 (PLA2, cytoplasmic Cystatin, Syntaxin-5, β-Actin and Calreticulin were targeted by delivering the dsRNA encoding the specific gene coding regions in the unfed nymphs. Silencing was measured using real time qRT-PCR. Electroporation as a mode of dsRNA delivery appears to be substantially efficient and less traumatic to the tick than dsRNA microinjection in the unfed nymphs. Using Cy3-labeled dsRNA to monitor the movement, electroporated dsRNA entered the nymphs and spread to salivary glands and other tissues. The significant disruption of β-actin and cytoplasmic Cystatin transcripts in tick eggs demonstrate the applicability of this technique. The PLA2, cytoplasmic Cystatin, Syntaxin-5, β-Actin and Calreticulin genes were also significantly silenced, suggesting that this method has the potential to introduce dsRNA in eggs and unfed nymphs. Conclusions Our study demonstrates that electroporation can be used as a simple dsRNA delivery tool in assessing the functional role of tick genes in the vector-host interactions. This technique represents a novel approach for specific gene suppression in immature stages of ticks.

  20. Chitosan-Based Multifunctional Platforms for Local Delivery of Therapeutics

    Directory of Open Access Journals (Sweden)

    Seong-Chul Hong

    2017-03-01

    Full Text Available Chitosan has been widely used as a key biomaterial for the development of drug delivery systems intended to be administered via oral and parenteral routes. In particular, chitosan-based microparticles are the most frequently employed delivery system, along with specialized systems such as hydrogels, nanoparticles and thin films. Based on the progress made in chitosan-based drug delivery systems, the usefulness of chitosan has further expanded to anti-cancer chemoembolization, tissue engineering, and stem cell research. For instance, chitosan has been used to develop embolic materials designed to efficiently occlude the blood vessels by which the oxygen and nutrients are supplied. Indeed, it has been reported to be a promising embolic material. For better anti-cancer effect, embolic materials that can locally release anti-cancer drugs were proposed. In addition, a complex of radioactive materials and chitosan to be locally injected into the liver has been investigated as an efficient therapeutic tool for hepatocellular carcinoma. In line with this, a number of attempts have been explored to use chitosan-based carriers for the delivery of various agents, especially to the site of interest. Thus, in this work, studies where chitosan-based drug delivery systems have successfully been used for local delivery will be presented along with future perspectives.

  1. Combining Electrolysis and Electroporation for Tissue Ablation.

    Science.gov (United States)

    Phillips, Mary; Rubinsky, Liel; Meir, Arie; Raju, Narayan; Rubinsky, Boris

    2015-08-01

    Electrolytic ablation is a method that operates by delivering low magnitude direct current to the target region over long periods of time, generating electrolytic products that destroy cells. This study was designed to explore the hypothesis stating that electrolytic ablation can be made more effective when the electrolysis-producing electric charges are delivered using electric pulses with field strength typical in reversible electroporation protocols. (For brevity we will refer to tissue ablation protocols that combine electroporation and electrolysis as E(2).) The mechanistic explanation of this hypothesis is related to the idea that products of electrolysis generated by E(2) protocols can gain access to the interior of the cell through the electroporation permeabilized cell membrane and therefore cause more effective cell death than from the exterior of an intact cell. The goal of this study is to provide a first-order examination of this hypothesis by comparing the charge dosage required to cause a comparable level of damage to a rat liver, in vivo, when using either conventional electrolysis or E(2) approaches. Our results show that E(2) protocols produce tissue damage that is consistent with electrolytic ablation. Furthermore, E(2) protocols cause damage comparable to that produced by conventional electrolytic protocols while delivering orders of magnitude less charge to the target tissue over much shorter periods of time. © The Author(s) 2014.

  2. Electropore Formation in Mechanically Constrained Phospholipid Bilayers.

    Science.gov (United States)

    Fernández, M Laura; Risk, Marcelo Raúl; Vernier, P Thomas

    2017-11-23

    Molecular dynamics simulations of lipid bilayers in aqueous systems reveal how an applied electric field stabilizes the reorganization of the water-membrane interface into water-filled, membrane-spanning, conductive pores with a symmetric, toroidal geometry. The pore formation process and the resulting symmetric structures are consistent with other mathematical approaches such as continuum models formulated to describe the electroporation process. Some experimental data suggest, however, that the shape of lipid electropores in living cell membranes may be asymmetric. We describe here the axially asymmetric pores that form when mechanical constraints are applied to selected phospholipid atoms. Electropore formation proceeds even with severe constraints in place, but pore shape and pore formation time are affected. Since lateral and transverse movement of phospholipids may be restricted in cell membranes by covalent attachments to or non-covalent associations with other components of the membrane or to membrane-proximate intracellular or extracellular biomolecular assemblies, these lipid-constrained molecular models point the way to more realistic representations of cell membranes in electric fields.

  3. Hydrocolloid-based nutraceutical delivery systems.

    Science.gov (United States)

    Janaswamy, Srinivas; Youngren, Susanne R

    2012-05-01

    Nutraceuticals are important due to their inherent health benefits. However, utilization and consumption are limited by their poor water solubility and instability at normal processing and storage conditions. Herein, we propose an elegant and novel approach for the delivery of nutraceuticals in their active form using hydrocolloid matrices that are inexpensive and non-toxic with generally recognized as safe (GRAS) status. Iota-carrageenan and curcumin have been chosen as models of hydrocolloid and nutraceutical compounds, respectively. The iota-carrageenan network maintains a stable organization after encapsulating curcumin molecules, protects them from melting and then releases them in a sustained manner. These findings lay a strong foundation for developing value-added functional and medicinal foods.

  4. Hydrocolloid-based nutraceutical delivery systems

    Energy Technology Data Exchange (ETDEWEB)

    Janaswamy, Srinivas; Youngren, Susanne R. (Purdue)

    2012-07-11

    Nutraceuticals are important due to their inherent health benefits. However, utilization and consumption are limited by their poor water solubility and instability at normal processing and storage conditions. Herein, we propose an elegant and novel approach for the delivery of nutraceuticals in their active form using hydrocolloid matrices that are inexpensive and non-toxic with generally recognized as safe (GRAS) status. Iota-carrageenan and curcumin have been chosen as models of hydrocolloid and nutraceutical compounds, respectively. The iota-carrageenan network maintains a stable organization after encapsulating curcumin molecules, protects them from melting and then releases them in a sustained manner. These findings lay a strong foundation for developing value-added functional and medicinal foods.

  5. Nanoparticle-based drug delivery to the vagina: a review

    Science.gov (United States)

    Ensign, Laura M.; Cone, Richard; Hanes, Justin

    2014-01-01

    Vaginal drug administration can improve prophylaxis and treatment of many conditions affecting the female reproductive tract, including sexually transmitted diseases, fungal and bacterial infections, and cancer. However, achieving sustained local drug concentrations in the vagina can be challenging, due to the high permeability of the vaginal epithelium and expulsion of conventional soluble drug dosage forms. Nanoparticle-based drug delivery platforms have received considerable attention for vaginal drug delivery, as nanoparticles can provide sustained release, cellular targeting, and even intrinsic antimicrobial or adjuvant properties that can improve the potency and/or efficacy of prophylactic and therapeutic modalities. Here, we review the use of polymeric nanoparticles, liposomes, dendrimers, and inorganic nanoparticles for vaginal drug delivery. Although most of the work toward nanoparticle-based drug delivery in the vagina has been focused on HIV prevention, strategies for treatment and prevention of other sexually transmitted infections, treatment for reproductive tract cancer, and treatment of fungal and bacterial infections are also highlighted. PMID:24830303

  6. siRNA delivery with lipid-based systems

    DEFF Research Database (Denmark)

    Foged, Camilla

    2012-01-01

    A key hurdle for the further development of RNA interference (RNAi) therapeutics like small interfering RNA (siRNA) is their safe and effective delivery. Lipids are promising and versatile carriers because they are based on Nature's own building blocks and can be provided with properties which...... allow for protection of the siRNA, steric stabilization, targeting, membrane fusion and triggered drug release. At present a variety of lipid-based transfectants for siRNA delivery have been used for in vitro and in vivo purposes. The majority bears a cationic charge to electrostatically complex the siRNA...... into more hydrophobic lipoplexes, which promote passage of the siRNA across cellular membrane barriers, especially when lipids are added that facilitate membrane fusion. Despite these attractive features, siRNA delivery vehicles are facing a number of challenges such as the limited delivery efficiency...

  7. Delivery Strategies for Stem Cell-Based Therapy

    Directory of Open Access Journals (Sweden)

    Jason P. Glotzbach

    2012-01-01

    Full Text Available Before stem cell-based therapies can become a clinical reality, technologies for cell delivery must be developed that can control differentiation and pluripotency, maintain a hospitable environment for cell survival and function, and provide a structural framework for regenerative healing of the target tissue. Insights gained from developmental and stem cell biology should guide the design of devices and techniques to facilitate stem cell-based therapies. Several strategies have been developed for surgical delivery of stem cells, including synthetic and biologic matrices for cell seeding, complex biochemical delivery devices for maintenance and modulation of stem cell properties, and smart constructs with the ability to adapt to the dynamic in vivo environment after implantation. In aggregate, surgical delivery of complex stem cell-seeded constructs has the potential to revolutionize surgical therapies for a wide range of diseases in order to provide a more regenerative platform for tissue and organ healing.

  8. Functional evaluation of malaria Pfs25 DNA vaccine by in vivo electroporation in olive baboons.

    Science.gov (United States)

    Kumar, Rajesh; Nyakundi, Ruth; Kariuki, Thomas; Ozwara, Hastings; Nyamongo, Onkoba; Mlambo, Godfree; Ellefsen, Barry; Hannaman, Drew; Kumar, Nirbhay

    2013-06-28

    Plasmodium falciparum Pfs25 antigen, expressed on the surface of zygotes and ookinetes, is one of the leading targets for the development of a malaria transmission-blocking vaccine (TBV). Our laboratory has been evaluating DNA plasmid based Pfs25 vaccine in mice and non-human primates. Previously, we established that in vivo electroporation (EP) delivery is an effective method to improve the immunogenicity of DNA vaccine encoding Pfs25 in mice. In order to optimize the in vivo EP procedure and test for its efficacy in more clinically relevant larger animal models, we employed in vivo EP to evaluate the immune response and protective efficacy of Pfs25 encoding DNA vaccine in nonhuman primates (olive baboons, Papio anubis). The results showed that at a dose of 2.5mg DNA vaccine, antibody responses were significantly enhanced with EP as compared to without EP resulting in effective transmission blocking efficiency. Similar immunogenicity enhancing effect of EP was also observed with lower doses (0.5mg and 1mg) of DNA plasmids. Further, final boosting with a single dose of recombinant Pfs25 protein resulted in dramatically enhanced antibody titers and significantly increased functional transmission blocking efficiency. Our study suggests priming with DNA vaccine via EP along with protein boost regimen as an effective method to elicit potent immunogenicity of malaria DNA vaccines in nonhuman primates and provides the basis for further evaluation in human volunteers. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Polycation-Based Ternary Gene Delivery System.

    Science.gov (United States)

    Liu, Shuai; Guo, Tianying

    2015-01-01

    Recent progress in gene therapy has opened the door for various human diseases. The greatest challenge that gene vectors still face is the ability to sufficiently deliver nucleic acid into target cells. To overcome various barriers, plenty of researches have been undertaken utilizing diverse strategies, among which a wide variety of polycation/pDNA vectors have been developed and explored frequently. For enhanced transfection efficiency, polycations are constantly utilized with covalent modifications, which however lead to reduced positive charge density and changed properties of polycation/pDNA complexes. Accordingly, non-covalent or ternary strategy is proposed. The cationic properties of polycations can be retained and the transfection efficiency can be enhanced by introducing additional polymers with functional groups via non-covalent assembly. This review will discuss the construction and advantages of ternary complexes gene delivery system, including low toxicity and enhanced gene expression both in vitro and in vivo. Recent progress and expectations with promising results that may have some reference for clinical application are also discussed.

  10. Emulgel based topical delivery system for loratadine

    Directory of Open Access Journals (Sweden)

    Vijay Kumar

    2015-01-01

    Full Text Available The present study was taken up with the objective of formulating emulgels to facilitate topical delivery of loratadine in the treatment of localized skin allergy. It was attempted to prepare the emulgels using three different types of surfactants, i.e. cationic, non-ionic and anionic.   Further, the aim was to compare the formulations for their drug release and stability. Loratadine was incorporated into an o/w system, which was subsequently gellified using Carbopol 940. The resulting emulgels were subject to tests for physical characteristics. Thereafter, ex vivo drug release study, skin irritation test and in vivo test for anti-allergic activity were also carried out.  A considerably higher drug release was recorded from the emulgel formulated with the cationic surfactant, cetrimide. Drug release kinetics were investigated by fitting the drug release data into various models. The drug release from all the formulations was found to follow zero-order kinetics. In addition, the prepared emulgels exhibited satisfactory physical characteristics and good stability. Besides being non-irritant to the skin, they were effective in alleviating symptoms of skin allergy.

  11. Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation

    Directory of Open Access Journals (Sweden)

    Xiaojun Liu

    2017-05-01

    Full Text Available ABSTRACT The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB. The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.

  12. Polymeric Biomaterial and Lipid Based Nanoparticles for Oral Drug Delivery.

    Science.gov (United States)

    Dilnawaz, Fahima

    2017-01-01

    Oral drug delivery is widespread owing to its non-invasive nature which complements high patient compliance. However, the drug administration via oral route is quite challenging due to the presence of the biochemical barriers which hinders the uptake as well as access to blood stream. Apart from that, stability, poor solubility and bioavailability of administered drug via the gastrointestinal (GI) tract are also exigent. Till now various oral formulations were developed which releases the drug in a timely manner but lacks appropriate therapeutic concentration. Recently nanoparticles based drug delivery system has emerged as prominent strategy for optimizing the oral drug delivery and maximizing the treatment efficiency. Besides, different strategic polymeric nanoparticles are engineered for interaction both at extracellular and intracellular levels with gastrointestinal mucosa. The review article focuses on the polymeric and lipid based various nanocarriers that have been widely studied for the enhanced oral drug delivery of different therapeutic molecules and addresses recent progress of biocompatible and biodegradable polymeric nanoparticles approach for its improvement. The progress of numerous oral nanoparticulate drug delivery vehicles will be immensely helpful to improve therapeutic efficacy with reduced adverse side effects. Unlike other forms of administration, it will have better patient compliance and soothing effect. The oral drug delivery will certainly play a pivotal role soon in expanding the clinical repertoire and applications. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. Effects of Circular DNA Length on Transfection Efficiency by Electroporation into HeLa Cells.

    Science.gov (United States)

    Hornstein, Benjamin D; Roman, Dany; Arévalo-Soliz, Lirio M; Engevik, Melinda A; Zechiedrich, Lynn

    2016-01-01

    The ability to produce extremely small and circular supercoiled vectors has opened new territory for improving non-viral gene therapy vectors. In this work, we compared transfection of supercoiled DNA vectors ranging from 383 to 4,548 bp, each encoding shRNA against GFP under control of the H1 promoter. We assessed knockdown of GFP by electroporation into HeLa cells. All of our vectors entered cells in comparable numbers when electroporated with equal moles of DNA. Despite similar cell entry, we found length-dependent differences in how efficiently the vectors knocked down GFP. As vector length increased up to 1,869 bp, GFP knockdown efficiency per mole of transfected DNA increased. From 1,869 to 4,257 bp, GFP knockdown efficiency per mole was steady, then decreased with increasing vector length. In comparing GFP knockdown with equal masses of vectors, we found that the shorter vectors transfect more efficiently per nanogram of DNA transfected. Our results rule out cell entry and DNA mass as determining factors for gene knockdown efficiency via electroporation. The length-dependent effects we have uncovered are likely explained by differences in nuclear translocation or transcription. These data add an important step towards clinical applications of non-viral vector delivery.

  14. Transfer of foreign DNA into aquatic animals by electroporation

    Science.gov (United States)

    This chapter describes the principle, procedure and application of the electroporation method to produce various types of transgenic marine organisms including finfish, shellfish and marine algae. Electroporation utilizes a series of short electrical pulses to induce formation of short-lived pores ...

  15. Malaria treatment using novel nano-based drug delivery systems.

    Science.gov (United States)

    Baruah, Uday Krishna; Gowthamarajan, Kuppusamy; Vanka, Ravisankar; Karri, Veera Venkata Satyanarayana Reddy; Selvaraj, Kousalya; Jojo, Gifty M

    2017-08-01

    We reside in an era of technological innovation and advancement despite which infectious diseases like malaria remain to be one of the greatest threats to the humans. Mortality rate caused by malaria disease is a huge concern in the twenty-first century. Multiple drug resistance and nonspecific drug targeting of the most widely used drugs are the main reasons/drawbacks behind the failure in malarial therapy. Dose-related toxicity because of high doses is also a major concern. Therefore, to overcome these problems nano-based drug delivery systems are being developed to facilitate site-specific or target-based drug delivery and hence minimizing the development of resistance progress and dose-dependent toxicity issues. In this review, we discuss about the shortcomings in treating malaria and how nano-based drug delivery systems can help in curtailing the infectious disease malaria.

  16. Whole tumor antigen vaccination using dendritic cells: Comparison of RNA electroporation and pulsing with UV-irradiated tumor cells

    Directory of Open Access Journals (Sweden)

    Benencia Fabian

    2008-04-01

    Full Text Available Abstract Because of the lack of full characterization of tumor associated antigens for solid tumors, whole antigen use is a convenient approach to tumor vaccination. Tumor RNA and apoptotic tumor cells have been used as a source of whole tumor antigen to prepare dendritic cell (DC based tumor vaccines, but their efficacy has not been directly compared. Here we compare directly RNA electroporation and pulsing of DCs with whole tumor cells killed by ultraviolet (UV B radiation using a convenient tumor model expressing human papilloma virus (HPV E6 and E7 oncogenes. Although both approaches led to DCs presenting tumor antigen, electroporation with tumor cell total RNA induced a significantly higher frequency of tumor-reactive IFN-gamma secreting T cells, and E7-specific CD8+ lymphocytes compared to pulsing with UV-irradiated tumor cells. DCs electroporated with tumor cell RNA induced a larger tumor infiltration by T cells and produced a significantly stronger delay in tumor growth compared to DCs pulsed with UV-irradiated tumor cells. We conclude that electroporation with whole tumor cell RNA and pulsing with UV-irradiated tumor cells are both effective in eliciting antitumor immune response, but RNA electroporation results in more potent tumor vaccination under the examined experimental conditions.

  17. Corona discharge in electroporation of cell membranes

    Energy Technology Data Exchange (ETDEWEB)

    Cramariuc, R; Nisiparu, L [Competence Centre in Electrostatics and Electrotehchnologies (Romania); Tudorache, A; Branduse, E; Fotescu, L [Research Institute of Wine Processing, Valea Mantei Street, No.l, Valea Calugareasca (Romania); Popa, M E; Mitelut, A [Biotechnology Faculty, University of Agronomical Sciences and Veterinary Medicine (Romania); Turtoi, M O

    2008-12-01

    The objective of the present work is to demonstrate that electrical corona discharge is very efficient in cellular membrane electroporation due to current pulses with sharp front (2-5 ns) and to the fact that corona discharge is associated with UV radiation and micro particles emission. A comparison between DC and AC at 800 Hz and a special waveform to corona application is presented. The comparison is analyzed by means of applying all these in the maceration process (electroplasmolysis) of red wine production and in the processes of different types of the microbes.

  18. Layered Double Hydroxide-Based Nanocarriers for Drug Delivery

    Directory of Open Access Journals (Sweden)

    Xue Bi

    2014-06-01

    Full Text Available Biocompatible clay materials have attracted particular attention as the efficient drug delivery systems (DDS. In this article, we review developments in the use of layered double hydroxides (LDHs for controlled drug release and delivery. We show how advances in the ability to synthesize intercalated structures have a significant influence on the development of new applications of these materials. We also show how modification and/or functionalization can lead to new biotechnological and biomedical applications. This review highlights the most recent progresses in research on LDH-based controlled drug delivery systems, focusing mainly on: (i DDS with cardiovascular drugs as guests; (ii DDS with anti-inflammatory drugs as guests; and (iii DDS with anti-cancer drugs as guests. Finally, future prospects for LDH-based drug carriers are also discussed.

  19. Skin electroporation: effects on transgene expression, DNA persistence and local tissue environment

    DEFF Research Database (Denmark)

    Roos, Anna-Karin; Eriksson, Fredrik; Timmons, James A

    2009-01-01

    . METHODOLOGY/PRINCIPAL FINDINGS: This study investigates intradermal DNA electrovaccination in detail and describes the effects on expression of the vaccine antigen, plasmid persistence and the local tissue environment. Gene profiling of the vaccination site showed that the combination of DNA......-administered, PSA-specific T cells were induced and concurrently the luciferase expression became undetectable. Electroporation did not affect the long-term persistence of the PSA-expressing plasmid. CONCLUSIONS/SIGNIFICANCE: This study provides important insights to how DNA delivery by intradermal...

  20. How to move towards community based service delivery?

    NARCIS (Netherlands)

    Meuwissen, L.; Voorham, T.; Bakker, D. de

    2007-01-01

    Aim: Community based primary health care offers in potential the opportunity to tailor health service delivery to the needs and demands of the local population. Up to now, there is no clear cut method to do this. In a pilot benchmark for general practices, data were collected on demand and

  1. Value-Based Delivery of Education: MOOCs as Messengers

    Science.gov (United States)

    Gilfoil, David M.; Focht, Jeffrey W.

    2015-01-01

    Value-based delivery of healthcare has been discussed in the literature for almost a decade. The concept focuses on the patient and defines value as the improvement of patient outcomes divided by healthcare costs. Further refinements, called the Triple Aim model, focus on improving patient outcomes, reducing treatment costs, and improving patient…

  2. The Relationship between Facility-Based Delivery and Infant ...

    African Journals Online (AJOL)

    AJRH Managing Editor

    Valentino Manase Lema. Breast Cancer and Sexual Dysfunction. African Journal of Reproductive Health June 2016; 20 (2): 27. ORIGINAL RESEARCH ARTICLE. The Relationship between Facility-Based Delivery and Infant. Immunization in sub-Saharan Africa. Cheryl A. Moyer. 1,2,3. , Dana Benyas. 1 and Sarah Rominski.

  3. Cationic solid-lipid nanoparticles are as efficient as electroporation in DNA vaccination against visceral leishmaniasis in mice.

    Science.gov (United States)

    Saljoughian, N; Zahedifard, F; Doroud, D; Doustdari, F; Vasei, M; Papadopoulou, B; Rafati, S

    2013-12-01

    The use of an appropriate delivery system has recently emerged as a promising approach for the development of effective vaccination against visceral leishmaniasis (VL). Here, we compare two vaccine delivery systems, namely electroporation and cationic solid-lipid nanoparticle (cSLN) formulation, to administer a DNA vaccine harbouring the L. donovani A2 antigen along with L. infantum cysteine proteinases [CPA and CPB without its unusual C-terminal extension (CPB(-CTE) )] and evaluate their potential against L. infantum challenge. Prime-boost administration of the pcDNA-A2-CPA-CPB(-CTE) delivered by either electroporation or cSLN formulation protects BALB/c mice against L. infantum challenge and that protective immunity is associated with high levels of IFN-γ and lower levels of IL-10 production, leading to a strong Th1 immune response. At all time points, the ratio of IFN-γ: IL-10 induced upon restimulation with rA2-rCPA-rCPB and F/T antigens was significantly higher in vaccinated animals. Moreover, Th2-efficient protection was elicited through a high humoral immune response. Nitric oxide production, parasite burden and histopathological analysis were also in concordance with other findings. Overall, these data indicate that similar to the electroporation delivery system, cSLNs as a nanoscale vehicle of Leishmania antigens could improve immune response, hence indicating the promise of these strategies against visceral leishmaniasis. © 2013 John Wiley & Sons Ltd.

  4. Targeted Drug Delivery System for Platinum-based Anticancer Drugs.

    Science.gov (United States)

    Gao, Chuanzhu; Zhang, Yan; Chen, Ji; Wang, Tianshuai; Qian, Yunxu; Yang, Bo; Dong, Peng; Zhang, Yingjie

    2016-01-01

    Platinum-based (Pt-based) anticancer drugs have been recognised as one of the most effective drugs for clinical treatment of malignant tumors due to its unique mechanism of action and broad range of anticancer spectrum. But, there are still some limitations such as side effects, drug resistance/cross resistance, no-specific targeting, becoming obstacles to restrict its expanding of clinical application. Targeted drug delivery system (TDDS) is a promising strategy for the research of novel Pt-based anticancer drugs. A variety of TDDS have been explored to improve the antitumor activity of Pt-based drugs such as nanoparticle drug systems, polymer-drug systems, drugs-macrocyclic compounds systems, etc. The review concentrates on recent development of various targeted drug delivery techniques, which could provide more opportunities for the development of Pt-based drugs with better efficiency, lower toxicity and less resistance.

  5. Efficient expression of transgenes in adult zebrafish by electroporation

    Directory of Open Access Journals (Sweden)

    Rao S Hari

    2005-10-01

    Full Text Available Abstract Background Expression of transgenes in muscle by injection of naked DNA is widely practiced. Application of electrical pulses at the site of injection was demonstrated to improve transgene expression in muscle tissue. Zebrafish is a precious model to investigate developmental biology in vertebrates. In this study we investigated the effect of electroporation on expression of transgenes in 3–6 month old adult zebrafish. Results Electroporation parameters such as number of pulses, voltage and amount of plasmid DNA were optimized and it was found that 6 pulses of 40 V·cm-1 at 15 μg of plasmid DNA per fish increased the luciferase expression 10-fold compared to controls. Similar enhancement in transgene expression was also observed in Indian carp (Labeo rohita. To establish the utility of adult zebrafish as a system for transient transfections, the strength of the promoters was compared in A2 cells and adult zebrafish after electroporation. The relative strengths of the promoters were found to be similar in cell lines and in adult zebrafish. GFP fluorescence in tissues after electroporation was also studied by fluorescence microscopy. Conclusion Electroporation after DNA injection enhances gene expression 10-fold in adult zebrafish. Electroporation parameters for optimum transfection of adult zebrafish with tweezer type electrode were presented. Enhanced reporter gene expression upon electroporation allowed comparison of strengths of the promoters in vivo in zebrafish.

  6. Nanoparticle-based drug delivery to the vagina: a review.

    Science.gov (United States)

    Ensign, Laura M; Cone, Richard; Hanes, Justin

    2014-09-28

    Vaginal drug administration can improve prophylaxis and treatment of many conditions affecting the female reproductive tract, including sexually transmitted diseases, fungal and bacterial infections, and cancer. However, achieving sustained local drug concentrations in the vagina can be challenging, due to the high permeability of the vaginal epithelium and expulsion of conventional soluble drug dosage forms. Nanoparticle-based drug delivery platforms have received considerable attention for vaginal drug delivery, as nanoparticles can provide sustained release, cellular targeting, and even intrinsic antimicrobial or adjuvant properties that can improve the potency and/or efficacy of prophylactic and therapeutic modalities. Here, we review the use of polymeric nanoparticles, liposomes, dendrimers, and inorganic nanoparticles for vaginal drug delivery. Although most of the work toward nanoparticle-based drug delivery in the vagina has been focused on HIV prevention, strategies for treatment and prevention of other sexually transmitted infections, treatment for reproductive tract cancer, and treatment of fungal and bacterial infections are also highlighted. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Electroporation enhances antimicrobial photodynamic therapy mediated by the hydrophobic photosensitizer, hypericin, Electroporation enhances antimicrobial photodynamic inactivation

    OpenAIRE

    de Melo, Wanessa de Cássia Martins Antunes; Lee, Alexander N; Perussi, Janice Rodrigues; Hamblin, Michael R.

    2013-01-01

    The effective transport of photosensitizers (PS) across the membrane and the intracellular accumulation of PS are the most crucial elements in antimicrobial photodynamic therapy (aPDT). However, due to the morphological complexity of Gram-negative bacteria the penetration of PS is limited, especially hydrophobic PS. Electroporation (EP) could increase the effectiveness of aPDT, by promoting the formation of transient pores that enhance the permeability of the bacterial membrane to PS. In this...

  8. Electroporation increases antitumoral efficacy of the bcl-2 antisense G3139 and chemotherapy in a human melanoma xenograft

    Directory of Open Access Journals (Sweden)

    Baldi Alfonso

    2011-07-01

    Full Text Available Abstract Background Nucleic acids designed to modulate the expression of target proteins remain a promising therapeutic strategy in several diseases, including cancer. However, clinical success is limited by the lack of efficient intracellular delivery. In this study we evaluated whether electroporation could increase the delivery of antisense oligodeoxynucleotides against bcl-2 (G3139 as well as the efficacy of combination chemotherapy in human melanoma xenografts. Methods Melanoma-bearing nude mice were treated i.v. with G3139 and/or cisplatin (DDP followed by the application of trains of electric pulses to tumors. Western blot, immunohistochemistry and real-time PCR were performed to analyze protein and mRNA expression. The effect of electroporation on muscles was determined by histology, while tumor apoptosis and the proliferation index were analyzed by immunohistochemistry. Antisense oligodeoxynucleotides tumor accumulation was measured by FACS and confocal microscopy. Results The G3139/Electroporation combined therapy produced a significant inhibition of tumor growth (TWI, more than 50% accompanied by a marked tumor re-growth delay (TRD, about 20 days. The efficacy of this treatment was due to the higher G3139 uptake in tumor cells which led to a marked down-regulation of bcl-2 protein expression. Moreover, the G3139/EP combination treatment resulted in an enhanced apoptotic index and a decreased proliferation rate of tumors. Finally, an increased tumor response was observed after treatment with the triple combination G3139/DDP/EP, showing a TWI of about 75% and TRD of 30 days. Conclusions These results demonstrate that electroporation is an effective strategy to improve the delivery of antisense oligodeoxynucleotides within tumor cells in vivo and it may be instrumental in optimizing the response of melanoma to chemotherapy. The high response rate observed in this study suggest to apply this strategy for the treatment of melanoma patients.

  9. Mapping of bionic array electric field focusing in plasmid DNA-based gene electrotransfer.

    Science.gov (United States)

    Browne, C J; Pinyon, J L; Housley, D M; Crawford, E N; Lovell, N H; Klugmann, M; Housley, G D

    2016-04-01

    Molecular medicine through gene therapy is challenged to achieve targeted action. This is now possible utilizing bionic electrode arrays for focal delivery of naked (plasmid) DNA via gene electrotransfer. Here, we establish the properties of array-based electroporation affecting targeted gene delivery. An array with eight 300 μm platinum ring electrodes configured as a cochlear implant bionic interface was used to transduce HEK293 cell monolayers with a plasmid-DNA green fluorescent protein (GFP) reporter gene construct. Electroporation parameters were pulse intensity, number, duration, separation and electrode configuration. The latter determined the shape of the electric fields, which were mapped using a voltage probe. Electrode array-based electroporation was found to require ~100 × lower applied voltages for cell transduction than conventional electroporation. This was found to be due to compression of the field lines orthogonal to the array. A circular area of GFP-positive cells was created when the electrodes were ganged together as four adjacent anodes and four cathodes, whereas alternating electrode polarity created a linear area of GFP-positive cells. The refinement of gene delivery parameters was validated in vivo in the guinea pig cochlea. These findings have significant clinical ramifications, where spatiotemporal control of gene expression can be predicted by manipulation of the electric field via current steering at a cellular level.

  10. A nonlinear electromechanical coupling model for electropore expansion in cell electroporation

    KAUST Repository

    Deng, Peigang

    2014-10-15

    Under an electric field, the electric tractions acting on a cell membrane containing a pore-nucleus are investigated by using a nonlinear electromechanical coupling model, in which the cell membrane is treated as a hyperelastic material. Iterations between the electric field and the structure field are performed to reveal the electrical forces exerting on the pore region and the subsequent pore expansion process. An explicit exponential decay of the membrane\\'s edge energy as a function of pore radius is defined for a hydrophilic pore and the transition energy as a hydrophobic pore converts to a hydrophilic pore during the initial stage of pore formation is investigated. It is found that the edge energy for the creation of an electropore edge plays an important role at the atomistic scale and it determines the hydrophobic-hydrophilic transition energy barrier. Various free energy evolution paths are exhibited, depending on the applied electric field, which provides further insight towards the electroporation (EP) phenomenon. In comparison with previous EP models, the proposed model has the ability to predict the metastable point on the free energy curve that is relevant to the lipid ion channel. In addition, the proposed model can also predict the critical transmembrane potential for the activation of an effective electroporation that is in a good agreement with previously published experimental data.

  11. Anistropically varying conductivity in irreversible electroporation simulations.

    Science.gov (United States)

    Labarbera, Nicholas; Drapaca, Corina

    2017-11-01

    One recent area of cancer research is irreversible electroporation (IRE). Irreversible electroporation is a minimally invasive procedure where needle electrodes are inserted into the body to ablate tumor cells with electricity. The aim of this paper is to propose a mathematical model that incorporates a tissue's conductivity increasing more in the direction of the electrical field as this has been shown to occur in experiments. It was necessary to mathematically derive a valid form of the conductivity tensor such that it is dependent on the electrical field direction and can be easily implemented into numerical software. The derivation of a conductivity tensor that can take arbitrary functions for the conductivity in the directions tangent and normal to the electrical field is the main contribution of this paper. Numerical simulations were performed for isotropic-varying and anisotropic-varying conductivities to evaluate the importance of including the electrical field's direction in the formulation for conductivity. By starting from previously published experimental results, this paper derived a general formulation for an anistropic-varying tensor for implementation into irreversible electroporation modeling software. The anistropic-varying tensor formulation allows the conductivity to take into consideration both electrical field direction and magnitude, as opposed to previous published works that only took into account electrical field magnitude. The anisotropic formulation predicts roughly a five percent decrease in ablation size for the monopolar simulation and approximately a ten percent decrease in ablation size for the bipolar simulations. This is a positive result as previously reported results found the isotropic formulation to overpredict ablation size for both monopolar and bipolar simulations. Furthermore, it was also reported that the isotropic formulation overpredicts the ablation size more for the bipolar case than the monopolar case. Thus, our

  12. Dual delivery systems based on polyamine analog BENSpm as prodrug and gene delivery vectors

    Science.gov (United States)

    Zhu, Yu

    Combination drug and gene therapy shows promise in cancer treatment. However, the success of such strategy requires careful selection of the therapeutic agents, as well as development of efficient delivery vectors. BENSpm (N 1, N11-bisethylnorspermine), a polyamine analogue targeting the intracellular polyamine pathway, draws our special attention because of the following reasons: (1) polyamine pathway is frequently dysregulated in cancer; (2) BENSpm exhibits multiple functions to interfere with the polyamine pathway, such as to up-regulate polyamine metabolism enzymes and down-regulate polyamine biosynthesis enzymes. Therefore BENSpm depletes all natural polyamines and leads to apoptosis and cell growth inhibition in a wide range of cancers; (3) preclinical studies proved that BENSpm can act synergistically with various chemotherapy agents, making it a promising candidate in combination therapy; (4) multiple positive charges in BENSpm enable it as a suitable building block for cationic polymers, which can be further applied to gene delivery. In this dissertation, our goal was to design dual-function delivery vector based on BENSpm that can function as a gene delivery vector and, after intracellular degradation, as an active anticancer agent targeting dysregulated polyamine metabolism. We first demonstrated strong synergism between BENSpm and a potential therapeutic gene product TRAIL. Strong synergism was obtained in both estrogen-dependent MCF-7 breast cancer cells and triple-negative MDA-MB-231 breast cancer cells. Significant dose reduction of TRAIL in combination with BENSpm in MDA-MB-231 cells, together with the fact that BENSpm rendered MCF-7 cells more sensitive to TRAIL treatment verified our rationale of designing BENSpm-based delivery platform. This was expected to be beneficial for overcoming drug resistance in chemotherapy, as well as boosting the therapeutic effect of therapeutic genes. We first designed a lipid-based BENSpm dual vector (Lipo

  13. Carrier-Based Drug Delivery System for Treatment of Acne

    Science.gov (United States)

    Vyas, Amber; Kumar Sonker, Avinesh

    2014-01-01

    Approximately 95% of the population suffers at some point in their lifetime from acne vulgaris. Acne is a multifactorial disease of the pilosebaceous unit. This inflammatory skin disorder is most common in adolescents but also affects neonates, prepubescent children, and adults. Topical conventional systems are associated with various side effects. Novel drug delivery systems have been used to reduce the side effect of drugs commonly used in the topical treatment of acne. Topical treatment of acne with active pharmaceutical ingredients (API) makes direct contact with the target site before entering the systemic circulation which reduces the systemic side effect of the parenteral or oral administration of drug. The objective of the present review is to discuss the conventional delivery systems available for acne, their drawbacks, and limitations. The advantages, disadvantages, and outcome of using various carrier-based delivery systems like liposomes, niosomes, solid lipid nanoparticles, and so forth, are explained. This paper emphasizes approaches to overcome the drawbacks and limitations associated with the conventional system and the advances and application that are poised to further enhance the efficacy of topical acne formulations, offering the possibility of simplified dosing regimen that may improve treatment outcomes using novel delivery system. PMID:24688376

  14. Chitosan nanoparticle based delivery systems for sustainable agriculture.

    Science.gov (United States)

    Kashyap, Prem Lal; Xiang, Xu; Heiden, Patricia

    2015-01-01

    Development of technologies that improve food productivity without any adverse impact on the ecosystem is the need of hour. In this context, development of controlled delivery systems for slow and sustained release of agrochemicals or genetic materials is crucial. Chitosan has emerged as a valuable carrier for controlled delivery of agrochemicals and genetic materials because of its proven biocompatibility, biodegradability, non-toxicity, and adsorption abilities. The major advantages of encapsulating agrochemicals and genetic material in a chitosan matrix include its ability to function as a protective reservoir for the active ingredients, protecting the ingredients from the surrounding environment while they are in the chitosan domain, and then controlling their release, allowing them to serve as efficient gene delivery systems for plant transformation or controlled release of pesticides. Despite the great progress in the use of chitosan in the area of medical and pharmaceutical sciences, there is still a wide knowledge gap regarding the potential application of chitosan for encapsulation of active ingredients in agriculture. Hence, the present article describes the current status of chitosan nanoparticle-based delivery systems in agriculture, and to highlight challenges that need to be overcome. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. An emerging platform for drug delivery: aerogel based systems.

    Science.gov (United States)

    Ulker, Zeynep; Erkey, Can

    2014-03-10

    Over the past few decades, advances in "aerogel science" have provoked an increasing interest for these materials in pharmaceutical sciences for drug delivery applications. Because of their high surface areas, high porosities and open pore structures which can be tuned and controlled by manipulation of synthesis conditions, nanostructured aerogels represent a promising class of materials for delivery of various drugs as well as enzymes and proteins. Along with biocompatible inorganic aerogels and biodegradable organic aerogels, more complex systems such as surface functionalized aerogels, composite aerogels and layered aerogels have also been under development and possess huge potential. Emphasis is given to the details of the aerogel synthesis and drug loading methods as well as the influence of synthesis parameters and loading methods on the adsorption and release of the drugs. Owing to their ability to increase the bioavailability of low solubility drugs, to improve both their stability and their release kinetics, there are an increasing number of research articles concerning aerogels in different drug delivery applications. This review presents an up to date overview of the advances in all kinds of aerogel based drug delivery systems which are currently under investigation. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Nanoparticle-based drug delivery systems: promising approaches against infections

    Energy Technology Data Exchange (ETDEWEB)

    Ranghar, Shweta; Sirohi, Parul [Department of Applied Mechanics, Motilal Nehru National Institute of Technology, Allahabad (India); Verma, Pritam; Agarwal, Vishnu, E-mail: vishnu_agarwal02@rediffmail.com [Department of Biotechnology, Motilal Nehru National Institute of Technology, Allahabad (India)

    2014-03-15

    Despite the fact that many new drugs and technologies have been developed to combat the infectious diseases, these have continued to be global health challenges. The use of conventional antimicrobial agents against these infections is always associated with problems such as the development of multiple drug resistance and adverse side effects. In addition, the inefficient traditional drug delivery system results in inadequate therapeutic index, low bioavailability of drugs and many other limitations. In this regard, antimicrobial nanoparticles and nanosized drug delivery carriers have emerged as potent effective agents against the infections. Nanoparticles have unique properties owing to their ultra small and controllable size such as high surface area, enhanced reactivity, and functionalizable structure. This review focused on different classes of antimicrobial nanoparticles, including metal, metal oxide and others along with their mechanism of action and their potential use against the infections. The review also focused on the development of nanoparticle systems for antimicrobial drug delivery and use of these systems for delivery of various antimicrobial agents, giving an overview about modern nanoparticle based therapeutic strategies against the infections. (author)

  17. Molecular electroporation and the transduction of oligoarginines

    Science.gov (United States)

    Cahill, Kevin

    2010-03-01

    Certain short polycations, such as TAT and polyarginine, rapidly pass through the plasma membranes of mammalian cells by an unknown mechanism called transduction as well as by endocytosis and macropinocytosis. These cell-penetrating peptides (CPPs) promise to be medically useful when fused to biologically active peptides. I offer a simple model in which one or more CPPs and the phosphatidylserines of the inner leaflet form a kind of capacitor with a voltage in excess of about 200 mV, high enough to create a molecular electropore. The model is consistent with an empirical upper limit on the cargo peptide of 40-60 amino acids and with experimental data on how the transduction of a polyarginine-fluorophore into mouse C2C12 myoblasts depends on the number of arginines in the CPP and on the CPP concentration. The model makes three testable predictions.

  18. Case report: Irreversible electroporation for locally advanced pancreatic cancer

    Directory of Open Access Journals (Sweden)

    Sonia Orcutt

    2017-01-01

    Conclusion: While there is a positive outlook for the use of irreversible electroporation for locally advanced pancreas cancer, there remain some uncertainties surrounding this therapy, which underscores the importance of future research in this area.

  19. Normal and malignant cells exhibit differential responses to calcium electroporation

    DEFF Research Database (Denmark)

    Frandsen, Stine Krog; Krüger, Mie Barthold; Mangalanathan, Uma M.

    2017-01-01

    Calcium electroporation may offer a simple general tool for anticancer therapy. Transient permeabilization of cancer cell membranes created by applying short, high-voltage pulses in tumors enables high calcium influxes that trigger cell death. In this study, we compared the relative sensitivity...... of different human tumor models and normal tissues to calcium electroporation. Plasma membrane Ca(2+)-ATPase (PMCA) protein expression was confirmed in vitro in all cancer cell lines and normal primary dermal fibroblasts studied. In all tumor types tested in vivo, calcium electroporation effectively induced...... tissue after calcium electroporation but decreased in skin tissue 4 hours after treatment to levels comparable with untreated controls, whereas calcium content endured at high levels in tumor tissue. Mechanistic experiments in vitro indicated that calcium influx was similar in fibroblasts and cancer...

  20. A Rule-Based System for Contextualized Information Delivery

    OpenAIRE

    Hu, B.; Hidders, A.J.H.; De Lignie, M.; Cimiano, P.

    2011-01-01

    When carrying out tasks, police officers need up-to-date information contextualized to their current situation to support them in decision making. The results of a previous user study with the aim of capturing the information requirements of police officers have led to the implementation of a rule-based system for contextualized information delivery. In this paper, we present the overall system and discuss how the various sources of information are modelled using ontologies. Our focus is on t...

  1. Rapid Single-Cell Electroporation for Labeling Organotypic Cultures

    Science.gov (United States)

    2010-06-01

    EMCDD (Andor), a 600x800 color camera ( MVC ), and 640x480 analog video camera (Hamamatsu), and a 1024x1024 cooled camera (Photometrics CoolSNAP). A custom...Preparation and Loading of Protein Samples for Microinjection. Cold Spring Harbor Protocols (2007). 21. Schneckenburger, H., Hendinger, A., Sailer...Electroporation Method for Mammalisan CNS Neurons in Organotypic Slice Cultures. in Electroporation and Sonoporation in Developmental Biology 169-177 ( Springe

  2. Alginate-based hybrid aerogel microparticles for mucosal drug delivery.

    Science.gov (United States)

    Gonçalves, V S S; Gurikov, P; Poejo, J; Matias, A A; Heinrich, S; Duarte, C M M; Smirnova, I

    2016-10-01

    The application of biopolymer aerogels as drug delivery systems (DDS) has gained increased interest during the last decade since these structures have large surface area and accessible pores allowing for high drug loadings. Being biocompatible, biodegradable and presenting low toxicity, polysaccharide-based aerogels are an attractive carrier to be applied in pharmaceutical industry. Moreover, some polysaccharides (e.g. alginate and chitosan) present mucoadhesive properties, an important feature for mucosal drug delivery. This feature allows to extend the contact of DDS with biological membranes, thereby increasing the absorption of drugs through the mucosa. Alginate-based hybrid aerogels in the form of microparticles (aerogel microparticles were obtained for alginate, hybrid alginate/pectin and alginate/κ-carrageenan aerogels, presenting high specific surface area (370-548m(2)g(-1)) and mucoadhesive properties. The microparticles were loaded with ketoprofen via adsorption from its solution in sc-CO2, and with quercetin via supercritical anti-solvent precipitation. Loading of ketoprofen was in the range between 17 and 22wt% whereas quercetin demonstrated loadings of 3.1-5.4wt%. Both the drugs were present in amorphous state. Loading procedure allowed the preservation of antioxidant activity of quercetin. Release of both drugs from alginate/κ-carrageenan aerogel was slightly faster compared to alginate/pectin. The results indicate that alginate-based aerogel microparticles can be viewed as promising matrices for mucosal drug delivery applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Particle-based delivery of the HIV envelope protein.

    Science.gov (United States)

    Asbach, Benedikt; Wagner, Ralf

    2017-05-01

    A major focus in HIV vaccine research is the development of suitable antigens that elicit broadly neutralizing antibody responses targeting HIV's envelope protein (Env). Delivery of Env in a repetitive manner on particle-based carriers allows higher avidity interactions and is therefore expected to efficiently engage B cells, thus leading to affinity maturation that results in superior antibody responses characterized by improved breadth, potency, and durability. This review summarizes current work that is evaluating diverse types of such particulate carriers for Env delivery. Various types of particle scaffolds are being investigated, encompassing group-specific antigen-derived virus-like particles, bacteria-derived proteins that self-assemble into symmetrical nanoparticles, as well as liposomes assembled from membrane components and recombinantly produced Env isoforms. Env-derived antigens from peptides over selected isolates to improved, stabilized next-generation designer Envs have been attached to such carriers. Immunological evaluation in animal models showed that these structures often elicit superior humoral immune responses. The findings reviewed here emphasize the potential of particle-based delivery modalities to elicit better antibody responses. Together with advances in Env antigen design, these approaches may synergistically act together on the way to obtain vaccine candidates that potentially induce protective immune responses against HIV.

  4. Delivery of RNAi-Based Oligonucleotides by Electropermeabilization

    Directory of Open Access Journals (Sweden)

    Muriel Golzio

    2013-04-01

    Full Text Available For more than a decade, understanding of RNA interference (RNAi has been a growing field of interest. The potent gene silencing ability that small oligonucleotides have offers new perspectives for cancer therapeutics. One of the present limits is that many biological barriers exist for their efficient delivery into target cells or tissues. Electropermeabilization (EP is one of the physical methods successfully used to transfer small oligonucleotides into cells or tissues. EP consists in the direct application of calibrated electric pulses to cells or tissues that transiently permeabilize the plasma membranes, allowing efficient in vitro and in vivo. cytoplasmic delivery of exogenous molecules. The present review reports on the type of therapeutic RNAi-based oligonucleotides that can be electrotransferred, the mechanism(s of their electrotransfer and the technical settings for pre-clinical purposes.

  5. Collaborative Product Design for Tasks Sorting Based on Shortest Delivery

    Directory of Open Access Journals (Sweden)

    Xuedong Liang

    2016-01-01

    Full Text Available The “people’s innovation” can promote development in enterprises in urgent need of innovative product design. Collaborative product design can be a powerful tool for enterprises wishing to improve their market competitiveness and customer satisfaction. To reduce decision costs, improve efficiency, and solve other issues, promoting “people’s innovation” can play a vital role. With this focus, this paper examines products produced through “people’s innovation.” A collaborative design task scheduling problem is presented. The design tasks are sorted based on a minimum delivery cost principle, which is determined using weighted shortest processing time (WSPT rules and the shortest delivery time. The results show that distributed collaborative innovation can result in a reasonable arrangement for collaborative design tasks.

  6. New perspectives on lipid and surfactant based drug delivery systems for oral delivery of poorly soluble drugs

    DEFF Research Database (Denmark)

    Müllertz, Anette; Ogbonna, Anayo; Ren, Shan

    2010-01-01

    The aim of this review is to highlight relevant considerations when implementing a rational strategy for the development of lipid and surfactant based drug delivery system and to discuss shortcomings and challenges to the current classification of these delivery systems. We also aim to offer...... suggestions for an improved classification system that will accommodate lipid based formulations that are not currently accommodated in the lipid formulation classification system....

  7. A theoretical analysis of the feasibility of a singularity-induced micro-electroporation system.

    Directory of Open Access Journals (Sweden)

    Gregory D Troszak

    Full Text Available Electroporation, the permeabilization of the cell membrane lipid bilayer due to a pulsed electric field, has important implications in the biotechnology, medicine, and food industries. Traditional macro and micro-electroporation devices have facing electrodes, and require significant potential differences to induce electroporation. The goal of this theoretical study is to investigate the feasibility of singularity-induced micro-electroporation; an electroporation configuration aimed at minimizing the potential differences required to induce electroporation by separating adjacent electrodes with a nanometer-scale insulator. In particular, this study aims to understand the effect of (1 insulator thickness and (2 electrode kinetics on electric field distributions in the singularity-induced micro-electroporation configuration. A non-dimensional primary current distribution model of the micro-electroporation channel shows that while increasing insulator thickness results in smaller electric field magnitudes, electroporation can still be performed with insulators thick enough to be made with microfabrication techniques. Furthermore, a secondary current distribution model of the singularity-induced micro-electroporation configuration with inert platinum electrodes and water electrolyte indicates that electrode kinetics do not inhibit charge transfer to the extent that prohibitively large potential differences are required to perform electroporation. These results indicate that singularity-induced micro-electroporation could be used to develop an electroporation system that consumes minimal power, making it suitable for remote applications such as the sterilization of water and other liquids.

  8. Multilayered materials based on biopolymers as drug delivery systems.

    Science.gov (United States)

    Vilela, Carla; Figueiredo, Ana R P; Silvestre, Armando J D; Freire, Carmen S R

    2017-02-01

    The design of efficient therapeutic delivery devices has become a tremendously active area of research with a strong contribution from the layer-by-layer (LbL) technology. The application of this simple yet firmly established technique for the design of drug reservoirs originates a multitude of multilayered systems of tailored architecture and with a high level of control of drug administration. Areas covered: This review will focus on the most recent and original research on LbL assemblies based on biopolymers including polysaccharides, polypeptides and proteins, with potential use in drug delivery. Herein, drug reservoirs consisting of multilayered planar films and capsules will be examined with emphasis on the ones benefiting from the non-cytotoxic and biocompatible nature of biopolymers, which are suitable to load, protect and release a high payload of toxic and fragile drugs. Expert opinion: The combination of biopolymers with LbL technology has undergone extensive research, still, there is a multitude of R&D opportunities for the design of smart drug delivery systems with distinct multilayered morphologies, low immunological response, non-invasive drug release devices, as well as the design of theranostic systems combining diagnostics and therapeutic features. Further developments in terms of scaling towards mass production in the pharmaceutical industry are expected in the long-term.

  9. Silk Fibroin-Based Nanoparticles for Drug Delivery

    Science.gov (United States)

    Zhao, Zheng; Li, Yi; Xie, Mao-Bin

    2015-01-01

    Silk fibroin (SF) is a protein-based biomacromolecule with excellent biocompatibility, biodegradability and low immunogenicity. The development of SF-based nanoparticles for drug delivery have received considerable attention due to high binding capacity for various drugs, controlled drug release properties and mild preparation conditions. By adjusting the particle size, the chemical structure and properties, the modified or recombinant SF-based nanoparticles can be designed to improve the therapeutic efficiency of drugs encapsulated into these nanoparticles. Therefore, they can be used to deliver small molecule drugs (e.g., anti-cancer drugs), protein and growth factor drugs, gene drugs, etc. This paper reviews recent progress on SF-based nanoparticles, including chemical structure, properties, and preparation methods. In addition, the applications of SF-based nanoparticles as carriers for therapeutic drugs are also reviewed. PMID:25749470

  10. Silk Fibroin-Based Nanoparticles for Drug Delivery

    Directory of Open Access Journals (Sweden)

    Zheng Zhao

    2015-03-01

    Full Text Available Silk fibroin (SF is a protein-based biomacromolecule with excellent biocompatibility, biodegradability and low immunogenicity. The development of SF-based nanoparticles for drug delivery have received considerable attention due to high binding capacity for various drugs, controlled drug release properties and mild preparation conditions. By adjusting the particle size, the chemical structure and properties, the modified or recombinant SF-based nanoparticles can be designed to improve the therapeutic efficiency of drugs encapsulated into these nanoparticles. Therefore, they can be used to deliver small molecule drugs (e.g., anti-cancer drugs, protein and growth factor drugs, gene drugs, etc. This paper reviews recent progress on SF-based nanoparticles, including chemical structure, properties, and preparation methods. In addition, the applications of SF-based nanoparticles as carriers for therapeutic drugs are also reviewed.

  11. Microneedles As a Delivery System for Gene Therapy

    Science.gov (United States)

    Chen, Wei; Li, Hui; Shi, De; Liu, Zhenguo; Yuan, Weien

    2016-01-01

    Gene delivery systems can be divided to two major types: vector-based (either viral vector or non-viral vector) and physical delivery technologies. Many physical carriers, such as electroporation, gene gun, ultrasound start to be proved to have the potential to enable gene therapy. A relatively new physical delivery technology for gene delivery consists of microneedles (MNs), which has been studied in many fields and for many molecule types and indications. Microneedles can penetrate the stratum corneum, which is the main barrier for drug delivery through the skin with ease of administration and without significant pain. Many different kinds of MNs, such as metal MNs, coated MNs, dissolving MNs have turned out to be promising in gene delivery. In this review, we discussed the potential as well as the challenges of utilizing MNs to deliver nucleic acids for gene therapy. We also proposed that a combination of MNs and other gene delivery approaches may lead to a better delivery system for gene therapy. PMID:27303298

  12. Microneedles as a Delivery System for Gene Therapy

    Directory of Open Access Journals (Sweden)

    Wei eChen

    2016-05-01

    Full Text Available Gene delivery systems can be divided to two major types: vector-based (either viral vector or non-viral vector and physical delivery technologies. Many physical carriers, such as electroporation, gene gun, ultrasound start to be proved to have the potential to enable gene therapy. A relatively new physical delivery technology for gene delivery consists of microneedles (MNs, which has been studied in many fields and for many molecule types and indications. Microneedles can penetrate the stratum corneum, which is the main barrier for drug delivery through the skin with ease of administration and without significant pain. Many different kinds of MNs, such as metal MNs, coated MNs, dissolving MNs have turned out to be promising in gene delivery. In this review, we discussed the potential as well as the challenges of utilizing MNs to deliver nucleic acids for gene therapy. We also proposed that a combination of MNs and other gene delivery approaches may lead to a better delivery system for gene therapy.

  13. Application of Electroporation Technique in Biofuel Processing

    Directory of Open Access Journals (Sweden)

    Yousuf Abu

    2017-01-01

    Full Text Available Biofuels production is mostly oriented with fermentation process, which requires fermentable sugar as nutrient for microbial growth. Lignocellulosic biomass (LCB represents the most attractive, low-cost feedstock for biofuel production, it is now arousing great interest. The cellulose that is embedded in the lignin matrix has an insoluble, highly-crystalline structure, so it is difficult to hydrolyze into fermentable sugar or cell protein. On the other hand, microbial lipid has been studying as substitute of plant oils or animal fat to produce biodiesel. It is still a great challenge to extract maximum lipid from microbial cells (yeast, fungi, algae investing minimum energy.Electroporation (EP of LCB results a significant increase in cell conductivity and permeability caused due to the application of an external electric field. EP is required to alter the size and structure of the biomass, to reduce the cellulose crystallinity, and increase their porosity as well as chemical composition, so that the hydrolysis of the carbohydrate fraction to monomeric sugars can be achieved rapidly and with greater yields. Furthermore, EP has a great potential to disrupt the microbial cell walls within few seconds to bring out the intracellular materials (lipid to the solution. Therefore, this study aims to describe the challenges and prospect of application of EP technique in biofuels processing.

  14. Potential for layered double hydroxides-based, innovative drug delivery systems

    National Research Council Canada - National Science Library

    Zhang, Kai; Xu, Zhi Ping; Lu, Ji; Tang, Zhi Yong; Zhao, Hui Jun; Good, David A; Wei, Ming Qian

    2014-01-01

    Layered Double Hydroxides (LDHs)-based drug delivery systems have, for many years, shown great promises for the delivery of chemical therapeutics and bioactive molecules to mammalian cells in vitro and in vivo...

  15. Normal and Malignant Cells Exhibit Differential Responses to Calcium Electroporation.

    Science.gov (United States)

    Frandsen, Stine K; Krüger, Mie B; Mangalanathan, Uma M; Tramm, Trine; Mahmood, Faisal; Novak, Ivana; Gehl, Julie

    2017-08-15

    Calcium electroporation may offer a simple general tool for anticancer therapy. Transient permeabilization of cancer cell membranes created by applying short, high-voltage pulses in tumors enables high calcium influxes that trigger cell death. In this study, we compared the relative sensitivity of different human tumor models and normal tissues to calcium electroporation. Plasma membrane Ca(2+)-ATPase (PMCA) protein expression was confirmed in vitro in all cancer cell lines and normal primary dermal fibroblasts studied. In all tumor types tested in vivo, calcium electroporation effectively induced necrosis, with a range of sensitivities observed (36%-88%) 2 days after treatment. Necrosis was induced using calcium concentrations of 100-500 mmol/L and injection volumes 20%-80% of tumor volume. Notably, only limited effects were seen in normal tissue. Calcium content increased >7-fold in tumor and skin tissue after calcium electroporation but decreased in skin tissue 4 hours after treatment to levels comparable with untreated controls, whereas calcium content endured at high levels in tumor tissue. Mechanistic experiments in vitro indicated that calcium influx was similar in fibroblasts and cancer cells. However, we observed decreased PMCA expression in cancer cells compared with fibroblasts, offering a potential explanation for the different calcium content in tumor cells versus normal tissues. Overall, our results suggest that calcium electroporation can elicit a rapid and selective necrosis of solid tumors, with limited deleterious effects on surrounding normal tissues. Cancer Res; 77(16); 4389-401. ©2017 AACR. ©2017 American Association for Cancer Research.

  16. Reasons For Preference Of Delivery In Spiritual Church-Based ...

    African Journals Online (AJOL)

    Results: Various reasons for preferring church delivery included: Spiritual protection against satanic attacks and safe delivery in 975 (36.8%) lack of funds in 629(30.5%), harsh attitude of health workers in 249 (12.1%), convenience in 212 (10.3%), faith in God and previous delivery in church 83 (4.0%) each help and good ...

  17. Simple Genome Editing of Rodent Intact Embryos by Electroporation.

    Directory of Open Access Journals (Sweden)

    Takehito Kaneko

    Full Text Available The clustered regularly interspaced short palindromic repeat (CRISPR/CRISPR-associated (Cas system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN comprised the edited targeted gene as a knockout (67% of mice and 88% of rats or knock-in (both 33%. The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.

  18. Transfer of Foreign DNA into Aquatic Animals by Electroporation

    Science.gov (United States)

    Chen, Thomas T.; Chen, Maria J.; Chiou, Tzu-Ting; Lu, J. K.

    Aquatic animals into which a foreign gene or a non-coding DNA fragment is artificially introduced and integrated in their genomes are called transgenic aquatic animals. Since 1985, a wide range of transgenic aquatic animal species have been produced mainly by microinjecting or electroporating homologous or heterologous transgenes into newly fertilized or unfertilized eggs and sometimes, sperm (for review, Chen and Powers, 1990; Hackett, 1993; Chiou et al., 2005). To produce a desired transgenic aquatic animal species, several factors should be considered. First, could the reproduction cycle of the aquatic animal species under consideration be completed in captivity? Second, a specific gene construct must be designed based on the special requirements of each study. For example, the gene construct may contain an open reading frame encoding a gene product of interest and regulatory elements that regulate the expression of the gene in a temporal, spatial and/or devel opmental manner. Third, an efficient method for delivering the transgene construct needs to be identified. Fourth, since not all instances of gene transfer are efficient, a screening method must be adopted for identifying transgenic individuals.

  19. PH responsive polypeptide based polymeric micelles for anticancer drug delivery.

    Science.gov (United States)

    Zhao, Dongping; Li, Bingqiang; Han, Jiaming; Yang, Yue; Zhang, Xinchen; Wu, Guolin

    2015-09-01

    A pH-responsive polymeric micelle based on poly(aspartamide) derivative was explored as an efficient acid-triggered anticancer drug delivery system. Poly(α,β-l-asparthydrazide) (PAHy) was prepared by aminolysis reaction of polysuccinimide with hydrazine hydrate. Poly(ethylene glycol) and aliphatic chain (C18) were conjugated onto PAHy to afford an amphiphilic copolymer with acid-liable hydrazone bonds. The structure of the resulting copolymer and its self-assembled micelles were confirmed by (1) H NMR, FTIR, DLS, and TEM. Furthermore, doxorubicin (DOX) was loaded into the polymeric micelles via the hydrophobic interaction between the C18 group and DOX molecules, and the π-π staking between the hydrazone conjugated DOX and free DOX molecules. Results showed that the DOX loaded nanoparticle (NP) was relatively stable under physiological conditions, while the DOX was quickly released in response to acidity due to the shedding of mPEG shells and dissociating of C18 segments because of the pH-cleavage of intermediate hydrazone bonds. In addition, the DOX loaded micelles presented a high cytotoxic activity against tumor cells in vitro. This pH responsive NP has appeared highly promising for the targeted intracellular delivery of hydrophobic chemotherapeutics in cancer therapy. © 2015 Wiley Periodicals, Inc.

  20. Injectable biopolymer based hydrogels for drug delivery applications.

    Science.gov (United States)

    Atta, Sadia; Khaliq, Shaista; Islam, Atif; Javeria, Irtaza; Jamil, Tahir; Athar, Muhammad Makshoof; Shafiq, Muhammad Imtiaz; Ghaffar, Abdul

    2015-09-01

    Biopolymer based pH-sensitive hydrogels were prepared using chitosan (CS) with polyethylene glycol (PEG) of different molecular weights in the presence of silane crosslinker. The incorporated components remain undissolved in different swelling media as they are connected by siloxane linkage which was confirmed by Fourier transform infrared spectroscopy. The swelling in water was enhanced by the addition of higher molecular weight PEG. The swelling behaviour of the hydrogels against pH showed high swelling in acidic and basic pH, whereas, low swelling was examined at pH 6 and 7. This characteristic pH responsive behaviour at neutral pH made them suitable for injectable controlled drug delivery. The controlled release analysis of Cefixime (CFX) (model drug) loaded CS/PEG hydrogel exhibited that the entire drug was released in 30 min in simulated gastric fluid (SGF) while in simulated intestinal fluid (SIF), 85% of drug was released in controlled manner within 80 min. This inferred that the developed hydrogels can be an attractive biomaterial for injectable drug delivery with physiological pH and other biomedical applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Recent advances in chitosan-based nanoparticulate pulmonary drug delivery

    Science.gov (United States)

    Islam, Nazrul; Ferro, Vito

    2016-07-01

    The advent of biodegradable polymer-encapsulated drug nanoparticles has made the pulmonary route of administration an exciting area of drug delivery research. Chitosan, a natural biodegradable and biocompatible polysaccharide has received enormous attention as a carrier for drug delivery. Recently, nanoparticles of chitosan (CS) and its synthetic derivatives have been investigated for the encapsulation and delivery of many drugs with improved targeting and controlled release. Herein, recent advances in the preparation and use of micro-/nanoparticles of chitosan and its derivatives for pulmonary delivery of various therapeutic agents (drugs, genes, vaccines) are reviewed. Although chitosan has wide applications in terms of formulations and routes of drug delivery, this review is focused on pulmonary delivery of drug-encapsulated nanoparticles of chitosan and its derivatives. In addition, the controversial toxicological effects of chitosan nanoparticles for lung delivery will also be discussed.

  2. Smart drug delivery injector microsystem based on pyrotechnical actuation

    Science.gov (United States)

    Puig-Vidal, Manel; Lopez, Jaime; Miribel, Pere; Samitier-Marti, Josep; Rossi, Carole; Berthold, Axel

    2003-04-01

    A smart drug delivery injector microsystem is presented based on small pyrotechnics to impulse drugs to be injected to a human being. The proposal refers to a feasibility demonstration of the technology for pharmaceutical chips. These chips would be around some cm2 in section and will be able to inject a drug into de subject skin responding to an electrical signal. The product derived from this activity will be useful for astronaut's health, being able to administrate emergency doses of products (for instance cardio-tonic or hypoallegic drugs) enough to survive an emergency situation (as it can be a heart attack during EVA). The system can also be used for easy administration of drugs needed for physiological research. The usefulness of the device in terrestrial applications has no doubt, allowing remote administration of drugs to patients whose biomedical parameters are remotely monitored. The concept proposed here is new in combining the idea of pharmaceutical chip with the ultrasonic droplet technology and the use of pyrotechnics to provide energy to the drug to be injected. The proposed Drug Injector Microsystem is based on 2 main blocks:- Micropyrotechnic system: defines the ignition part based on pyrotechnic.- Microfluidic system: defines the drug injection part. This part is also divided in different critical parts: Expansion chamber, membrane or piston, drug reservoir and a needle. Different sensors are placed on the expansion chamber of microfluidic system and on the micropyrotechnic system. A complete electronic module is implemented with a PC interface to define flexible and user friendly experiences showing the smart drug delivery injector microsystem principle.

  3. Optimization of conditions for gene delivery system based on PEI

    Directory of Open Access Journals (Sweden)

    Roya Cheraghi

    2017-01-01

    Full Text Available Objective(s: PEI based nanoparticle (NP due to dual capabilities of proton sponge and DNA binding is known as powerful tool for nucleic acid delivery to cells. However, serious cytotoxicity and complicated conditions, which govern NPs properties and its interactions with cells practically, hindered achievement to high transfection efficiency. Here, we have tried to optimize the properties of PEI/ firefly luciferase plasmid complexes and cellular condition to improve transfection efficiency. Materials and Methods: For this purpose, firefly luciferase, as a robust gene reporter, was complexed with PEI to prepare NPs with different size and charge. The physicochemical properties of nanoparticles were evaluated using agarose gel retardation and dynamic light scattering.  MCF7 and BT474 cells at different confluency were also transfected with prepared nanoparticles at various concentrations for short and long times. Results: The branched PEI can instantaneously bind to DNA and form cationic NPs. The results demonstrated the production of nanoparticles with size about 100-500 nm dependent on N/P ratio. Moreover, increase of nanoparticles concentration on the cell surface drastically improved the transfection rate, so at a concentration of 30 ng/ìl, the highest transfection efficiency was achieved. On the other side, at confluency between 40-60%, the maximum efficiency was obtained. The result demonstrated that N/P ratio of 12 could establish an optimized ratio between transfection efficiency and cytotoxicity of PEI/plasmid nanoparticles. The increase of NPs N/P ratio led to significant cytotoxicity. Conclusion: Obtained results verified the optimum conditions for PEI based gene delivery in different cell lines.

  4. Chitosan-Based Nanoparticles for Mucosal Delivery of RNAi Therapeutics

    DEFF Research Database (Denmark)

    Martirosyan, Alina; Olesen, Morten Jarlstad; Howard, Kenneth A.

    2014-01-01

    of the polysaccharide chitosan have been used to facilitate delivery of siRNA across mucosal surfaces following local administration. This chapter describes the mucosal barriers that need to be addressed in order to design an effective mucosal delivery strategy and the utilization of the mucoadhesive properties...... of chitosan. Focus is given to preparation methods and the preclinical application of chitosan nanoparticles for respiratory and oral delivery of siRNA....

  5. Nucleoside-Lipid-Based Nanocarriers for Sorafenib Delivery

    Science.gov (United States)

    Benizri, Sebastien; Ferey, Ludivine; Alies, Bruno; Mebarek, Naila; Vacher, Gaelle; Appavoo, Ananda; Staedel, Cathy; Gaudin, Karen; Barthélémy, Philippe

    2018-01-01

    Although the application of sorafenib, a small inhibitor of tyrosine protein kinases, to cancer treatments remains a worldwide option in chemotherapy, novel strategies are needed to address the low water solubility (side effects issues of this drug. In this context, the use of nanocarriers is currently investigated in order to overcome these drawbacks. In this contribution, we report a new type of sorafenib-based nanoparticles stabilized by hybrid nucleoside-lipids. The solid lipid nanoparticles (SLNs) showed negative or positive zeta potential values depending on the nucleoside-lipid charge. Transmission electron microscopy of sorafenib-loaded SLNs revealed parallelepiped nanoparticles of about 200 nm. Biological studies achieved on four different cell lines, including liver and breast cancers, revealed enhanced anticancer activities of Sorafenib-based SLNs compared to the free drug. Importantly, contrast phase microscopy images recorded after incubation of cancer cells in the presence of SLNs at high concentration in sorafenib (> 80 μM) revealed a total cancer cell death in all cases. These results highlight the potential of nucleoside-lipid-based SLNs as drug delivery systems.

  6. Low vulnerability of the right phrenic nerve to electroporation ablation

    NARCIS (Netherlands)

    van Driel, Vincent J. H. M.; Neven, KGEJ; van Wessel, Harri; Vink, Aryan; Doevendans, Pieter A. F. M.; Wittkampf, Fred H. M.

    BACKGROUND Circular electroporation ablation is a novel ablation modality for electrical pulmonary vein isolation. With a single 200-3 application, deep circular myocardial lesions can be created. However, the acute and chronic effects of this energy source on phrenic nerve (PN) function are

  7. Gene transfer into mouse prepancreatic endoderm by whole embryo electroporation.

    Science.gov (United States)

    Pierreux, Christophe E; Poll, Aurélie V; Jacquemin, Patrick; Lemaigre, Frédéric P; Rousseau, Guy G

    2005-03-10

    Understanding gene function in the developing pancreas is a major issue for pancreatic cell therapy. The in vivo analysis of gene function has essentially been performed by modulating gene expression in transgenesis. A faster and easier method is electroporation of mouse embryos. This technique, coupled with whole embryo culture, enables one to deliver genes and analyze their effects in a spatially and temporally regulated manner. We wanted to adapt the electroporation technique for gene transfer of whole e8.5 mouse embryos into the endoderm to allow expression of transgenes in the pancreas or liver. Using two platinum plate electrodes, low voltage and a precise positioning of the embryo in the electroporation cuvette we could target and express DNA constructs in the prepancreatic or prehepatic territories, identified with cell markers. We also demonstrated that this technique is a valuable tool in the study of transcriptional regulation in the developing endoderm. Targeted electroporation of whole embryos is a useful method of characterizing the gene network which controls pancreatic development.

  8. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    NARCIS (Netherlands)

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P.; de, Vicente A.

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool

  9. 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.

  10. The optimization of voltage parameter for tissue electroporation in ...

    African Journals Online (AJOL)

    USER

    2010-07-19

    Jul 19, 2010 ... Full Length Research Paper. The optimization of voltage parameter for tissue .... After incubation, 20 embryos were washed twice with sterile water and transferred to each ice-cold 0.4 cm ... were electroporated different somatic embryos of coffee, found the highest transient GUS gene expression at the.

  11. Electroporation of cells in microfluidic devices: a review

    NARCIS (Netherlands)

    Fox, M.B.; Esveld, D.C.; Valero, A.; Luttge, R.; Mastwijk, H.C.; Bartels, P.V.; Berg, van den A.B.A.; Boom, R.M.

    2006-01-01

    In recent years, several publications on microfluidic devices have focused on the process of electroporation, which results in the poration of the biological cell membrane. The devices involved are designed for cell analysis, transfection or pasteurization. The high electric field strengths needed

  12. Electroporation of cells in microfluidic devices: a review

    NARCIS (Netherlands)

    Fox, M.B.; Fox, M.B.; Esveld, D.C.; Valero, Ana; Lüttge, Regina; Mastwijk, H.C.; Bartels, P.V.; van den Berg, Albert; Boom, R.M.; Boom, R.M.

    In recent years, several publications on microfluidic devices have focused on the process of electroporation, which results in the poration of the biological cell membrane. The devices involved are designed for cell analysis, transfection or pasteurization. The high electric field strengths needed

  13. Microneedle-based drug delivery: studies on delivery parameters and biocompatibility.

    Science.gov (United States)

    Wu, Yan; Qiu, Yuqin; Zhang, Suohui; Qin, Guangjiong; Gao, Yunhua

    2008-10-01

    There is a significant interest in the application of microneedles in intradermal drug delivery systems. Previous studies have demonstrated that skin permeation of drugs can be increased by orders of magnitude with microneedle insertion. In this study, emphasis is placed on the development of low cost, painless intradermal microneedle systems that can enhance the percutaneous drug permeation. Microneedles of octagonal pyramidal shape with the length of 150 mum were employed, and the capabilities of skin permeation enhancement under different delivery conditions were examined. The delivery parameters taken into account included the insertion time and the area of insertion. It was found that when solid microneedle arrays of 150 mum in length were pierced into human dermatomed skin for 5 to 60 s, microconduits with the depth of 50 to 80 mum were created to facilitate the percutaneous permeation of drugs. In percutaneous tests, it was demonstrated that the permeability coefficient of calcein (MW = 622.55) was significantly increased by 10(4) to 10(5) times compared to that on intact skin. In terms of biocompatibility, biological evaluation indicated a broad spectrum of safety for the microneedle system. These results suggest that the octagonal pyramidal microneedles can be an effective tool in developing novel intradermal drug delivery system.

  14. The relationship between facility-based delivery and infant ...

    African Journals Online (AJOL)

    These results suggest the benefits of facility delivery transcend the immediate survival benefit for mothers and their babies, and may serve as a gateway to further interactions with the healthcare system. (Afr. J Reprod Health 2016; 20[2]: 27-33). Keywords: institutional delivery; skilled birth attendance; infant immunizations; ...

  15. Gene therapy by electroporation for the treatment of chronic renal failure in companion animals

    Directory of Open Access Journals (Sweden)

    Pope Melissa A

    2009-01-01

    Full Text Available Abstract Background Growth hormone-releasing hormone (GHRH plasmid-based therapy for the treatment of chronic renal failure and its complications was examined. Companion dogs (13.1 ± 0.8 years, 29.4 ± 5.01 kg and cats (13.2 ± 0.9 years, 8.5 ± 0.37 kg received a single 0.4 mg or 0.1 mg species-specific plasmid injection, respectively, intramuscularly followed by electroporation, and analyzed up to 75 days post-treatment; controls underwent electroporation without plasmid administration. Results Plasmid-treated animals showed an increase in body weight (dogs 22.5% and cats 3.2% compared to control animals, and displayed improved quality of life parameters including significant increases in appetite, activity, mentation and exercise tolerance levels. Insulin-like growth factor I (IGF-I, the downstream effector of GHRH levels were increased in the plasmid treated animals. Hematological parameters were also significantly improved. Protein metabolism changes were observed suggesting a shift from a catabolic to an anabolic state in the treated animals. Blood urea nitrogen and creatinine did not show any significant changes suggesting maintenance of kidney function whereas the control animal's renal function deteriorated. Treated animals survived longer than control animals with 70% of dogs and 80% of cats surviving until study day 75. Only 17% and 40% of the control dogs and cats, respectively, survived to day 75. Conclusion Improved quality of life, survival and general well-being indicate that further investigation is warranted, and show the potential of a plasmid-based therapy by electroporation in preventing and managing complications of renal insufficiency.

  16. Polymer-based nanoparticles for oral insulin delivery: Revisited approaches.

    Science.gov (United States)

    Fonte, Pedro; Araújo, Francisca; Silva, Cátia; Pereira, Carla; Reis, Salette; Santos, Hélder A; Sarmento, Bruno

    2015-11-01

    Diabetes mellitus is a high prevalence and one of the most severe and lethal diseases in the world. Insulin is commonly used to treat diabetes in order to give patients a better life condition. However, due to bioavailability problems, the most common route of insulin administration is the subcutaneous route, which may present patients compliance problems to treatment. The oral administration is thus considered the most convenient alternative to deliver insulin, but it faces important challenges. The low stability of insulin in the gastrointestinal tract and low intestinal permeation, are problems to overcome. Therefore, the encapsulation of insulin into polymer-based nanoparticles is presented as a good strategy to improve insulin oral bioavailability. In the last years, different strategies and polymers have been used to encapsulate insulin and deliver it orally. Polymers with distinct properties from natural or synthetic sources have been used to achieve this aim, and among them may be found chitosan, dextran, alginate, poly(γ-glutamic acid), hyaluronic acid, poly(lactic acid), poly(lactide-co-glycolic acid), polycaprolactone (PCL), acrylic polymers and polyallylamine. Promising studies have been developed and positive results were obtained, but there is not a polymeric-based nanoparticle system to deliver insulin orally available in the market yet. There is also a lack of long term toxicity studies about the safety of the developed carriers. Thus, the aims of this review are first to provide a deep understanding on the oral delivery of insulin and the possible routes for its uptake, and then to overview the evolution of this field in the last years of research of insulin-loaded polymer-based nanoparticles in the academic and industrial fields. Toxicity concerns of the discussed nanocarriers are also addressed. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Nanomaterial-based drug delivery carriers for cancer therapy

    CERN Document Server

    Feng, Tao

    2017-01-01

    This brief summarizes different types of organic and inorganic nanomaterials for drug delivery in cancer therapy. It highlights that precisely designed nanomaterials will be the next-generation therapeutic agents for cancer treatment.

  18. A rapid and efficient electroporation method for transformation of Halomonas sp. O-1.

    Science.gov (United States)

    Harris, Joshua R; Lundgren, Benjamin R; Grzeskowiak, Brian R; Mizuno, Kouhei; Nomura, Christopher T

    2016-10-01

    Halomonas sp. O-1 is a halophilic bacterium with a high potential for industrial application due to its natural ability to produce polyhydroxyalkanoates (PHAs) using seawater-based media. However, a major barrier preventing industrial scale implementation of this organism is a lack of molecular methodologies capable of readily transforming members of the Halomonas genus. Currently, the only reliable method used for introducing DNA into Halomonas spp. is bacterial conjugation, a somewhat tedious and time-consuming technique compared to electroporation-based methodologies. Here we describe a rapid and reproducible method for the electroporation of Halomonas sp. O-1 with plasmid DNA. Electrocompetent cells were generated by growing Halomonas sp. O-1 in a yeast extract-tryptone medium with a final salinity of 3.5%, pH of 7.5, followed by several washes using 300mM sucrose. Results show that plasmids containing chloramphenicol (Cm(R)) and gentamicin (Gm(R)) resistance cassettes are suitable antibiotic selection markers for transformation and yields of 10(4) transformants per μg of DNA were obtained. This method is simple to perform and the materials used are readily available in most research labs. Additionally, this plasmid-based transformation procedure has the potential to be adapted for a number of applications including the creation of recombinant stains and the generation of deletion mutants of Halomonas spp. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Enhanced Efficacy of a Codon-Optimized DNA Vaccine Encoding the Glycoprotein Precursor Gene of Lassa Virus in a Guinea Pig Disease Model When Delivered by Dermal Electroporation

    Directory of Open Access Journals (Sweden)

    Niranjan Y. Sardesai

    2013-07-01

    Full Text Available Lassa virus (LASV causes a severe, often fatal, hemorrhagic fever endemic to West Africa. Presently, there are no FDA-licensed medical countermeasures for this disease. In a pilot study, we constructed a DNA vaccine (pLASV-GPC that expressed the LASV glycoprotein precursor gene (GPC. This plasmid was used to vaccinate guinea pigs (GPs using intramuscular electroporation as the delivery platform. Vaccinated GPs were protected from lethal infection (5/6 with LASV compared to the controls. However, vaccinated GPs experienced transient viremia after challenge, although lower than the mock-vaccinated controls. In a follow-on study, we developed a new device that allowed for both the vaccine and electroporation pulse to be delivered to the dermis. We also codon-optimized the GPC sequence of the vaccine to enhance expression in GPs. Together, these innovations resulted in enhanced efficacy of the vaccine. Unlike the pilot study where neutralizing titers were not detected until after virus challenge, modest neutralizing titers were detected in guinea pigs before challenge, with escalating titers detected after challenge. The vaccinated GPs were never ill and were not viremic at any timepoint. The combination of the codon-optimized vaccine and dermal electroporation delivery is a worthy candidate for further development.

  20. Genomic DNA extraction from cells by electroporation on an integrated microfluidic platform.

    Science.gov (United States)

    Geng, Tao; Bao, Ning; Sriranganathanw, Nammalwar; Li, Liwu; Lu, Chang

    2012-11-06

    The vast majority of genetic analysis of cells involves chemical lysis for release of DNA molecules. However, chemical reagents required in the lysis interfere with downstream molecular biology and often require removal after the step. Electrical lysis based on irreversible electroporation is a promising technique to prepare samples for genetic analysis due to its purely physical nature, fast speed, and simple operation. However, there has been no experimental confirmation on whether electrical lysis extracts genomic DNA from cells in a reproducible and efficient fashion in comparison to chemical lysis, especially for eukaryotic cells that have most of the DNA enclosed in the nucleus. In this work, we construct an integrated microfluidic chip that physically traps a low number of cells, lyses the cells using electrical pulses rapidly, then purifies and concentrates genomic DNA. We demonstrate that electrical lysis offers high efficiency for DNA extraction from both eukaryotic cells (up to ∼36% for Chinese hamster ovary cells) and bacterial cells (up to ∼45% for Salmonella typhimurium) that is comparable to the widely used chemical lysis. The DNA extraction efficiency has dependence on both the electric parameters and relative amount of beads used for DNA adsorption. We envision that electroporation-based DNA extraction will find use in ultrasensitive assays that benefit from minimal dilution and simple procedures.

  1. Reusable titanium nitride plasmonic microstructures for intracellular delivery (Conference Presentation)

    Science.gov (United States)

    Raun, Alexander J.; Saklayen, Nabiha; Zgrabik, Christine M.; Vulis, Daryl I.; Madrid, Marinna; Shen, Weilu; Hu, Evelyn L.; Mazur, Eric

    2017-03-01

    Efficient drug and biomolecular delivery into cells is an important area of biomedical research. Intracellular delivery relies on porating cell membranes to allow exterior molecules to enter the cell efficiently and viably. Various methods, including optoporation, electroporation, and viral techniques, can deliver molecules to cells, but come with significant drawbacks such as low efficiency, low throughput, and low viability. We present a new laser-based delivery method that uses laser pulses to excite plasmonic, Titanium Nitride (TiN) microstructures for cell poration and offers high efficiency, throughput, and viability. TiN is a promising plasmonic material for laser-based delivery methods due to its high levels of hardness and thermal stability. We fabricate these microstructures by sputtering thin films of TiN on patterned sapphire substrates. We then optimize plasmonic enhancement and stability by investigating different fabrication conditions. We deliver dye molecules, siRNA, and microspheres to cells to quantify poration efficiency and viability by using flow cytometry and by imaging the target cells at defined time intervals post laser irradiation. Additionally, we study temperature effects via simulations and experiments, as well as oxidation of the TiN films over time. We also use scanning electron microscopy (SEM) techniques to study microstructure damage and cell adhesion. Overall, TiN presents a promising opportunity for use as a reusable material in future biomedical devices for intracellular biomolecular delivery and regenerative medicine.

  2. Evaluating Electroporation and Lipofectamine Approaches for Transient and Stable Transgene Expressions in Human Fibroblasts and Embryonic Stem Cells.

    Science.gov (United States)

    Sharifi Tabar, Mehdi; Hesaraki, Mahdi; Esfandiari, Fereshteh; Sahraneshin Samani, Fazel; Vakilian, Haghighat; Baharvand, Hossein

    2015-01-01

    Genetic modification of human embryonic stem cells (hESCs) is critical for their extensive use as a fundamental tool for cell therapy and basic research. Despite the fact that various methods such as lipofection and electroporation have been applied to transfer the gene of interest (GOI) into the target cell line, however, there are few re- ports that compare all parameters, which influence transfection efficiency. In this study, we examine all parameters that affect the efficiency of electroporation and lipofection for transient and long-term gene expression in three different cell lines to introduce the best method and determinant factor. In this experimental study, both electroporation and lipofection approaches were employed for genetic modification. pCAG-EGFP was applied for tran- sient expression of green fluorescent protein in two genetically different hESC lines, Roy- an H5 (XX) and Royan H6 (XY), as well as human foreskin fibroblasts (hFF). For long-term EGFP expression VASA and OLIG2 promoters (germ cell and motoneuron specific genes, respectively), were isolated and subsequently cloned into a pBluMAR5 plasmid backbone to drive EGFP expression. Flow cytometry analysis was performed two days after trans- fection to determine transient expression efficiency. Differentiation of drug resistant hESC colonies toward primordial germ cells (PGCs) was conducted to confirm stable integration of the transgene. Transient and stable expression suggested a variable potential for different cell lines against transfection. Analysis of parameters that influenced gene transformation ef- ficiency revealed that the vector concentrations from 20-60 μg and the density of the sub- jected cells (5×10(5)and 1×10(6)cells) were not as effective as the genetic background and voltage rate. The present data indicated that in contrast to the circular form, the linearized vector generated more distinctive drug resistant colonies. Electroporation was an efficient tool for genetic

  3. Achieving magneto-elasto-electroporation and cell transport using core-shell magnetoelectric nanoparticles (Conference Presentation)

    Science.gov (United States)

    Betal, Soutik; Dutta, Moumita; Shrestha, Binita; Saha, Amit; Tang, Liang; Ramasubramanian, Ananad K.; Bhalla, Amar S.; Guo, Ruyan

    2016-09-01

    Magneto-Elasto-Electroporation (MEEP) is a magnetically controlled acoustic-electroporation observed while core-shell Magneto-electric nanoparticles interact with Biological Cells. The surface polarity change of the piezoelectric coating (BaTiO3) due to absorption of pressure created due magneto-striction of core (CoFe2O4) in AC magnetic field results in electric field (Uext) change at the external vicinity of the cell membrane when nanoparticles are nearby. This results in transmembrane Voltage (Um) change which is basically the difference in Cell's internal potential (Uint) and external potential. The nonlinear permeability change of cell membrane due to change in Um opens the nano-pores on the membrane. The magnetic moment of the nanoparticles further helps in penetration of the Magneto-electric nanoparticles inside the cell through these magneto-electrically controlled newly opened nano-pores on cell's membrane. MEEP is analyzed through in-vitro analysis and Mathematical equations are formulated for numerically expressing its fundamental effect. TEM imaging, XRD analysis, Zeta-potentiometer measurement and AFM imaging are confirming the coating of the piezoelectric layer on Magneto-stricitve nanoparticles, Acoustic measurements confirms the photo-acoustic and magneto-acoustic property of CoFe2O4 nanoparticles and Fluorescence microscopy as well as Confocal microscopy are confirming the penetration of particle inside the Human Epithelial cells (HEP2). Further on application of repulsive magnetic field, nanoparticles are observed to transport a group of cells in controlled boundary conditions in microfluidic chamber. Hence these nanoparticles can be used for accurate and efficient drug delivery as well as cell transport applications

  4. IGF-I gene transfer by electroporation promotes regeneration in a muscle injury model.

    Science.gov (United States)

    Takahashi, T; Ishida, K; Itoh, K; Konishi, Y; Yagyu, K-I; Tominaga, A; Miyazaki, J-I; Yamamoto, H

    2003-04-01

    The goal of this study was to determine whether insulin-like growth factor-I (IGF-I) gene delivery by electroporation promotes repair after muscle injury. An injury-repair model was created using mice in which a hamstring muscle was cut and sutured. A total of 50 microg of IGF-I DNA or green fluorescent protein (GFP) DNA (both in pCAGGS) was injected into the lesion and introduced into muscle cells by electrostimulation using an electric pulse generator. The number of regenerating muscle fibers in the IGF-I DNA group was significantly more than that in the GFP DNA group at 2 weeks after injection. The diameter of regenerating muscle fibers from the IGF-I DNA group was larger than that of the GFP DNA group at 4 weeks after injection. There was no significant difference in the serum IGF-I concentration between the IGF-I DNA group and the GFP DNA group at 1, 2, and 4 weeks after injection. However, muscle IGF-I concentration in the IGF-I DNA injection group was significantly greater than that in the GFP DNA injection group at 2 weeks after injection. These results demonstrated that the effects of enhanced IGF-I production were local and limited to the injected area. The ratio (injected/uninjected; intact) of the amplitude of compound muscle action potentials (CMAP) in the IGF-I DNA injection group was greater than that in the GFP DNA injection group at 4 weeks after injection and of the control group. In conclusion, IGF-I gene transfer by electroporation proved to be a simple, safe, inexpensive, and effective method to promote the regeneration of injured muscles in our injury model.

  5. Selective gene expression by postnatal electroporation during olfactory interneuron neurogenesis.

    Directory of Open Access Journals (Sweden)

    Alexander T Chesler

    Full Text Available Neurogenesis persists in the olfactory system throughout life. The mechanisms of how new neurons are generated, how they integrate into circuits, and their role in coding remain mysteries. Here we report a technique that will greatly facilitate research into these questions. We found that electroporation can be used to robustly and selectively label progenitors in the Subventicular Zone. The approach was performed postnatally, without surgery, and with near 100% success rates. Labeling was found in all classes of interneurons in the olfactory bulb, persisted to adulthood and had no adverse effects. The broad utility of electroporation was demonstrated by encoding a calcium sensor and markers of intracellular organelles. The approach was found to be effective in wildtype and transgenic mice as well as rats. Given its versatility, robustness, and both time and cost effectiveness, this method offers a powerful new way to use genetic manipulation to understand adult neurogenesis.

  6. Modern medicine has a new technology: therapeutic electroporation

    Directory of Open Access Journals (Sweden)

    Lucia E. Ionescu

    2016-12-01

    Full Text Available Electroporation is considered a new start-up in the treatment of various tumors; currently, researches are being conducted in order to develop this technology with medical applications. The technique consists in the significant increase in the electrical conductivity and permeability of the plasma membrane of cells resulting from the application of an external electric field. It is routinely used in molecular biology to transform bacteria, yeast, protoplasts and is performed using the electroporators. Currently, the process seems to be a real solution that enables a targeted drug to act with maximum efficiency on cells and tissues requiring treatment, resulting in obtaining a good therapeutic effect without major side effects. Therefore, pharmaceutical companies are trying to demonstrate through preclinical studies the potential efficacy of this technology, succeeding in recent years to achieve important steps in this direction.

  7. Vaccine Delivery Methods into the Future

    Directory of Open Access Journals (Sweden)

    Vasso Apostolopoulos

    2016-03-01

    Full Text Available Several modes of vaccine delivery have been developed in the last 25 years, which induce strong immune responses in pre-clinical models and in human clinical trials. Some modes of delivery include, adjuvants (aluminum hydroxide, Ribi formulation, QS21, liposomes, nanoparticles, virus like particles, immunostimulatory complexes (ISCOMs, dendrimers, viral vectors, DNA delivery via gene gun, electroporation or Biojector 2000, cell penetrating peptides, dendritic cell receptor targeting, toll-like receptors, chemokine receptors and bacterial toxins. There is an enormous amount of information and vaccine delivery methods available for guiding vaccine and immunotherapeutics development against diseases.

  8. Electroporation Enhances the Metabolic Activity of Lactobacillus plantarum 564

    Directory of Open Access Journals (Sweden)

    Branko Bugarski

    2013-01-01

    Full Text Available The exposure of bacterial cells to pulsed electric fields (PEF leads to the reversible formation of pores in the cell membrane if an applied energy is below the critical level. Therefore, the effect of electric field pulses with amplitudes below 14 kV/cm and the applied energy up to 12.2 J/cm3 on the growth of Lactobacillus plantarum 564 cells was investigated. After PEF treatments, the growth of lactobacilli in De Man-Rogosa-Sharpe broth at 37 °C was monitored by isothermal calorimetry, absorbance and plate counts. All the applied treatments resulted in a higher growth rate of PEF-treated cells during early and mid-log phase, especially bacterial samples treated with lower field intensities (1.3–5.5 J/cm3. The transport of ions and molecules through the cell membrane (which facilitates the growth of electroporated lactobacilli was particularly evident in the mid-exponential growth phase, where the doubling time was reduced more than 3 times after the exposure to electric pulses of 5.5 J/cm3. The heat production rate during the growth of electroporated cells was also higher, indicating the enhanced metabolic activity of PEF-treated cells. Moreover, the electroporated cells had a better acidification ability than the untreated ones. It can be summarized that the applied PEF treatments with an energy input of below 12 J/cm3 potentially induce reversible electroporation of the cell membrane, which has a positive impact on the growth and metabolic activity of the cells of lactobacilli.

  9. Irreversible electroporation in primary and metastatic hepatic malignancies

    OpenAIRE

    Lyu, Tianchu; Wang, Xifu; Su, Zhanliang; Shangguan, Junjie; Sun, Chong; Figini, Matteo; Wang, Jian; Yaghmai, Vahid; LARSON, ANDREW C.; Zhang, Zhuoli

    2017-01-01

    Abstract Background: Liver cancer makes up a huge percentage of cancer mortality worldwide. Irreversible electroporation (IRE) is a relatively new minimally invasive nonthermal ablation technique for tumors that applies short pulses of high frequency electrical energy to irreversibly destabilize cell membrane to induce tumor cell apoptosis. Methods: This review aims to investigate the studies regarding the use of IRE treatment in liver tumors and metastases to liver. We searched PubMed for al...

  10. Avoiding neuromuscular stimulation in liver irreversible electroporation using radiofrequency electric fields

    Science.gov (United States)

    Castellví, Quim; Mercadal, Borja; Moll, Xavier; Fondevila, Dolors; Andaluz, Anna; Ivorra, Antoni

    2018-02-01

    Electroporation-based treatments typically consist of the application of high-voltage dc pulses. As an undesired side effect, these dc pulses cause electrical stimulation of excitable tissues such as motor nerves. The present in vivo study explores the use of bursts of sinusoidal voltage in a frequency range from 50 kHz to 2 MHz, to induce irreversible electroporation (IRE) whilst avoiding neuromuscular stimulation. A series of 100 dc pulses or sinusoidal bursts, both with an individual duration of 100 µs, were delivered to rabbit liver through thin needles in a monopolar electrode configuration, and thoracic movements were recorded with an accelerometer. Tissue samples were harvested three hours after treatment and later post-processed to determine the dimensions of the IRE lesions. Thermal damage due to Joule heating was ruled out via computer simulations. Sinusoidal bursts with a frequency equal to or above 100 kHz did not cause thoracic movements and induced lesions equivalent to those obtained with conventional dc pulses when the applied voltage amplitude was sufficiently high. IRE efficacy dropped with increasing frequency. For 100 kHz bursts, it was estimated that the electric field threshold for IRE is about 1.4 kV cm‑1 whereas that of dc pulses is about 0.5 kV cm‑1.

  11. Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes.

    Science.gov (United States)

    Chen, Sean; Lee, Benjamin; Lee, Angus Yiu-Fai; Modzelewski, Andrew J; He, Lin

    2016-07-08

    The CRISPR/Cas9 system has been employed to efficiently edit the genomes of diverse model organisms. CRISPR-mediated mouse genome editing is typically accomplished by microinjection of Cas9 DNA/RNA and single guide RNA (sgRNA) into zygotes to generate modified animals in one step. However, microinjection is a technically demanding, labor-intensive, and costly procedure with poor embryo viability. Here, we describe a simple and economic electroporation-based strategy to deliver Cas9/sgRNA ribonucleoproteins into mouse zygotes with 100% efficiency for in vivo genome editing. Our methodology, designated as CRISPR RNP Electroporation of Zygotes (CRISPR-EZ), enables highly efficient and high-throughput genome editing in vivo, with a significant improvement in embryo viability compared with microinjection. Using CRISPR-EZ, we generated a variety of editing schemes in mouse embryos, including indel (insertion/deletion) mutations, point mutations, large deletions, and small insertions. In a proof-of-principle experiment, we used CRISPR-EZ to target the tyrosinase (Tyr) gene, achieving 88% bi-allelic editing and 42% homology-directed repair-mediated precise sequence modification in live mice. Taken together, CRISPR-EZ is simple, economic, high throughput, and highly efficient with the potential to replace microinjection for in vivo genome editing in mice and possibly in other mammals. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Microfluidic-Based Synthesis of Hydrogel Particles for Cell Microencapsulation and Cell-Based Drug Delivery

    Directory of Open Access Journals (Sweden)

    Jiandi Wan

    2012-04-01

    Full Text Available Encapsulation of cells in hydrogel particles has been demonstrated as an effective approach to deliver therapeutic agents. The properties of hydrogel particles, such as the chemical composition, size, porosity, and number of cells per particle, affect cellular functions and consequently play important roles for the cell-based drug delivery. Microfluidics has shown unparalleled advantages for the synthesis of polymer particles and been utilized to produce hydrogel particles with a well-defined size, shape and morphology. Most importantly, during the encapsulation process, microfluidics can control the number of cells per particle and the overall encapsulation efficiency. Therefore, microfluidics is becoming the powerful approach for cell microencapsulation and construction of cell-based drug delivery systems. In this article, I summarize and discuss microfluidic approaches that have been developed recently for the synthesis of hydrogel particles and encapsulation of cells. I will start by classifying different types of hydrogel material, including natural biopolymers and synthetic polymers that are used for cell encapsulation, and then focus on the current status and challenges of microfluidic-based approaches. Finally, applications of cell-containing hydrogel particles for cell-based drug delivery, particularly for cancer therapy, are discussed.

  13. Irreversible Electroporation for Focal Ablation at the Porta Hepatis

    Energy Technology Data Exchange (ETDEWEB)

    Kasivisvanathan, Veeru, E-mail: vk103@ic.ac.uk [Imperial College London, Department of Radiology (United Kingdom); Thapar, Ankur, E-mail: a.thapar09@imperial.ac.uk; Oskrochi, Youssof, E-mail: Youssof.Oskrochi09@imperial.ac.uk [Imperial College London, Department of Surgery and Cancer (United Kingdom); Picard, John, E-mail: John.picard@imperial.nhs.uk [Imperial College Healthcare NHS Trust, Department of Anaesthesia (United Kingdom); Leen, Edward L. S., E-mail: Edward.leen@imperial.ac.uk [Imperial College London, Department of Radiology (United Kingdom)

    2012-12-15

    Patients with chemotherapy-refractory liver metastases who are not candidates for surgery may be treated with focal ablation techniques with established survival benefits. Irreversible electroporation is the newest of these and has the putative advantages of a nonthermal action, preventing damage to adjacent biliary structures and bowel. This report describes the use of irreversible electroporation in a 61-year-old man with a solitary chemoresistant liver metastasis unsuitable for radiofrequency ablation as a result of its proximity to the porta hepatis. At 3 months, tumor size was decreased on computed tomography from 28 Multiplication-Sign 19 to 20 Multiplication-Sign 17 mm, representing stable disease according to the response evaluation criteria in solid tumors. This corresponded to a decrease in tumor volume size from 5.25 to 3.16 cm{sup 3}. There were no early or late complications. Chemoresistant liver metastases in the proximity of the porta hepatis that are considered to be too high a risk for conventional surgery or thermal ablation may be considered for treatment by the novel ablation technique of irreversible electroporation.

  14. Development of a Gastroretentive Drug Delivery System based on ...

    African Journals Online (AJOL)

    Erah

    Purpose: The aim of this work was to synthesize superporous hydrogels of rosiglitazone using chitosan and to study its swelling behaviour for application as a gastroretentive drug delivery system. Methods: Chitosan superporous hydrogels were synthesized using glyoxal as a crosslinking agent by gas blowing method.

  15. Development of a Gastroretentive Drug Delivery System based on ...

    African Journals Online (AJOL)

    Erah

    gastroretentive drug delivery system in view of their swelling characteristics in acidic pH. Keywords: ... bioavailability of basic drugs that have poor solubility at ... polymeric materials. The water uptake of such hydrogels depends on environmental conditions (pH, ionic strength, temperature, and electrical or magnetic field).

  16. Based Mucoadhesive Gel for Nasal Delivery of Rizatriptan Benzoate

    African Journals Online (AJOL)

    Purpose: To develop a safe mucoadhesive gel for nasal drug delivery using okra polysaccharide. Methods: Rizatriptan benzoate nasal gel was developed using natural polysaccharide obtained from the Abelmoschus esculentus. The gel formulation was characterized for pH, viscosity, mucoadhesion, and in vitro ...

  17. Microprocessor Based Temperature Control of Liquid Delivery with Flow Disturbances.

    Science.gov (United States)

    Kaya, Azmi

    1982-01-01

    Discusses analytical design and experimental verification of a PID control value for a temperature controlled liquid delivery system, demonstrating that the analytical design techniques can be experimentally verified by using digital controls as a tool. Digital control instrumentation and implementation are also demonstrated and documented for…

  18. Amorphous Calcium Carbonate Based-Microparticles for Peptide Pulmonary Delivery.

    Science.gov (United States)

    Tewes, Frederic; Gobbo, Oliviero L; Ehrhardt, Carsten; Healy, Anne Marie

    2016-01-20

    Amorphous calcium carbonate (ACC) is known to interact with proteins, for example, in biogenic ACC, to form stable amorphous phases. The control of amorphous/crystalline and inorganic/organic ratios in inhalable calcium carbonate microparticles may enable particle properties to be adapted to suit the requirements of dry powders for pulmonary delivery by oral inhalation. For example, an amorphous phase can immobilize and stabilize polypeptides in their native structure and amorphous and crystalline phases have different mechanical properties. Therefore, inhalable composite microparticles made of inorganic (i.e., calcium carbonate and calcium formate) and organic (i.e., hyaluronan (HA)) amorphous and crystalline phases were investigated for peptide and protein pulmonary aerosol delivery. The crystalline/amorphous ratio and polymorphic form of the inorganic component was altered by changing the microparticle drying rate and by changing the ammonium carbonate and HA initial concentration. The bioactivity of the model peptide, salmon calcitonin (sCT), coprocessed with alpha-1-antitrypsin (AAT), a model protein with peptidase inhibitor activity, was maintained during processing and the microparticles had excellent aerodynamic properties, making them suitable for pulmonary aerosol delivery. The bioavailability of sCT after aerosol delivery as sCT and AAT-loaded composite microparticles to rats was 4-times higher than that of sCT solution.

  19. Benchmark – based review as a strategy for microfinance delivery ...

    African Journals Online (AJOL)

    Microfinance is one of the development tools for poverty reduction. The traditional supply-led subsidized credit delivery has led to increase in credit disbursements. However, there is shortage of model benchmark and indicators for evaluating and comparing performance of microfinance schemes. This study reviewed the ...

  20. Hidden Costs of Hospital Based Delivery from Two Tertiary Hospitals in Western Nepal.

    Directory of Open Access Journals (Sweden)

    Jeevan Acharya

    Full Text Available Hospital based delivery has been an expensive experience for poor households because of hidden costs which are usually unaccounted in hospital costs. The main aim of this study was to estimate the hidden costs of hospital based delivery and determine the factors associated with the hidden costs.A hospital based cross-sectional study was conducted among 384 post-partum mothers with their husbands/house heads during the discharge time in Manipal Teaching Hospital and Western Regional Hospital, Pokhara, Nepal. A face to face interview with each respondent was conducted using a structured questionnaire. Hidden costs were calculated based on the price rate of the market during the time of the study.The total hidden costs for normal delivery and C-section delivery were 243.4 USD (US Dollar and 321.6 USD respectively. Of the total maternity care expenditures; higher mean expenditures were found for food & drinking (53.07%, clothes (9.8% and transport (7.3%. For postpartum women with their husband or house head, the total mean opportunity cost of "days of work loss" were 84.1 USD and 81.9 USD for normal delivery and C-section respectively. Factors such as literate mother (p = 0.007, employed house head (p = 0.011, monthly family income more than 25,000 NRs (Nepalese Rupees (p = 0.014, private hospital as a place of delivery (p = 0.0001, C-section as a mode of delivery (p = 0.0001, longer duration (>5days of stay in hospital (p = 0.0001, longer distance (>15km from house to hospital (p = 0.0001 and longer travel time (>240 minutes from house to hospital (p = 0.007 showed a significant association with the higher hidden costs (>25000 NRs.Experiences of hidden costs on hospital based delivery and opportunity costs of days of work loss were found high. Several socio-demographic factors, delivery related factors (place and mode of delivery, length of stay, distance from hospital and travel time were associated with hidden costs. Hidden costs can be a

  1. Hidden Costs of Hospital Based Delivery from Two Tertiary Hospitals in Western Nepal.

    Science.gov (United States)

    Acharya, Jeevan; Kaehler, Nils; Marahatta, Sujan Babu; Mishra, Shiva Raj; Subedi, Sudarshan; Adhikari, Bipin

    2016-01-01

    Hospital based delivery has been an expensive experience for poor households because of hidden costs which are usually unaccounted in hospital costs. The main aim of this study was to estimate the hidden costs of hospital based delivery and determine the factors associated with the hidden costs. A hospital based cross-sectional study was conducted among 384 post-partum mothers with their husbands/house heads during the discharge time in Manipal Teaching Hospital and Western Regional Hospital, Pokhara, Nepal. A face to face interview with each respondent was conducted using a structured questionnaire. Hidden costs were calculated based on the price rate of the market during the time of the study. The total hidden costs for normal delivery and C-section delivery were 243.4 USD (US Dollar) and 321.6 USD respectively. Of the total maternity care expenditures; higher mean expenditures were found for food & drinking (53.07%), clothes (9.8%) and transport (7.3%). For postpartum women with their husband or house head, the total mean opportunity cost of "days of work loss" were 84.1 USD and 81.9 USD for normal delivery and C-section respectively. Factors such as literate mother (p = 0.007), employed house head (p = 0.011), monthly family income more than 25,000 NRs (Nepalese Rupees) (p = 0.014), private hospital as a place of delivery (p = 0.0001), C-section as a mode of delivery (p = 0.0001), longer duration (>5days) of stay in hospital (p = 0.0001), longer distance (>15km) from house to hospital (p = 0.0001) and longer travel time (>240 minutes) from house to hospital (p = 0.007) showed a significant association with the higher hidden costs (>25000 NRs). Experiences of hidden costs on hospital based delivery and opportunity costs of days of work loss were found high. Several socio-demographic factors, delivery related factors (place and mode of delivery, length of stay, distance from hospital and travel time) were associated with hidden costs. Hidden costs can be a

  2. Liposomal systems as viable drug delivery technology for skin cancer sites with an outlook on lipid-based delivery vehicles and diagnostic imaging inputs for skin conditions'.

    Science.gov (United States)

    Akhtar, Naseem; Khan, Riaz A

    2016-10-01

    Skin cancer is among one of the most common human malignancies wide-spread world-over with mortality statistics rising continuously at an alarming rate. The increasing frequency of these malignancies has marked the need for adopting effective treatment plan coupled with better and site-specific delivery options for the desired therapeutic agent's availability at the affected site. The concurrent delivery approaches to cancerous tissues are under constant challenge and, as a result, are evolving and gaining advancements in terms of delivery modes, therapeutic agents and site-specificity of the therapeutics delivery. The lipid-based liposomal drug delivery is an attractive and emerging option, and which is meticulously shaping up beyond a threshold level to a promising, and viable route for the effective delivery of therapeutic agents and other required injuctions to the skin cancer. An update on liposomal delivery of chemotherapeutic agents, natural-origin compounds, photosensitizer, and DNA repair enzymes as well as other desirable and typical delivery modes employed in drug delivery and in the treatment of skin cancers is discussed in details. Moreover, liposomal delivery of nucleic acid-based therapeutics, i.e., small interfering RNA (siRNA), mRNA therapy, and RGD-linked liposomes are among the other promising novel technology under constant development. The current clinical applicability, viable clinical plans, future prospects including transport feasibility of delivery vesicles and imaging techniques in conjunction with the therapeutic agents is also discussed. The ongoing innovations in liposomal drug delivery technology for skin cancers hold promise for further development of the methodology for better, more effective and site-specific delivery as part of the better treatment plan by ensuring faster drug transport, better and full payload delivery with enough and required concentration of the dose. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Polymer-peptide delivery platforms: effect of oligopeptide orientation on polymer-based DNA delivery.

    Science.gov (United States)

    Parelkar, Sangram S; Letteri, Rachel; Chan-Seng, Delphine; Zolochevska, Olga; Ellis, Jayne; Figueiredo, Marxa; Emrick, Todd

    2014-04-14

    The success of nonviral transfection using polymers hinges on efficient nuclear uptake of nucleic acid cargo and overcoming intra- and extracellular barriers. By incorporating PKKKRKV heptapeptide pendent groups as nuclear localization signals (NLS) on a polymer backbone, we demonstrate protein expression levels higher than those obtained from JetPEI and Lipofectamine 2000, the latter being notorious for coupling high transfection efficiency with cytotoxicity. The orientation of the NLS peptide grafts markedly affected transfection performance. Polymers with the sequence attached to the backbone from the valine residue achieved a level of nuclear translocation higher than the levels of those having the NLS groups attached in the opposite orientation. The differences in nuclear localization and DNA complexation strength between the two orientations correlated with a striking difference in protein expression, both in cell culture and in vivo. Polyplexes formed from these comb polymer structures exhibited transfection efficiencies superior to those of Lipofectamine 2000 but with greatly reduced toxicity. Moreover, these novel polymers, when administered by intramuscular ultrasound-mediated delivery, allowed a high level of reporter gene expression in mice, demonstrating their therapeutic promise in vivo.

  4. Erythrocytes-based synthetic delivery systems: transition from conventional to novel engineering strategies.

    Science.gov (United States)

    Bhateria, Manisha; Rachumallu, Ramakrishna; Singh, Rajbir; Bhatta, Rabi Sankar

    2014-08-01

    Erythrocytes (red blood cells [RBCs]) and artificial or synthetic delivery systems such as liposomes, nanoparticles (NPs) are the most investigated carrier systems. Herein, progress made from conventional approach of using RBC as delivery systems to novel approach of using synthetic delivery systems based on RBC properties will be reviewed. We aim to highlight both conventional and novel approaches of using RBCs as potential carrier system. Conventional approaches which include two main strategies are: i) directly loading therapeutic moieties in RBCs; and ii) coupling them with RBCs whereas novel approaches exploit structural, mechanical and biological properties of RBCs to design synthetic delivery systems through various engineering strategies. Initial attempts included coupling of antibodies to liposomes to specifically target RBCs. Knowledge obtained from several studies led to the development of RBC membrane derived liposomes (nanoerythrosomes), inspiring future application of RBC or its structural features in other attractive delivery systems (hydrogels, filomicelles, microcapsules, micro- and NPs) for even greater potential. In conclusion, this review dwells upon comparative analysis of various conventional and novel engineering strategies in developing RBC based drug delivery systems, diversifying their applications in arena of drug delivery. Regardless of the challenges in front of us, RBC based delivery systems offer an exciting approach of exploiting biological entities in a multitude of medical applications.

  5. Electroporation of DC-3F cells is a dual process.

    Science.gov (United States)

    Wegner, Lars H; Frey, Wolfgang; Silve, Aude

    2015-04-07

    Treatment of biological material by pulsed electric fields is a versatile technique in biotechnology and biomedicine used, for example, in delivering DNA into cells (transfection), ablation of tumors, and food processing. Field exposure is associated with a membrane permeability increase usually ascribed to electroporation, i.e., formation of aqueous membrane pores. Knowledge of the underlying processes at the membrane level is predominantly built on theoretical considerations and molecular dynamics (MD) simulations. However, experimental data needed to monitor these processes with sufficient temporal resolution are scarce. The whole-cell patch-clamp technique was employed to investigate the effect of millisecond pulsed electric fields on DC-3F cells. Cellular membrane permeabilization was monitored by a conductance increase. For the first time, to our knowledge, it could be established experimentally that electroporation consists of two clearly separate processes: a rapid membrane poration (transient electroporation) that occurs while the membrane is depolarized or hyperpolarized to voltages beyond so-called threshold potentials (here, +201 mV and -231 mV, respectively) and is reversible within ∼100 ms after the pulse, and a long-term, or persistent, permeabilization covering the whole voltage range. The latter prevailed after the pulse for at least 40 min, the postpulse time span tested experimentally. With mildly depolarizing or hyperpolarizing pulses just above threshold potentials, the two processes could be separated, since persistent (but not transient) permeabilization required repetitive pulse exposure. Conductance increased stepwise and gradually with depolarizing and hyperpolarizing pulses, respectively. Persistent permeabilization could also be elicited by single depolarizing/hyperpolarizing pulses of very high field strength. Experimental measurements of propidium iodide uptake provided evidence of a real membrane phenomenon, rather than a mere

  6. Engineering the Delivery System for CRISPR-Based Genome Editing.

    Science.gov (United States)

    Glass, Zachary; Lee, Matthew; Li, Yamin; Xu, Qiaobing

    2018-01-02

    Clustered regularly interspaced short palindromic repeat-CRISPR-associated protein (CRISPR-Cas) systems, found in nature as microbial adaptive immune systems, have been repurposed into an important tool in biological engineering and genome editing, providing a programmable platform for precision gene targeting. These tools have immense promise as therapeutics that could potentially correct disease-causing mutations. However, CRISPR-Cas gene editing components must be transported directly to the nucleus of targeted cells to exert a therapeutic effect. Thus, efficient methods of delivery will be critical to the success of therapeutic genome editing applications. Here, we review current strategies available for in vivo delivery of CRISPR-Cas gene editing components and outline challenges that need to be addressed before this powerful tool can be deployed in the clinic. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. The primipara respons based on individual personality type to the intensity of delivery pain

    Directory of Open Access Journals (Sweden)

    Gita N Sari

    2016-01-01

    Full Text Available Delivery period is one of periods that can cause stress to the mother and the fetus. This period is the natural common phenomenon that for some women subjectively can be considered as pain process that can cause simultaneous anxiety and pain. Psychology research has shown that pain is not only connected to physical respond, the culture that teaches and nurtures us also play important role in coping the pain. These two factors shape different personality for each individual. The objective of this study is to find out the primipara respons based on individual personality type to the intensity of delivery pain. The method of this study was analytical method with survey cross sectional approach. The data was collected prospectively from interview and questionnaire in the same time to find out the correlation between individual personality type and the intensity of delivery pain based on inclusive and exclusive period February 1st 2009 to April 30th 2009. The result with chi-square test and spearman rank test showed significant correlation between individual personality type and the intensity of delivery pain (X2= 8,571 ; p = 0,014. There is the negative correlation between extrovert individual personality and intensity of delivery pain (rs= -0,730; p <0,001, and there is the positive correlation between introvert individual personality type and intensity of delivery pain (rs = 0,726; p <0,001. Based on mann whitney, showed significant difference between extrovert personality type and introverts personality type to intensity of delivery pain (Z M-W: 3,050, p: 0,002. Based on chi-square test showed significant correlation between knowledge based on individual personality type to the intensity of delivery pain (X2= 4,418; p = 0,036 The conclusion of these study are the more extrovert individual personality type the less intense the delivery pain would be, the more introvert individual personality type then the more intense delivery pain would be. The

  8. Calcium electroporation induces tumor eradication, long-lasting immunity and cytokine responses in the CT26 colon cancer mouse model

    DEFF Research Database (Denmark)

    Falk, Hanne; Forde, Patrick F; Bay, Marie Lund

    2017-01-01

    Electroporation is used in cancer treatment because of its ability to increase local cytotoxicity of e.g. bleomycin (electrochemotherapy) and calcium (calcium electroporation). Calcium electroporation is a novel anticancer treatment that selectively kills cancer cells by necrosis, a cell death pa...

  9. Continuing progress toward controlled intracellular delivery of semiconductor quantum dots

    Science.gov (United States)

    Breger, Joyce; Delehanty, James B; Medintz, Igor L

    2015-01-01

    The biological applications of luminescent semiconductor quantum dots (QDs) continue to grow at a nearly unabated pace. This growth is driven, in part, by their unique photophysical and physicochemical properties which have allowed them to be used in many different roles in cellular biology including: as superior fluorophores for a wide variety of cellular labeling applications; as active platforms for assembly of nanoscale sensors; and, more recently, as a powerful tool to understand the mechanisms of nanoparticle mediated drug delivery. Given that controlled cellular delivery is at the intersection of all these applications, the latest progress in delivering QDs to cells is examined here. A brief discussion of relevant considerations including the importance of materials preparation and bioconjugation along with the continuing issue of endosomal sequestration is initially provided for context. Methods for the cellular delivery of QDs are then highlighted including those based on passive exposure, facilitated strategies that utilize peptides or polymers and fully active modalities such as electroporation and other mechanically based methods. Following on this, the exciting advent of QD cellular delivery using multiple or combined mechanisms is then previewed. Several recent methods reporting endosomal escape of QD materials in cells are also examined in detail with a focus on the mechanisms by which access to the cytosol is achieved. The ongoing debate over QD cytotoxicity is also discussed along with a perspective on how this field will continue to evolve in the future. PMID:25154379

  10. Transfection of small numbers of human endothelial cells by electroporation and synthetic amphiphiles

    NARCIS (Netherlands)

    van Leeuwen, E B; van der Veen, A Y; Hoekstra, D; Engberts, J B; Halie, M R; van der Meer, J; Ruiters, M H

    OBJECTIVES: This study compared the efficiency of electroporation and synthetic amphiphiles. (SAINT-2pp/DOPE) in transfecting small numbers of human endothelial cells. METHODS AND RESULTS: Optimal transfection conditions were tested and appeared to be 400 V and 960 microF for electroporation and a

  11. A primary current distribution model of a novel micro-electroporation channel configuration.

    Science.gov (United States)

    Troszak, Gregory D; Rubinsky, Boris

    2010-10-01

    Traditional macro and micro-electroporation devices utilize facing electrodes, which generate electric fields inversely proportional to their separation distance. Although the separation distances in micro-electroporation devices are significantly smaller than those in macro-electroporation devices, they are limited by cell size. Because of this, significant potential differences are required to induce electroporation. These potential differences are often large enough to cause water electrolysis, resulting in electrode depletion and bubble formation, both of which adversely affect the electroporation process. Here, we present a theoretical study of a novel micro-electroporation channel composed of an electrolyte flowing over a series of adjacent electrodes separated by infinitesimally small insulators. Application of a small, non-electrolysis inducing potential difference between the adjacent electrodes results in radially-varying electric fields that emanate from these insulators, causing cells flowing through the channel to experience a pulsed electric field. This eliminates the need for a pulse generator, making a minimal power source (such as a battery) the only electrical equipment that is needed. A non-dimensional primary current distribution model of the novel micro-electroporation channel shows that decreasing the channel height results in an exponential increase in the electric field magnitude, and that cells experience exponentially greater electric field magnitudes the closer they are to the channel walls. Finally, dimensional primary current distribution models of two potential applications, water sterilization and cell transfection, demonstrate the practical feasibility of the novel micro-electroporation channel.

  12. Direct therapeutic applications of calcium electroporation to effectively induce tumor necrosis

    DEFF Research Database (Denmark)

    Frandsen, Stine Krog; Gissel, Hanne; Hojman, Pernille

    2012-01-01

    in vivo. Calcium electroporation elicited dramatic antitumor responses in which 89% of treated tumors were eliminated. Histologic analyses indicated complete tumor necrosis. Mechanistically, calcium electroporation caused acute ATP depletion likely due to a combination of increased cellular use of ATP...... access to this technology for many cancer patients in developed and developing countries....

  13. Targeted electroporation of defined lateral ventricular walls: a novel and rapid method to study fate specification during postnatal forebrain neurogenesis

    Science.gov (United States)

    2011-01-01

    Background Postnatal olfactory bulb (OB) neurogenesis involves the generation of granule and periglomerular cells by neural stem cells (NSCs) located in the walls of the lateral ventricle (LV). Recent studies show that NSCs located in different regions of the LV give rise to different types of OB neurons. However, the molecular mechanisms governing neuronal specification remain largely unknown and new methods to approach these questions are needed. Results In this study, we refine electroporation of the postnatal forebrain as a technique to perform precise and accurate delivery of transgenes to NSCs located in distinct walls of the LV in the mouse. Using this method, we confirm and expand previous studies showing that NSCs in distinct walls of the LV produce neurons that invade different layers of the OB. Fate mapping of the progeny of radial glial cells located in these distinct LV walls reveals their specification into defined subtypes of granule and periglomerular neurons. Conclusions Our results provide a baseline with which future studies aiming at investigating the role of factors in postnatal forebrain neuronal specification can be compared. Targeted electroporation of defined LV NSC populations will prove valuable to study the genetic factors involved in forebrain neuronal specification. PMID:21466691

  14. Assembling nanoparticle coatings to improve the drug delivery performance of lipid based colloids

    Science.gov (United States)

    Simovic, Spomenka; Barnes, Timothy J.; Tan, Angel; Prestidge, Clive A.

    2012-02-01

    Lipid based colloids (e.g. emulsions and liposomes) are widely used as drug delivery systems, but often suffer from physical instabilities and non-ideal drug encapsulation and delivery performance. We review the application of engineered nanoparticle layers at the interface of lipid colloids to improve their performance as drug delivery systems. In addition we focus on the creation of novel hybrid nanomaterials from nanoparticle-lipid colloid assemblies and their drug delivery applications. Specifically, nanoparticle layers can be engineered to enhance the physical stability of submicron lipid emulsions and liposomes, satbilise encapsulated active ingredients against chemical degradation, control molecular transport and improve the dermal and oral delivery characteristics, i.e. increase absorption, bioavailability and facilitate targeted delivery. It is feasible that hybrid nanomaterials composed of nanoparticles and colloidal lipids are effective encapsulation and delivery systems for both poorly soluble drugs and biological drugs and may form the basis for the next generation of medicines. Additional pre-clinical research including specific animal model studies are required to advance the peptide/protein delivery systems, whereas the silica lipid hybrid systems have now entered human clinical trials for poorly soluble drugs.

  15. PEG-based degradable networks for drug delivery applications

    Science.gov (United States)

    Ostroha, Jamie L.

    The controlled delivery of therapeutic agents by biodegradable hydrogels has become a popular mechanism for drug administration in recent years. Hydrogels are three-dimensional networks of polymer chains held together by crosslinks. Although the changes which the hydrogel undergoes in solution are important to a wide range of experimental studies, they have not been investigated systematically and the factors which influence the degree of swelling have not been adequately described. Hydrogels made of poly(ethylene glycol) (PEG) will generally resist degradation in aqueous conditions, while a hydrogel made from a copolymer of poly(lactic acid) (PLA) and PEG will degrade via hydrolysis of the lactic acid group. This ability to degrade makes these hydrogels promising candidates for controlled release drug delivery systems. The goal of this research was to characterize the swelling and degradation of both degradable and non-degradable gels and to evaluate the release of different drugs from these hydrogels, where the key variable is the molecular weight of the PEG segment. These hydrogels were formed by the addition and subsequent chemically crosslinking of methacrylate end groups. During crosslinking, both PEG and LA-PEG-LA hydrogels of varied PEG molecular weight were loaded with Vitamin B12, Insulin, Haloperidol, and Dextran. It was shown that increasing PEG molecular weight produces a hydrogel with larger pores, thus increasing water uptake and degradation rate. While many environmental factors do not affect the swelling behavior, they do significantly impact the degradation of the hydrogel, and thus the release of incorporated therapeutic agents.

  16. Skin Delivery of Kojic Acid-Loaded Nanotechnology-Based Drug Delivery Systems for the Treatment of Skin Aging

    Directory of Open Access Journals (Sweden)

    M. L. Gonçalez

    2013-01-01

    Full Text Available The aging process causes a number of changes in the skin, including oxidative stress and dyschromia. The kojic acid (KA is iron chelator employed in treatment of skin aging, and inhibits tyrosinase, promotes depigmentation. Nanotechnology-based drug delivery systems, such as liquid crystalline systems (LCSs, can modulate drug permeation through the skin and improve the drug activity. This study is aimed at structurally developing and characterizing a kojic acid-loaded LCS, consists of water (W, cetostearyl isononanoate (oil—O and PPG-5-CETETH-20 (surfactant-S and evaluating its in vitro skin permeation and retention. Three regions of the diagram were selected for characterization: A (35% O, 50% S, 15% W, B (30% O, 50% S, 20% W and C (20% O, 50% S, 30% W, to which 2% KA was added. The formulations were subjected to polarized light microscopy, which indicated the presence of a hexagonal mesophase. Texture and bioadhesion assay showed that formulation B is suitable for topical application. According to the results from the in vitro permeation and retention of KA, the formulations developed can modulate the permeation of KA in the skin. The in vitro cytotoxic assays showed that KA-unloaded LCS and KA-loaded LCS didn't present cytotoxicity. PPG-5-CETETH-20-based systems may be a promising platform for KA skin delivery.

  17. Nonlinear electro-mechanobiological behavior of cell membrane during electroporation

    KAUST Repository

    Deng, Peigang

    2012-01-01

    A nonlinear electroporation (EP) model is proposed to study the electro-mechanobiological behavior of cell membrane during EP, by taking the nonlinear large deformation of the membrane into account. The proposed model predicts the critical transmembrane potential and the activation energy for EP, the equilibrium pore size, and the resealing process of the pore. Single-cell EP experiments using a micro EP chip were conducted on chicken red blood cells at different temperatures to determine the activation energy and the critical transmembrane potential for EP. The experimental results are in good agreement with the theoretical predictions. © 2012 American Institute of Physics.

  18. Improved brain uptake of peptide-based CNS drugs via alternative routes of administrations of its nanocarrier delivery systems: a promising strategy for CNS targeting delivery of peptides.

    Science.gov (United States)

    Qian, Shuai; Wang, Qianwen; Zuo, Zhong

    2014-11-01

    Recently, developing peptide-based drugs to treat CNS diseases has gained increasing attention in both academics and pharmaceutical industry. However, targeting delivery of peptides to brain is one of the most challenging problems faced in the treatment of CNS diseases. After explaining the brain barriers limiting the delivery of peptides, the current review focuses on summarizing the most promising approaches for the enhancement of peptide and brain uptake, including delivery via alternative routes of administrations or using nanocarriers or intranasal administration of nanocarriers loaded with peptide. In addition, the biopharmaceutic, pharmacokinetic and pharmacodynamic details of several successful peptide-based CNS drugs are highlighted. Although using nanocarriers or delivery via alternative routes could improve to a certain extent the brain uptake of peptides, the magnitude of changes remains moderate. Alternatively, intranasal administration of nanocarriers loaded with peptide has been demonstrated to be an effective approach for CNS-targeted delivery of peptides.

  19. Enzymatically triggered multifunctional delivery system based on hyaluronic acid micelles

    KAUST Repository

    Deng, Lin

    2012-01-01

    Tumor targetability and stimuli responsivity of drug delivery systems (DDS) are key factors in cancer therapy. Implementation of multifunctional DDS can afford targetability and responsivity at the same time. Herein, cholesterol molecules (Ch) were coupled to hyaluronic acid (HA) backbones to afford amphiphilic conjugates that can self-assemble into stable micelles. Doxorubicin (DOX), an anticancer drug, and superparamagnetic iron oxide (SPIO) nanoparticles (NPs), magnetic resonance imaging (MRI) contrast agents, were encapsulated by Ch-HA micelles and were selectively released in the presence of hyaluronidase (Hyals) enzyme. Cytotoxicity and cell uptake studies were done using three cancer cell lines (HeLa, HepG2 and MCF7) and one normal cell line (WI38). Higher Ch-HA micelles uptake was seen in cancer cells versus normal cells. Consequently, DOX release was elevated in cancer cells causing higher cytotoxicity and enhanced cell death. © 2012 The Royal Society of Chemistry.

  20. Nanotechnology-based combinational drug delivery: an emerging approach for cancer therapy.

    Science.gov (United States)

    Parhi, Priyambada; Mohanty, Chandana; Sahoo, Sanjeeb Kumar

    2012-09-01

    Combination therapy for the treatment of cancer is becoming more popular because it generates synergistic anticancer effects, reduces individual drug-related toxicity and suppresses multi-drug resistance through different mechanisms of action. In recent years, nanotechnology-based combination drug delivery to tumor tissues has emerged as an effective strategy by overcoming many biological, biophysical and biomedical barriers that the body stages against successful delivery of anticancer drugs. The sustained, controlled and targeted delivery of chemotherapeutic drugs in a combination approach enhanced therapeutic anticancer effects with reduced drug-associated side effects. In this article, we have reviewed the scope of various nanotechnology-based combination drug delivery approaches and also summarized the current perspective and challenges facing the successful treatment of cancer. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Designing a Mobile Based Library Information and Service Delivery System: A Model Plan for Libraries of Bangladesh

    National Research Council Canada - National Science Library

    Elahi, Md. Hasinul

    2016-01-01

    ... with the help of mobile phones and their applications, design a mobile based library information and service delivery system, and identify major challenges regarding mobile based library information...

  2. High-molecular-weight HPMA-based polymer drug carriers for delivery to tumor.

    Science.gov (United States)

    Kostka, L; Etrych, T

    2016-10-20

    In this work, design and synthesis of high-molecular-weight N-(2-hydroxypropyl)methacrylamide-based polymer drug delivery systems tailored for cancer therapy is summarized. Moreover, the influence of their architecture on tumor accumulation and in vivo anti-cancer efficacy is discussed. Mainly, the high-molecular-weight delivery systems, such as branched, grafted, multi-block, star-like or micellar systems, with molecular weights greater than the renal threshold are discussed and reviewed in detail.

  3. New method for electroporation of Lactobacillus species grown in high salt.

    Science.gov (United States)

    Palomino, Maria Mercedes; Allievi, Mariana C; Prado-Acosta, Mariano; Sanchez-Rivas, Carmen; Ruzal, Sandra M

    2010-11-01

    We here describe a new method for electroporation of Lactobacillus species, obligately homofermentative and facultatively heterofermentative, based on the cell-wall weakening resulting from growth in high-salt media. For L. casei, optimum transformation efficiency of up to 10(5) transformants per microgram of plasmid DNA was achieved following growth in the presence of 0.9 M NaCl. Plasmids of different sizes and replication origins were also similarly transformed. These competent cells could be used either directly or stored frozen, up to 1 month, for future use, with similar efficiency. This protocol was assayed with different Lactobacillus species: L. delbrueckii subsp. lactis, L. paracasei, L. plantarum and L. acidophilus, and it was found that they were transformed with similar efficiency. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. The Effect of Blood Flow on Magnetic Resonance Imaging of Non Thermal Irreversible Electroporation

    Science.gov (United States)

    Hjouj, Mohammad; Lavee, Jacob; Last, David; Guez, David; Daniels, Dianne; Sharabi, Shirley; Rubinsky, Boris; Mardor, Yael

    2013-10-01

    To generate an understanding of the physiological significance of MR images of Non-Thermal Irreversible Electroporation (NTIRE) we compared the following MR imaging sequences: T1W, T2W, PD, GE, and T2 SPAIR acquired after NTIRE treatment in a rodent liver model. The parameters that were studied included the presence or absence of a Gd-based contrast agent, and in vivo and ex-vivo NTIRE treatments in the same liver. NTIRE is a new minimally invasive tissue ablation modality in which pulsed electric fields cause molecularly selective cell death while, the extracellular matrix and large blood vessels remain patent. This attribute of NTIRE is of major clinical importance as it allows treatment of undesirable tissues near critical blood vessels. The presented study results suggest that MR images acquired following NTIRE treatment are all directly related to the unique pattern of blood flow after NTIRE treatment and are not produced in the absence of blood flow.

  5. Local Ablative Strategies for Ductal Pancreatic Cancer (Radiofrequency Ablation, Irreversible Electroporation): A Review

    Science.gov (United States)

    Paiella, Salvatore; Salvia, Roberto; Ramera, Marco; Girelli, Roberto; Frigerio, Isabella; Giardino, Alessandro; Allegrini, Valentina; Bassi, Claudio

    2016-01-01

    Pancreatic ductal adenocarcinoma (PDAC) has still a dismal prognosis. Locally advanced pancreatic cancer (LAPC) accounts for the 40% of the new diagnoses. Current treatment options are based on chemo- and radiotherapy regimens. Local ablative techniques seem to be the future therapeutic option for stage-III patients with PDAC. Radiofrequency Ablation (RFA) and Irreversible Electroporation (IRE) are actually the most emerging local ablative techniques used on LAPC. Initial clinical studies on the use of these techniques have already demonstrated encouraging results in terms of safety and feasibility. Unfortunately, few studies on their efficacy are currently available. Even though some reports on the overall survival are encouraging, randomized studies are still required to corroborate these findings. This study provides an up-to-date overview and a thematic summary of the current available evidence on the application of RFA and IRE on PDAC, together with a comparison of the two procedures. PMID:26981115

  6. Biodegradable polycarbonate-based stimuli-responsive nanosystems for intracellular drug delivery

    NARCIS (Netherlands)

    Chen, Wei

    2013-01-01

    Biodegradable nanosystems based on functional polycarbonate-based polymers have propelled the development of targeted and controlled drug and gene delivery. Functional polycarbonate-based polymers have overcome many drawbacks which limited the application of the common polyester materials in this

  7. Magnetic Nanomaterials for Hyperthermia-based Therapy and Controlled Drug Delivery

    Science.gov (United States)

    Kumar, Challa S. S. R.; Mohammad, Faruq

    2011-01-01

    Previous attempts to review the literature on magnetic nanomaterials for hyperthermia-based therapy focused primarily on magnetic fluid hyperthermia (MFH) using mono metallic/metal oxide nanoparticles. The term “Hyperthermia” in the literature was also confined only to include use of heat for therapeutic applications. Recently, there have been a number of publications demonstrating magnetic nanoparticle-based hyperthermia to generate local heat resulting in the release of drugs either bound to the magnetic nanoparticle or encapsulated within polymeric matrices. In this review article, we present a case for broadening the meaning of the term “hyperthermia” by including thermotherapy as well as magnetically modulated controlled drug delivery. We provide a classification for controlled drug delivery using hyperthermia: Hyperthermia-based controlled Drug delivery through Bond Breaking (DBB) and Hyperthermia-based controlled Drug delivery through Enhanced Permeability (DEP). The review also covers, for the first time, core-shell type magnetic nanomaterials, especially nanoshells prepared using layer-by-layer self-assembly, for the application of hyperthermia-based therapy and controlled drug delivery. The highlight of the review article is to portray potential opportunities in the combination of hyperthermia-based therapy and controlled drug release paradigms for successful application in personalized medicine. PMID:21447363

  8. Magnetic nanomaterials for hyperthermia-based therapy and controlled drug delivery.

    Science.gov (United States)

    Kumar, Challa S S R; Mohammad, Faruq

    2011-08-14

    Previous attempts to review the literature on magnetic nanomaterials for hyperthermia-based therapy focused primarily on magnetic fluid hyperthermia (MFH) using mono metallic/metal oxide nanoparticles. The term "hyperthermia" in the literature was also confined only to include use of heat for therapeutic applications. Recently, there have been a number of publications demonstrating magnetic nanoparticle-based hyperthermia to generate local heat resulting in the release of drugs either bound to the magnetic nanoparticle or encapsulated within polymeric matrices. In this review article, we present a case for broadening the meaning of the term "hyperthermia" by including thermotherapy as well as magnetically modulated controlled drug delivery. We provide a classification for controlled drug delivery using hyperthermia: Hyperthermia-based controlled drug delivery through bond breaking (DBB) and hyperthermia-based controlled drug delivery through enhanced permeability (DEP). The review also covers, for the first time, core-shell type magnetic nanomaterials, especially nanoshells prepared using layer-by-layer self-assembly, for the application of hyperthermia-based therapy and controlled drug delivery. The highlight of the review article is to portray potential opportunities for the combination of hyperthermia-based therapy and controlled drug release paradigms--towards successful application in personalized medicine. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Nanoparticle-Based Delivery System for Biomedical Applications of RNAi

    DEFF Research Database (Denmark)

    Yang, Chuanxu

    RNA interference (RNAi) is a post-transcriptional gene silencing process triggered by double-strand RNA, including synthetic short interfering RNA (siRNA) and endogenous microRNA (miRNA). RNAi has attracted great attention for developing a new class of therapeutics, due to its capability to speci......RNA and transport them to the action site in the target cells. This thesis describes the development of various nanocarriers for siRNA/miRNA delivery and investigate their potential biomedical applications including: anti-inflammation, tissue engineering and cancer......RNA interference (RNAi) is a post-transcriptional gene silencing process triggered by double-strand RNA, including synthetic short interfering RNA (siRNA) and endogenous microRNA (miRNA). RNAi has attracted great attention for developing a new class of therapeutics, due to its capability...... to specifically inhibit the expression of virtually any gene with high efficiency, including the “undruggable” targets. To activate the RNAi pathway, siRNA or miRNA molecules must be transported into the cytoplasm of target cells. However, various barriers impede the transportation of these fragile macromolecules...

  10. The role of pH fronts in reversible electroporation.

    Directory of Open Access Journals (Sweden)

    Pablo Turjanski

    Full Text Available We present experimental measurements and theoretical predictions of ion transport in agar gels during reversible electroporation (ECT for conditions typical to many clinical studies found in the literature, revealing the presence of pH fronts emerging from both electrodes. These results suggest that pH fronts are immediate and substantial. Since they might give rise to tissue necrosis, an unwanted condition in clinical applications of ECT as well as in irreversible electroporation (IRE and in electrogenetherapy (EGT, it is important to quantify their extent and evolution. Here, a tracking technique is used to follow the space-time evolution of these pH fronts. It is found that they scale in time as t(½, characteristic of a predominantly diffusive process. Comparing ECT pH fronts with those arising in electrotherapy (EChT, another treatment applying constant electric fields whose main goal is tissue necrosis, a striking result is observed: anodic acidification is larger in ECT than in EChT, suggesting that tissue necrosis could also be greater. Ways to minimize these adverse effects in ECT are suggested.

  11. Conditions for transformation of Pasteurella multocida by electroporation.

    Science.gov (United States)

    Jablonski, L; Sriranganathan, N; Boyle, S M; Carter, G R

    1992-01-01

    Conditions for electroporation of plasmid DNA into Pasteurella multocida were determined for use in developing a cloning system to study virulence factors of P. multocida. The highest efficiency of transformation (1.25 x 10(7) cfu/micrograms DNA) was obtained when 7.6 x 10(10) cells of P. multocida strain R473 were electroporated at 12.5 kV/cm (10 ms, 5 ng of pVM109). Transformation efficiencies of cells prepared at mid-log-phase were approximately 0.5 log10 lower than early, late, or stationary phases. Neither pBR322 nor pUC-19 were able to transform strain R473 under these conditions, even when DNA concentrations were increased to 1 microgram. When pBR322 was ligated with a Pasteurella plasmid, pLAR-1, the hybrid was able to transform strain R473 at an efficiency between 4.5 x 10(2) and 8 x 10(4) cfu/micrograms DNA. Six strains of P. multocida including serotypes A, B, D, and E were transformed successfully.

  12. Percutaneous Irreversible Electroporation of a Large Centrally Located Hepatocellular Adenoma in a Woman with a Pregnancy Wish

    Energy Technology Data Exchange (ETDEWEB)

    Scheffer, Hester J., E-mail: hj.scheffer@vumc.nl; Melenhorst, Marleen C. A. M., E-mail: m.melenhorst@vumc.nl; Tilborg, Aukje A. J. M. van, E-mail: a.vantilborg@vumc.nl [VU University Medical Center, Department of Radiology and Nuclear Medicine (Netherlands); Nielsen, Karin, E-mail: k.nielsen@vumc.nl [Department of Surgery (Netherlands); Nieuwkerk, Karin M. van, E-mail: cmj.vannieuwkerk@vumc.nl; Vries, Richard A. de, E-mail: ra.devries@vumc.nl [VU University Medical Center, Department of Gastroenterology and Hepatology (Netherlands); Tol, Petrousjka van den [Department of Surgery (Netherlands); Meijerink, Martijn R., E-mail: mr.meijerink@vumc.nl [VU University Medical Center, Department of Radiology and Nuclear Medicine (Netherlands)

    2015-08-15

    Irreversible electroporation (IRE) is a novel image-guided ablation technique that is rapidly gaining popularity in the treatment of malignant liver tumors located near large vessels or bile ducts. We describe a 28-year-old female patient with a 5 cm large, centrally located hepatocellular adenoma who wished to get pregnant. Regarding the risk of growth and rupture of the adenoma caused by hormonal changes during pregnancy, treatment of the tumor was advised prior to pregnancy. However, due to its central location, the tumor was considered unsuitable for resection and thermal ablation. Percutaneous CT-guided IRE was performed without complications and led to rapid and impressive tumor shrinkage. Subsequent pregnancy and delivery went uncomplicated. This case report suggests that the indication for IRE may extend to the treatment of benign liver tumors that cannot be treated safely otherwise.

  13. Chitosan-based delivery systems for protein therapeutics and antigens

    NARCIS (Netherlands)

    Amidi, M.; Mastrobattista, E.; Jiskoot, W.; Hennink, W.E.

    Therapeutic peptides/proteins and protein-based antigens are chemically and structurally labile compounds, which are almost exclusively administered by parenteral injections. Recently, non-invasive mucosal routes have attracted interest for administration of these biotherapeutics. Chitosan-based

  14. Optimization of a plasmid electroporation protocol for Aeromonas salmonicida subsp. salmonicida.

    Science.gov (United States)

    Dallaire-Dufresne, Stéphanie; Emond-Rheault, Jean-Guillaume; Attéré, Sabrina A; Tanaka, Katherine H; Trudel, Mélanie V; Frenette, Michel; Charette, Steve J

    2014-03-01

    Aeromonas salmonicida subsp. salmonicida is a major fish pathogen. Molecular tools are required to study the virulence and genomic stability of this bacterium. An efficient electroporation-mediated transformation protocol for A. salmonicida subsp. salmonicida would make genetic studies faster and easier. In the present study, we designed the 4.1-kb pSDD1 plasmid as a tool for optimizing an electroporation protocol for A. salmonicida subsp. salmonicida. We systematically tested the electroporation conditions to develop a protocol that generates the maximum number of transformants. Under these optimal conditions (25 kV/cm, 200 Ω, 25 μF), we achieved an electroporation efficiency of up to 1×10(5) CFU/μg DNA. The electroporation protocol was also tested using another plasmid of 10.6-kb and three different strains of A. salmonicida subsp. salmonicida. The strains displayed significant differences in their electro-transformation competencies. Strain 01-B526 was the easiest to electroporate, especially with the pSDD1 plasmid. This plasmid was stably maintained in the 01-B526 transformants, as were the native plasmids, but could be easily cured by removing the selection conditions. This is the first efficient electroporation protocol reported for A. salmonicida subsp. salmonicida, and offers new possibilities for studying this bacterium. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Breech at term--mode of delivery? A register-based study

    DEFF Research Database (Denmark)

    Krebs, L; Langhoff-Roos, J; Weber, Tom

    1995-01-01

    . Register-based cohort study of all (n = 15718) singleton term breech deliveries of non-malformed infants in Denmark 1982-1990. Process and outcome measures: mode of delivery, gestational age, birth weight, congenital malformations, intrapartum death, Apgar scores and early neonatal death. RESULTS. A total...... of 3247 (20.7%) term infants were delivered vaginally, 7106 (45.3%) by elective and 5356 (34.1%) by emergency cesarean section. Infants delivered vaginally and by emergency cesarean section had significantly higher rates of mortality (intrapartum and early neonatal death) and morbidity (low Apgar scores......) when compared to those delivered by elective cesarean section. In vaginal deliveries, parity was not correlated with outcome, but infants with a birth weight above 4000 grams had significantly higher rates of low Apgar scores. CONCLUSIONS. Register data on singleton term breech deliveries imply...

  16. Recent Advances in the Synthesis of Graphene-Based Nanomaterials for Controlled Drug Delivery

    Directory of Open Access Journals (Sweden)

    Zhuqing Wang

    2017-11-01

    Full Text Available Graphene-based nanomaterials have exhibited wide applications in nanotechnology, materials science, analytical science, and biomedical engineering due to their unique physical and chemical properties. In particular, graphene has been an excellent nanocarrier for drug delivery application because of its two-dimensional structure, large surface area, high stability, good biocompatibility, and easy surface modification. In this review, we present the recent advances in the synthesis and drug delivery application of graphene-based nanomaterials. The modification of graphene and the conjugation of graphene with other materials, such as small molecules, nanoparticles, polymers, and biomacromolecules as functional nanohybrids are introduced. In addition, the controlled drug delivery with the fabricated graphene-based nanomaterials are demonstrated in detail. It is expected that this review will guide the chemical modification of graphene for designing novel functional nanohybrids. It will also promote the potential applications of graphene-based nanomaterials in other biomedical fields, like biosensing and tissue engineering.

  17. Cervical dilation at time of caesarean delivery in nulliparous women: a population-based cohort study.

    Science.gov (United States)

    Riddell, C A; Kaufman, J S; Strumpf, E C; Abenhaim, H A; Hutcheon, J A

    2017-10-01

    Our objective was to describe contemporary practice patterns in the timing of caesarean delivery in relation to cervical dilation, overall and by indication for caesarean. Our secondary objective was to examine how commonly caesarean delivery was performed for labour dystocia at dilations below 4 cm or without the use of oxytocin, overall and between hospitals. Retrospective, population-based cohort study. Ontario, Alberta, and British Columbia, Canada, 2008-2012. Nulliparous women in labour who delivered term singletons in cephalic position. Histograms were used to examine the distribution of cervical dilation at time of caesarean delivery, overall and by indication for caesarean. Funnel plots were used to illustrate variation in hospital-level rates of caesarean deliveries for labour dystocia that were performed early (dilation) or without the use of oxytocin. Cervical dilation (in centimetres) at time of caesarean delivery. The population-based cohort comprised 392 025 women, of whom 18.8% had a caesarean delivery. Of first-stage caesareans for labour dystocia in women who entered labour spontaneously, 13.6% (95% CI 12.9, 14.2) had dilations dilation or without oxytocin varies substantially across hospitals and suggests the need for institutions to review their practices and ensure that management of labour practice guidelines are followed. Many caesareans for labour dystocia are performed early during labour (dilation) or without oxytocin. © 2016 Royal College of Obstetricians and Gynaecologists.

  18. Direct Gene Transfer into Plant Mature Seeds via Electroporation After Vacuum Treatment

    Science.gov (United States)

    Hagio, Takashi

    A number of direct gene transfer methods have been used successfully in plant genetic engineering, providing powerful tools to investigate fundamental and applied problems in plant biology (Chowrira et al., 1996; D'halluin et al., 1992; Morandini and Salamini, 2003; Rakoczy-Trojanowska, 2002; Songstad et al., 1995). In cereals, several methods have been found to be suitable for obtaining transgenic plant; these include bombardment of scutellum (Hagio et al., 1995) and inflorescence cultures (He et al., 2001), and silicon carbide fiber-mediated DNA delivery (Asano et al., 1991) and Agrobacterium tumefaciens transformation (Potrykus, 1990). Electroporation of cereal protoplasts also has proved successful but it involves prolonged cell treatments and generally is limited by the difficulties of regeneration from cereal protoplast cultures (Fromm et al., 1987). Many laboratories worldwide are now using Agrobacterium as a vehicle for routine production of transgenic crop plants. The primary application of the particle system (Klein et al., 1987) has been for transformation of species recalcitrant to conventional Agrobacterium (Binns, 1990) or protoplast methods. But these conventional methods can be applied to the species and varieties that are amenable to tissue culture (Machii et al., 1998). Mature seeds are readily available and free from the seasonal limits that immature embryo, inflorescence, and anther have. This method enables us to produce transgenic plants without time-consuming tissue culture process.

  19. Effect of Thermal Gradients Created by Electromagnetic Fields on Cell-Membrane Electroporation Probed by Molecular-Dynamics Simulations

    Science.gov (United States)

    Song, J.; Garner, A. L.; Joshi, R. P.

    2017-02-01

    The use of nanosecond-duration-pulsed voltages with high-intensity electric fields (˜100 kV /cm ) is a promising development with many biomedical applications. Electroporation occurs in this regime, and has been attributed to the high fields. However, here we focus on temperature gradients. Our numerical simulations based on molecular dynamics predict the formation of nanopores and water nanowires, but only in the presence of a temperature gradient. Our results suggest a far greater role of temperature gradients in enhancing biophysical responses, including possible neural stimulation by infrared lasers.

  20. Erythrocyte-based drug delivery in Transfusion Medicine: Wandering questions seeking answers.

    Science.gov (United States)

    Tzounakas, Vassilis L; Karadimas, Dimitrios G; Papassideri, Issidora S; Seghatchian, Jerard; Antonelou, Marianna H

    2017-08-01

    Red blood cells (RBCs) represent the most commonly used and best-studied natural carriers in the history of drug delivery. Their abundance and long circulation half-life, their great immune-biocompatibility and biodegradability profiles, along with the availability of well established protocols for their safe collection, ex vivo processing and quality control make them advantageous as drug delivery systems (DDS). As a result, several drug-loading techniques (including encapsulation and surface conjugation) have been developed in order to construct RBC-based or RBC-inspired drug delivery vehicles for the effective treatment of infections, cancer, chronic and autoimmune diseases in both pre-clinical protocols and clinical trials. Despite the fact that the collected laboratory (in vitro and in vivo) and clinical data exhibit variable potential for translation into transfusion-associated prototypes and feasible protocols with significant clinical impact, little is known and done in the direction of drug delivery through RBC transfusion. Accordingly, several wandering questions for the application and utility of RBC-based drug delivery in transfusion medicine seek answers. By focusing on the most prominent of them, namely, "why not the stored/transfused RBCs", this review quotes some thoughtful considerations based on the current applications of RBCs as DDS, and on the potential application of RBC-based DDS in transfusion therapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. ELECTROPORATION OF CONFLUENT HM-1 ES CELLS LEADS TO HIGHER AMOUNT COLONIES

    Directory of Open Access Journals (Sweden)

    A.ALENA BENCSIK

    2008-05-01

    Full Text Available Electroporation is used to induce homologous recombination in the genome of the murine ES (embryonic stem cells. Routinelly subconfluent ES cells are recommended to be used in such experiments. Electroporation of immunoglobulin specific targeting vectors with different length of homology leads to reduced number of selected colonies. The enrichment of double selected colonies is high and thus the amount of HM-1 ES cell colonies for the analysis is very low. Here we show that the electroporation of confluent HM-1 ES cells leads to an increased amount of simple and double selected colonies.

  2. Electrospun fish protein fibers as a biopolymer-based carrier – implications for oral protein delivery

    DEFF Research Database (Denmark)

    Boutrup Stephansen, Karen; García-Díaz, María; Jessen, Flemming

    2014-01-01

    Purpose: Protein-based electrospun fibers have emerged as novel nanostructured materials for tissue engineering and drug delivery due to their unique structural characteristics, biocompatibility and biodegradability. The aim of this study was to explore the use of electrospun fibers based on fish...

  3. Charge-reversal Lipids, Peptide-based Lipids, and Nucleoside-based Lipids for Gene Delivery

    Science.gov (United States)

    LaManna, Caroline M.; Lusic, Hrvoje; Camplo, Michel; McIntosh, Thomas J.; Barthélémy, Philippe; Grinstaff, Mark W.

    2013-01-01

    Conspectus Twenty years after gene therapy was introduced in the clinic, advances in the technique continue to garner headlines as successes pique the interest of clinicians, researchers, and the public. Gene therapy’s appeal stems from its potential to revolutionize modern medical therapeutics by offering solutions to a myriad of diseases by tailoring the treatment to a specific individual’s genetic code. Both viral and non-viral vectors have been used in the clinic, but the low transfection efficiencies when utilizing non-viral vectors have lead to an increased focus on engineering new gene delivery vectors. To address the challenges facing non-viral or synthetic vectors, specifically lipid-based carriers, we have focused on three main themes throughout our research: 1) that releasing the nucleic acid from the carrier will increase gene transfection; 2) that utilizing biologically inspired designs, such as DNA binding proteins, to create lipids with peptide-based headgroups will improve delivery; and 3) that mimicking the natural binding patterns observed within DNA, by using lipids having a nucleoside headgroup, will give unique supramolecular assembles with high transfection efficiency. The results presented in this Account demonstrate that cellular uptake and transfection efficacy can be improved by engineering the chemical components of the lipid vectors to enhance nucleic acid binding and release kinetics. Specifically, our research has shown that the incorporation of a charge-reversal moiety to initiate change of the lipid from positive to negative net charge during the transfection process improves transfection. In addition, by varying the composition of the spacer (rigid, flexible, short, long, and aromatic) between the cationic headgroup and the hydrophobic chains, lipids can be tailored to interact with different nucleic acids (DNA, RNA, siRNA) and accordingly affect delivery, uptake outcomes, and transfection efficiency. Introduction of a peptide

  4. Recent development of chitosan-based polyelectrolyte complexes with natural polysaccharides for drug delivery.

    Science.gov (United States)

    Luo, Yangchao; Wang, Qin

    2014-03-01

    Chitosan, as a unique positively charged polysaccharide, has been one of the most popular biopolymers for development of drug delivery systems for various applications, due to its promising properties, including high biocompatibility, excellent biodegradability, low toxicity, as well as abundant availability and low production cost. Since last decade, increasing attention has been attracted by delivery systems fabricated from natural biopolymer-based polyelectrolyte complexes (PEC), formed by electrostatic interactions between two oppositely charged biopolymers. In order to tailor specific applications of chitosan-based PEC drug delivery systems, various forms have been developed in recent years, including nanoparticles, microparticles, beads, tablets, gels, as well as films and membranes. The present review focuses on the recent advances in drug delivery applications of chitosan-based PEC with other natural polysaccharides, including alginate, hyaluronic acid, pectin, carrageenan, xanthan gum, gellan gum, gum arabic, and carboxymethyl cellulose, etc. The fabrication techniques, characterizations, as well as in vitro and in vivo evaluations of each PEC delivery system are discussed in detail. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Geographic information system-coupling sediment delivery distributed modeling based on observed data.

    Science.gov (United States)

    Lee, S E; Kang, S H

    2014-01-01

    Spatially distributed sediment delivery (SEDD) models are of great interest in estimating the expected effect of changes on soil erosion and sediment yield. However, they can only be applied if the model can be calibrated using observed data. This paper presents a geographic information system (GIS)-based method to calculate the sediment discharge from basins to coastal areas. For this, an SEDD model, with a sediment rating curve method based on observed data, is proposed and validated. The model proposed here has been developed using the combined application of the revised universal soil loss equation (RUSLE) and a spatially distributed sediment delivery ratio, within Model Builder of ArcGIS's software. The model focuses on spatial variability and is useful for estimating the spatial patterns of soil loss and sediment discharge. The model consists of two modules, a soil erosion prediction component and a sediment delivery model. The integrated approach allows for relatively practical and cost-effective estimation of spatially distributed soil erosion and sediment delivery, for gauged or ungauged basins. This paper provides the first attempt at estimating sediment delivery ratio based on observed data in the monsoon region of Korea.

  6. Expanding delivery system research in public health settings: lessons from practice-based research networks.

    Science.gov (United States)

    Mays, Glen P; Hogg, Rachel A

    2012-11-01

    Delivery system research to identify how best to organize, finance, and implement health improvement strategies has focused heavily on clinical practice settings, with relatively little attention paid to public health settings-where research is made more difficult by wide heterogeneity in settings and limited sources of existing data and measures. This study examines the approaches used by public health practice-based research networks (PBRNs) to expand delivery system research and evidence-based practice in public health settings. Practice-based research networks employ quasi-experimental research designs, natural experiments, and mixed-method analytic techniques to evaluate how community partnerships, economic shocks, and policy changes impact delivery processes in public health settings. In addition, network analysis methods are used to assess patterns of interaction between practitioners and researchers within PBRNs to produce and apply research findings. Findings from individual PBRN studies elucidate the roles of information exchange, community resources, and leadership and decision-making structures in shaping implementation outcomes in public health delivery. Network analysis of PBRNs reveals broad engagement of both practitioners and researchers in scientific inquiry, with practitioners in the periphery of these networks reporting particularly large benefits from research participation. Public health PBRNs provide effective mechanisms for implementing delivery system research and engaging practitioners in the process. These networks also hold promise for accelerating the translation and application of research findings into public health settings.

  7. Optimization of a Der p 2-based prophylactic DNA vaccine against house dust mite allergy.

    Science.gov (United States)

    Pulsawat, Pinya; Pitakpolrat, Patrawadee; Prompetchara, Eakachai; Kaewamatawong, Theerayuth; Techakriengkrai, Navapon; Sirivichayakul, Sunee; Buranapraditkun, Supranee; Hannaman, Drew; Ruxrungtham, Kiat; Jacquet, Alain

    2013-03-01

    DNA vaccines encoding allergens are promising immunotherapeutics to prevent or to treat allergy through induction of allergen-specific Th1 responses. Despite anti-allergy effects observed in small rodents, DNA-based vaccines are weak immunogens in primates and humans and particularly when administered by conventional injection. The goal of the present study was to improve the immunogenicity of a prophylactic vaccine encoding the major house dust mite allergen Der p 2. In this context, we evaluated the influence of different DNA backbones including notably intron and CpG enriched sequence, the DNA dose, the in vivo delivery by electroporation as well as the heterologous prime boost regimen on the vaccine efficiency. We found that a minimal allergen expression level threshold must be reached to induce the production of specific antibodies but beyond this limit, the intensity of the immune response was independent on the DNA dose and allergen expression. The in vivo DNA delivery by electroporation drastically enhanced the production of specific antibodies but not the IFNg secretion. Vaccination of naïve mice with DNA encoding Der p 2 delivered by electroporation even at very low dose (2μg) prevented the development of house dust mite allergy through Th1-skewed immune response characterized by the drastic reduction of allergen-specific IgE, IL-5 and lung inflammation together with the induction of strong specific IgG2a titers and IFNg secretion. CpG cassette in the DNA backbone does not play a critical role in the efficient prophylaxis. Finally, comparable protective immune responses were observed when using heterologous DNA prime/protein boost or homologous DNA prime/boost. Taken together, these data suggest that the potent Th1 response induced by DNA-based vaccine encoding allergens through electroporation provides the rationale for the evaluation of DNA encoding Der p 2 into HDM allergy clinical trials. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Advances of paclitaxel formulations based on nanosystem delivery technology.

    Science.gov (United States)

    Luo, C; Wang, Y; Chen, Q; Han, X; Liu, X; Sun, J; He, Z

    2012-05-01

    In this paper, an overview of recent advances of paclitaxel formulations based on nanosystems is provided. Paclitaxel is very effective in the treatment of various cancers especially ovarian and breast cancer, but it demonstrates poor aqueous solubility, which results in the difficulty challenging the development of paclitaxel parenteral formulations, so its clinical application is greatly restricted. The conventional paclitaxel formulation uses Cremophor EL and ethanol to solubilize paclitaxel, which could cause severe side effects. Nanotechnology has been widely exploited in the field of antitumor research, and paclitaxel is no exception. In recent decades, a series of novel formulations of paclitaxel based on nanotechnology have been developed, including albumin-bound paclitaxel, polymeric micelle-formulated paclitaxel, polymer-paclitaxel conjugates, liposome encapsulated paclitaxel etc. The common advantage shared with these novel injectable formulations is that they are developed based on nanotechnology and Cremophor EL-free. In addition, these nanoformulations can significantly reduce toxicities of paclitaxel and greatly promote its antitumor efficiency.

  9. A Cassette Based System for Hydrogen Storage and Delivery

    Energy Technology Data Exchange (ETDEWEB)

    Britton Wayne E.

    2006-11-29

    A hydrogen storage system is described and evaluated. This is based upon a cassette, that is a container for managing hydrogen storage materials. The container is designed to be safe, modular, adaptable to different chemistries, inexpensive, and transportable. A second module receives the cassette and provides the necessary infrastructure to deliver hydrogen from the cassette according to enduser requirements. The modular concept has a number of advantages over approaches that are all in one stand alone systems. The advantages of a cassette based system are discussed, along with results from model and laboratory testing.

  10. Formulation and evaluation of lidocaine base ethosomes for transdermal delivery.

    Science.gov (United States)

    Zhu, Xiaoliang; Li, Fuli; Peng, Xuebiao; Zeng, Kang

    2013-08-01

    Although transdermal preparations of local anesthetics have been used to reduce pain caused by skin surgery, these preparations cannot effectively penetrate through the epidermis because of the barrier formed by the stratum corneum and the thick epidermis. Ethosomes can effectively transport drugs across the skin because of their thermodynamic stability, small size, high encapsulation efficiency, and percutaneous penetration. We evaluated lidocaine base ethosomes by measuring their loading efficiency, encapsulation efficiency, thermodynamic stability, and percutaneous penetration capability in vitro, and their effectiveness and cutaneous irritation in vivo. Lidocaine base ethosomes were prepared using the injection-sonication-filter method. Size, loading efficiency, encapsulation efficiency, and stability were evaluated using a Zetasizer and high performance liquid chromatography. Formulation was determined by measuring the maximum encapsulation efficiency in the orthogonal test. Percutaneous penetration efficiency in vitro was analyzed using a Franz-type diffusion cell experiment. In vivo effectiveness was analyzed using the pinprick test. Cutaneous irritancy tests were performed on white guinea pigs, followed by histopathologic analysis. The results were compared with lidocaine liposomes as well as lidocaine delivered in a hydroethanolic solution. Lidocaine base ethosomes composed of 5% (w/w) egg phosphatidyl choline, 35% (w/w) ethanol, 0.2% (w/w) cholesterol, 5% (w/w) lidocaine base, and ultrapure water had a mean maximum encapsulation of 51% ± 4%, a mean particle size of 31 ± 3 nm, and a mean loading efficiency of 95.0% ± 0.1%. The encapsulation efficiency of lidocaine base ethosomes remained stable for 60 days at 25°C ± 1°C (95% confidence interval [CI], -1.12% to 1.34%; P = 0.833). The transdermal flux of lidocaine base differed significantly for the 3 preparations (F = 120, P ethosomes than from liposomes (95% corrected CI, 1129-1818 µg/(cm(2)·h); P

  11. Low-Amplitude Ultrasound Enhances Hydrodynamic-Based Gene Delivery to Rat Kidney

    Science.gov (United States)

    Xing, Yifei; Pua, Eric C.; Lu, Xiaochun; Zhong, Pei

    2009-01-01

    The synergistic combination of hydrodynamics-based gene delivery and ultrasound was investigated to achieve improved gene transfer to the kidney. Plasmids encoding firefly luciferase and Erythropoietin (EPO) gene were delivered into the left kidney of rats by single or combinative application of renal vein hydrodynamic injection and ultrasound treatment with or without the addition of ultrasound contrast agents (UCA). Ultrasound exposure was found to enhance the efficiency of hydrodynamics-based gene delivery for both luciferase and EPO expression. An ultrasound exposure intensity of 2 W/cm2 at 10% duty cycle for 15 min., produced a maximal gene expression 4.5 times higher than hydrodynamic delivery alone. Duration, location, and tissue-specificity of gene expression were not changed by ultrasound exposure. Application of UCA reduced the intensity and exposure duration of ultrasound treatment needed for optimal expression. Appropriate application of ultrasound and UCA did not alter histological structure or impair physiological function of the treated kidney. PMID:19523454

  12. PEG-PCL-based nanomedicines: A biodegradable drug delivery system and its application.

    Science.gov (United States)

    Grossen, Philip; Witzigmann, Dominik; Sieber, Sandro; Huwyler, Jörg

    2017-08-28

    The lack of efficient therapeutic options for many severe disorders including cancer spurs demand for improved drug delivery technologies. Nanoscale drug delivery systems based on poly(ethylene glycol)-poly(ε-caprolactone) copolymers (PEG-PCL) represent a strategy to implement therapies with enhanced drug accumulation at the site of action and decreased off-target effects. In this review, we discuss state-of-the-art nanomedicines based on PEG-PCL that have been investigated in a preclinical setting. We summarize the various synthesis routes and different preparation methods used for the production of PEG-PCL nanoparticles. Additionally, we review physico-chemical properties including biodegradability, biocompatibility, and drug loading. Finally, we highlight recent therapeutic applications investigated in vitro and in vivo using advanced systems such as triggered release, multi-component therapies, theranostics, or gene delivery systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Drug delivery systems based on nucleic acid nanostructures

    NARCIS (Netherlands)

    de Vries, Jan Willem; Zhang, Feng; Herrmann, Andreas

    2013-01-01

    The field of DNA nanotechnology has progressed rapidly in recent years and hence a large variety of 1D-, 2D-and 3D DNA nanostructures with various sizes, geometries and shapes is readily accessible. DNA-based nanoobjects are fabricated by straight forward design and self-assembly processes allowing

  14. Monolithic natural gas storage delivery system based on sorbents

    Energy Technology Data Exchange (ETDEWEB)

    Hornbostel, Marc; Krishnan, Gopala N.; Sanjurjo, Angel

    2016-09-27

    The invention provides methods for producing a strong, light, sorbent-based storage/dispenser system for gases and fuels. The system comprises a porous monolithic material with an adherent strong impervious skin that is capable of storing a gas under pressure in a safe and usable manner.

  15. Stem cell-based therapy : Improving myocardial cell delivery

    NARCIS (Netherlands)

    Feyen, Dries A M|info:eu-repo/dai/nl/413647838; Gaetani, RG; Doevendans, Pieter A.|info:eu-repo/dai/nl/164248366; Sluijter, Joost P G|info:eu-repo/dai/nl/273307908

    2016-01-01

    Stem cell-based therapies form an exciting new class of medicine that attempt to provide the body with the building blocks required for the reconstruction of damaged organs. However, delivering cells to the correct location, while preserving their integrity and functional properties, is a complex

  16. Crystals and crystallization in oil-in-water emulsions: implications for emulsion-based delivery systems.

    Science.gov (United States)

    McClements, David Julian

    2012-06-15

    Many bioactive components intended for oral ingestion (pharmaceuticals and nutraceuticals) are hydrophobic molecules with low water-solubilities and high melting points, which poses considerable challenges to the formulation of oral delivery systems. Oil-in-water emulsions are often suitable vehicles for the encapsulation and delivery of this type of bioactive component. The bioactive component is usually dissolved in a carrier lipid phase by either dilution and/or heating prior to homogenization, and then the carrier lipid and water phases are homogenized to form an emulsion consisting of small oil droplets dispersed in water. The successful development of this kind of emulsion-based delivery system depends on a good understanding of the influence of crystals on the formation, stability, and properties of emulsions. This review article addresses the physicochemical phenomena associated with the encapsulation, retention, crystallization, release, and absorption of hydrophobic bioactive components within emulsions. This knowledge will be useful for the rational formulation of effective emulsion-based delivery systems for oral delivery of crystalline hydrophobic bioactive components in the food, health care, and pharmaceutical industries. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Efficient genetic engineering of human intestinal organoids using electroporation.

    Science.gov (United States)

    Fujii, Masayuki; Matano, Mami; Nanki, Kosaku; Sato, Toshiro

    2015-10-01

    Gene modification in untransformed human intestinal cells is an attractive approach for studying gene function in intestinal diseases. However, because of the lack of practical tools, such studies have largely depended upon surrogates, such as gene-engineered mice or immortalized human cell lines. By taking advantage of the recently developed intestinal organoid culture method, we developed a methodology for modulating genes of interest in untransformed human colonic organoids via electroporation of gene vectors. Here we describe a detailed protocol for the generation of intestinal organoids by culture with essential growth factors in a basement membrane matrix. We also describe how to stably integrate genes via the piggyBac transposon, as well as precise genome editing using the CRISPR-Cas9 system. Beginning with crypt isolation from a human colon sample, genetically modified organoids can be obtained in 3 weeks.

  18. Lightning-triggered electroporation and electrofusion as possible contributors to natural HGT among prokaryotes

    CERN Document Server

    Kotnik, Tadej

    2012-01-01

    Phylogenetic studies show that horizontal gene transfer (HGT) is a significant contributor to genetic variability of prokaryotes, and was perhaps even more abundant during early evolution. Hitherto, research of natural HGT has mainly focused on three mechanisms: conjugation, natural competence, and viral transduction. This paper discusses the feasibility of a fourth such mechanism - cell electroporation and/or electrofusion triggered by atmospheric electrostatic discharges (lightnings). A description of electroporation as a phenomenon is followed by a review of experimental evidence that electroporation of prokaryotes in aqueous environments can result in release of non-denatured DNA, as well as uptake of DNA from the surroundings and transformation. Similarly, a description of electrofusion is followed by a review of experiments showing that prokaryotes devoid of cell wall can electrofuse into hybrids expressing the genes of their both precursors. Under sufficiently fine-tuned conditions, electroporation and...

  19. Theoretical analysis of AC electric field transmission into biological tissue through frozen saline for electroporation.

    Science.gov (United States)

    Xiao, Chunyan; Rubinsky, Boris

    2014-12-01

    An analytical model was used to explore the feasibility of sinusoidal electric field transmission across a frozen saline layer into biological tissue. The study is relevant to electroporation and permeabilization of the cell membrane by electric fields. The concept was analyzed for frequencies in the range of conventional electroporation frequencies and electric field intensity. Theoretical analysis for a variety of tissues show that the transmission of electroporation type electric fields through a layer of frozen saline into tissue is feasible and the behavior of this composite system depends on tissue type, frozen domain temperature, and frequency. Freezing could become a valuable method for adherence of electroporation electrodes to moving tissue surfaces, such as the heart in the treatment of atrial fibrillation or blood vessels for the treatment of restenosis. © 2014 Wiley Periodicals, Inc.

  20. Electroporation enhances mitomycin C cytotoxicity on T24 bladder cancer cell line

    DEFF Research Database (Denmark)

    Vasquez, Juan Luis; Gehl, Julie; Hermann, Gregers G

    2012-01-01

    Intravesical mitomycin instillation combined with electric pulses is being used experimentally for the treatment of T1 bladder tumors, in patients unfit for surgery. Electroporation may enhance the uptake of chemotherapeutics by permeabilization of cell membranes. We investigated if electroporation...... improves the cytotoxicity of mitomycin. In two cell lines, T24 (bladder cancer cell line) and DC3F (Chinese hamster fibroblast), exposure to different concentrations of mitomycin (0.01-2000μM) was tested with and without electroporation (6 pulses of 1kV/cm, duration: 99μs, frequency: 1Hz). Cell viability...... was assessed by colorimetric assay (MTT). For both cell lines, mitomycin's IC_50 was approximately 1000μM in both pulsed and unpulsed cells. On T24 cells, electroporation and mitomycin caused (relative reduction) RR of survival of: 25%, 31% and 29%, by concentrations 0μM, 500μM and 1000μM respectively. For DC3...

  1. Social services delivery through community-based projects

    OpenAIRE

    Dinah McLeod; Maurizia Tovo

    2001-01-01

    The World Bank is financing an increasing number of community-based social services projects. The objective of this paper is to review and categorize the extent, scope and mechanisms of these projects in the current Bank portfolio, and to identify good practices and potential pitfalls. The authors identify 99 projects that finance at least $1.6 billion in social services. While most of the projects surveyed deliver"traditional"services such as nutrition, maternal and child care, and literacy,...

  2. CHO-S antibody titers >1 gram/liter using flow electroporation-mediated transient gene expression followed by rapid migration to high-yield stable cell lines.

    Science.gov (United States)

    Steger, Krista; Brady, James; Wang, Weili; Duskin, Meg; Donato, Karen; Peshwa, Madhusudan

    2015-04-01

    In recent years, researchers have turned to transient gene expression (TGE) as an alternative to CHO stable cell line generation for early-stage antibody development. Despite advances in transfection methods and culture optimization, the majority of CHO-based TGE systems produce insufficient antibody titers for extensive use within biotherapeutic development pipelines. Flow electroporation using the MaxCyte STX Scalable Transfection System is a highly efficient, scalable means of CHO-based TGE for gram-level production of antibodies without the need for specialized expression vectors or genetically engineered CHO cell lines. CHO cell flow electroporation is easily scaled from milligram to multigram quantities without protocol reoptimization while maintaining transfection performance and antibody productivity. In this article, data are presented that demonstrate the reproducibility, scalability, and antibody production capabilities of CHO-based TGE using the MaxCyte STX. Data show optimization of posttransfection parameters such as cell density, media composition, and feed strategy that result in secreted antibody titers >1 g/L and production of multiple grams of antibody within 2 weeks of a single CHO-S cell transfection. In addition, data are presented to demonstrate the application of scalable electroporation for the rapid generation of high-yield stable CHO cell lines to bridge the gap between early- and late-stage antibody development activities. © 2014 Society for Laboratory Automation and Screening.

  3. Electroporation in the Regenerating Tail of the Xenopus Tadpole

    Science.gov (United States)

    Mochii, Makoto; Taniguchi, Yuka

    Xenopus laevis is a model system widely used to investigate embryogenesis, metamorphosis, and regeneration. The tail of the Xenopus tadpole is very useful in analyzing the molecular mechanisms underlying appendage regeneration (Slack et al., 2004; Mochii et al., 2007; Slack et al., 2008). It is transparent and suitable for whole-mount observation at the cellular level. The tail regenerates within 2 weeks of amputation. The conventional injection of blastomeres with mRNA, DNA, or antisense oligonucleotides is a powerful tool with which to study genetic mechanisms in early embryos, but it is not effective in late embryos or larvae. A transgenic approach has been used to analyze tail regeneration (Beck et al., 2003, 2006), but its success is largely dependent on the activity of the promoter used. There are limited numbers of promoters available that precisely regulate gene expression spatially and/or temporally. In vivo electroporation is an alternative method that can be used to manipulate gene expression in late embryos and larvae. The introduction of DNA or RNA into the cells of neurula and tailbud embryos has been reported (Eide et al., 2000; Sasagawa et al., 2002; Falk et al., 2007). Targeting larval tissues with in vivo electroporation also has been used to investigate neural networks, metamorphosis, and regeneration (Haas et al., 2001, 2002; Nakajima and Yaoita, 2003; Javaherian and Cline, 2005; Bestman et al., 2006; Boorse et al., 2006; Lin et al., 2007; Mochii et al., 2007). In this chapter, we report a procedure to introduce DNA into the tissues of the tadpole tail.

  4. Noninvasive measurement of transdermal drug delivery by impedance spectroscopy.

    Science.gov (United States)

    Arpaia, Pasquale; Cesaro, Umberto; Moccaldi, Nicola

    2017-03-24

    The effectiveness in transdermal delivery of skin permeation strategies (e.g., chemical enhancers, vesicular carrier systems, sonophoresis, iontophoresis, and electroporation) is poorly investigated outside of laboratory. In therapeutic application, the lack of recognized techniques for measuring the actually-released drug affects the scientific concept itself of dosage for topically- and transdermally-delivered drugs. Here we prove the suitability of impedance measurement for assessing the amount of drug penetrated into the skin after transdermal delivery. In particular, the measured amount of drug depends linearly on the impedance magnitude variation normalized to the pre-treated value. Three experimental campaigns, based on the electrical analysis of the biological tissue behavior due to the drug delivery, are reported: (i) laboratory emulation on eggplants, (ii) ex-vivo tests on pig ears, and finally (iii) in-vivo tests on human volunteers. Results point out that the amount of delivered drug can be assessed by reasonable metrological performance through a unique measurement of the impedance magnitude at one single frequency. In particular, in-vivo results point out sensitivity of 23 ml-1, repeatability of 0.3%, non-linearity of 3.3%, and accuracy of 5.7%. Finally, the measurement resolution of 0.20 ml is compatible with clinical administration standards.

  5. Changes in optical properties of electroporated cells as revealed by digital holographic microscopy

    OpenAIRE

    Calin, Violeta L.; Mihailescu, Mona; Mihale, Nicolae; Baluta, Alexandra V.; Kovacs, Eugenia; Savopol, Tudor; Moisescu, Mihaela G.

    2017-01-01

    Changes in optical and shape-related characteristics of B16F10 cells after electroporation were investigated using digital holographic microscopy (DHM). Bipolar rectangular pulses specific for electrochemotherapy were used. Electroporation was performed in an ?off-axis? DHM set-up without using exogenous markers. Two types of cell parameters were monitored seconds and minutes after pulse train application: parameters addressing a specifically defined area of the cell (refractive index and cel...

  6. High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA.

    OpenAIRE

    Miller, J F; Dower, W J; Tompkins, L S

    1988-01-01

    Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of high-voltage discharges to bacterial cells permits genetic transformation. Our method involves exposure of...

  7. Live delivery outcome after tubal sterilization reversal: a population-based study.

    Science.gov (United States)

    Malacova, Eva; Kemp-Casey, Anna; Bremner, Alexandra; Hart, Roger; Stewart, Louise Maree; Preen, David Brian

    2015-10-01

    To determine the cumulative incidence of live delivery in women who underwent reversal of tubal sterilization. Population-based retrospective cohort study. Hospitals in Western Australia. All women aged 20-44 years, with a history of hospital admission for tubal sterilization, who subsequently underwent reversal of sterilization during the period 1985 to 2009 in Western Australia (n = 1,898). Data regarding reversal of sterilization and prior tubal sterilization were extracted from routinely collected administrative hospital separation records, until commencement of IVF treatment. First live-delivery rates. There were 969 first live deliveries observed during the study period. The overall cumulative live-delivery rate was 20% (95% confidence interval [CI] 18-23) within the first year after reversal, 40% (95% CI 38-42) at 2 years, 51% (95% CI 48-53) at 5 years, and 52% (95% CI 50-55) at 10 years. The 5-year cumulative live-delivery rate was significantly lower in women who were aged 40-44 years (26%) compared with younger women (aged 20-29, 30-34, and 35-39 years) (50%, 56%, and 51%, respectively). Women undergoing reversal of sterilization before they reach age 40 years have at least a 50% chance of delivering a live baby within the next 5 years. Up to that age, there is no significant difference in live deliveries. The live-delivery rate halves after the age of 40 years. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Mode of delivery and the probability of subsequent childbearing: a population-based register study.

    Science.gov (United States)

    Elvander, C; Dahlberg, J; Andersson, G; Cnattingius, S

    2015-11-01

    To investigate the relationship between mode of first delivery and probability of subsequent childbearing. Population-based study. Nationwide study in Sweden. A cohort of 771 690 women who delivered their first singleton infant in Sweden between 1992 and 2010. Using Cox's proportional-hazards regression models, risks of subsequent childbearing were compared across four modes of delivery. Hazard ratios (HRs) were calculated, using 95% confidence intervals (95% CIs). Probability of having a second and third child; interpregnancy interval. Compared with women who had a spontaneous vaginal first delivery, women who delivered by vacuum extraction were less likely to have a second pregnancy (HR 0.96, 95% CI 0.95-0.97), and the probabilities of a second childbirth were substantially lower among women with a previous emergency caesarean section (HR 0.85, 95% CI 0.84-0.86) or an elective caesarean section (HR 0.82, 95% CI 0.80-0.83). There were no clinically important differences in the median time between first and second pregnancy by mode of first delivery. Compared with women younger than 30 years of age, older women were more negatively affected by a vacuum extraction with respect to the probability of having a second child. A primary vacuum extraction decreased the probability of having a third child by 4%, but having two consecutive vacuum extraction deliveries did not further alter the probability. A first delivery by vacuum extraction does not reduce the probability of subsequent childbearing to the same extent as a first delivery by emergency or elective caesarean section. © 2014 Royal College of Obstetricians and Gynaecologists.

  9. Polysaccharide-based micro/nanocarriers for oral colon-targeted drug delivery.

    Science.gov (United States)

    Zhang, Lin; Sang, Yuan; Feng, Jing; Li, Zhaoming; Zhao, Aili

    2016-08-01

    Oral colon-targeted drug delivery has attracted many researchers because of its distinct advantages of increasing the bioavailability of the drug at the target site and reducing the side effects. Polysaccharides that are precisely activated by the physiological environment of the colon hold greater promise for colon targeting. Considerable research efforts have been directed towards developing polysaccharide-based micro/nanocarriers. Types of polysaccharides for colon targeting and in vitro/in vivo assessments of polysaccharide-based carriers for oral colon-targeted drug delivery are summarised. Polysaccharide-based microspheres have gained increased importance not just for the delivery of the drugs for the treatment of local diseases associated with the colon (colon cancer, inflammatory bowel disease (IBD), amoebiasis and irritable bowel syndrome (IBS)), but also for it's potential for the delivery of anti-rheumatoid arthritis and anti-chronic stable angina drugs. Besides, Polysaccharide-based micro/nanocarriers such as microbeads, microcapsules, microparticles, nanoparticles, nanogels and nanospheres are also introduced in this review.

  10. The Effect of Educational Intervention on Selection of Delivery Method Based on Health Belief Model.

    Directory of Open Access Journals (Sweden)

    Mohammad Hossein Baghianimoghadam

    2014-07-01

    Discussion- In this study educational intervention based on health belief model increased the awareness of pregnant women, However ,it has not been effective on their performance. Because many factors other than knowledge are involved in the choice of delivery method, It is proposed to enhance the efficiency of this model simultaneously different patterns that can be used effectively on other factors.

  11. Designing polymers with sugar-based advantages for bioactive delivery applications.

    Science.gov (United States)

    Zhang, Yingyue; Chan, Jennifer W; Moretti, Alysha; Uhrich, Kathryn E

    2015-12-10

    Sugar-based polymers have been extensively explored as a means to increase drug delivery systems' biocompatibility and biodegradation. Here,we review he use of sugar-based polymers for drug delivery applications, with a particular focus on the utility of the sugar component(s) to provide benefits for drug targeting and stimuli responsive systems. Specifically, numerous synthetic methods have been developed to reliably modify naturally-occurring polysaccharides, conjugate sugar moieties to synthetic polymer scaffolds to generate glycopolymers, and utilize sugars as a multifunctional building block to develop sugar-linked polymers. The design of sugar-based polymer systems has tremendous implications on both the physiological and biological properties imparted by the saccharide units and are unique from synthetic polymers. These features include the ability of glycopolymers to preferentially target various cell types and tissues through receptor interactions, exhibit bioadhesion for prolonged residence time, and be rapidly recognized and internalized by cancer cells. Also discussed are the distinct stimuli-sensitive properties of saccharide-modified polymers to mediate drug release under desired conditions. Saccharide-based systems with inherent pH- and temperature-sensitive properties, as well as enzyme-cleavable polysaccharides for targeted bioactive delivery, are covered. Overall, this work emphasizes inherent benefits of sugar-containing polymer systems for bioactive delivery.

  12. Spontaneous preterm delivery among primiparous women at low risk in Denmark: a population based study

    DEFF Research Database (Denmark)

    Langhoff-Roos, Jens; Kesmodel, Ulrik; Jacobsson, Bo

    2006-01-01

    can be compared internationally. DESIGN: Population based study. PARTICIPANTS: 99.8% of all deliveries in Denmark, 1995-2004. MAIN OUTCOME MEASURES: Proportion of babies born at less than 37 weeks' completed gestation for each year in the overall population and in a standard population at low risk...

  13. Generic project definitions for improvement of health care delivery: A case-base approach

    NARCIS (Netherlands)

    Niemeijer, G.C.; Does, R.J.M.M.; de Mast, J.; Trip, A.; van den Heuvel, J.

    2011-01-01

    Background: The purpose of this article is to create actionable knowledge, making the definition of process improvement projects in health care delivery more effective. Methods: This study is a retrospective analysis of process improvement projects in hospitals, facilitating a case-based reasoning

  14. Polysaccharide-based aerogel microspheres for oral drug delivery.

    Science.gov (United States)

    García-González, C A; Jin, M; Gerth, J; Alvarez-Lorenzo, C; Smirnova, I

    2015-03-06

    Polysaccharide-based aerogels in the form of microspheres were investigated as carriers of poorly water soluble drugs for oral administration. These bio-based carriers may combine the biocompatibility of polysaccharides and the enhanced drug loading capacity of dry aerogels. Aerogel microspheres from starch, pectin and alginate were loaded with ketoprofen (anti-inflammatory drug) and benzoic acid (used in the management of urea cycle disorders) via supercritical CO2-assisted adsorption. Amount of drug loaded depended on the aerogel matrix structure and composition and reached values up to 1.0×10(-3) and 1.7×10(-3) g/m(2) for ketoprofen and benzoic acid in starch microspheres. After impregnation, drugs were in the amorphous state in the aerogel microspheres. Release behavior was evaluated in different pH media (pH 1.2 and 6.8). Controlled drug release from pectin and alginate aerogel microspheres fitted Gallagher-Corrigan release model (R(2)>0.99 in both cases), with different relative contribution of erosion and diffusion mechanisms depending on the matrix composition. Release from starch aerogel microspheres was driven by dissolution, fitting the first-order kinetics due to the rigid starch aerogel structure, and showed different release rate constant (k1) depending on the drug (0.075 and 0.160 min(-1) for ketoprofen and benzoic acid, respectively). Overall, the results point out the possibilities of tuning drug loading and release by carefully choosing the polysaccharide used to prepare the aerogels. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Biodegradable Oxamide-Phenylene-Based Mesoporous Organosilica Nanoparticles with Unprecedented Drug Payloads for Delivery in Cells

    KAUST Repository

    Croissant, Jonas

    2016-06-03

    We describe biodegradable mesoporous hybrid NPs in the presence of proteins, and its application for drug delivery. We synthesized oxamide-phenylene-based mesoporous organosilica nanoparticles (MON) in the absence of silica source which had a remarkably high organic content with a high surface area. Oxamide functions provided biodegradability in the presence of trypsin model proteins. MON displayed exceptionally high payloads of hydrophilic and hydrophobic drugs (up to 84 wt%), and a unique zero premature leakage without the pore capping, unlike mesoporous silica. MON were biocompatible and internalized into cancer cells for drug delivery.

  16. Cell-penetrating peptide-based intelligent liposomal systems for enhanced drug delivery.

    Science.gov (United States)

    Gao, Huile; Zhang, Qianyu; Yu, Zhiqiang; He, Qin

    2014-01-01

    Liposomes are widely used as drug delivery systems and several liposome-based nanomedicines have been approved for clinical use. Cell penetrating peptides (CPPs) have been decorated onto nanoparticulated vesicle such as liposomes to further improve the intracellular delivery efficiency. However, the poor selectivity of CPPs hindered their application, especially in the in vivo application. To resolve this issue, several strategies have been developed, including shielding and environment-triggered deshielding of CPPs as well as designing of environment-responsive CPPs and specific- targeting CPPs and last but not least, combination strategy. In this review, the abovementioned strategies were discussed.

  17. Design, fabrication and characterization of drug delivery systems based on lab-on-a-chip technology.

    Science.gov (United States)

    Nguyen, Nam-Trung; Shaegh, Seyed Ali Mousavi; Kashaninejad, Navid; Phan, Dinh-Tuan

    2013-11-01

    Lab-on-a-chip technology is an emerging field evolving from the recent advances of micro- and nanotechnologies. The technology allows the integration of various components into a single microdevice. Microfluidics, the science and engineering of fluid flow in microscale, is the enabling underlying concept for lab-on-a-chip technology. The present paper reviews the design, fabrication and characterization of drug delivery systems based on this amazing technology. The systems are categorized and discussed according to the scales at which the drug is administered. Starting with the fundamentals on scaling laws of mass transfer and basic fabrication techniques, the paper reviews and discusses drug delivery devices for cellular, tissue and organism levels. At the cellular level, a concentration gradient generator integrated with a cell culture platform is the main drug delivery scheme of interest. At the tissue level, the synthesis of smart particles as drug carriers using lab-on-a-chip technology is the main focus of recent developments. At the organism level, microneedles and implantable devices with fluid-handling components are the main drug delivery systems. For drug delivery to a small organism that can fit into a microchip, devices similar to those of cellular level can be used. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Caesarean Delivery and Postpartum Maternal Mortality: A Population-Based Case Control Study in Brazil.

    Science.gov (United States)

    Esteves-Pereira, Ana Paula; Deneux-Tharaux, Catherine; Nakamura-Pereira, Marcos; Saucedo, Monica; Bouvier-Colle, Marie-Hélène; Leal, Maria do Carmo

    2016-01-01

    Cesarean delivery rates continue to increase worldwide and reached 57% in Brazil in 2014. Although the safety of this surgery has improved in the last decades, this trend is a concern because it carries potential risks to women's health and may be a modifiable risk factor of maternal mortality. This paper aims to investigate the risk of postpartum maternal death directly associated with cesarean delivery in comparison to vaginal delivery in Brazil. This was a population-based case-control study performed in eight Brazilian states. To control for indication bias, deaths due to antenatal morbidity were excluded. We included 73 cases of postpartum maternal deaths from 2009-2012. Controls were selected from the Birth in Brazil Study, a 2011 nationwide survey including 9,221 postpartum women. We examined the association of cesarean section and postpartum maternal death by multivariate logistic regression, adjusting for confounders. After controlling for indication bias and confounders, the risk of postpartum maternal death was almost three-fold higher with cesarean than vaginal delivery (OR 2.87, 95% CI 1.63-5.06), mainly due to deaths from postpartum hemorrhage and complications of anesthesia. Cesarean delivery is an independent risk factor of postpartum maternal death. Clinicians and patients should consider this fact in balancing the benefits and risks of the procedure.

  19. Update on Nanotechnology-based Drug Delivery Systems in Cancer Treatment.

    Science.gov (United States)

    Ho, Benjamin N; Pfeffer, Claire M; Singh, Amareshwar T K

    2017-11-01

    The emerging field of nanotechnology meets the demands for innovative approaches in the diagnosis and treatment of cancer. The nanoparticles are biocompatible and biodegradable and are made of a core, a particle that acts as a carrier, and one or more functional groups on the core which target specific sites. Nanotech in drug delivery includes nanodisks, High Density Lipoprotein nanostructures, liposomes, and gold nanoparticles. The fundamental advantages of nanoparticles are: improved delivery of water-insoluble drugs, targeted delivery, co-delivery of two or more drugs for combination therapy, and visualization of the drug delivery site by combining imaging system and a therapeutic drug. One of the potential applications of nanotechnology is in the treatment of cancer. Conventional methods for cancer treatments have included chemotherapy, surgery, or radiation. Early recognition and treatment of cancer with these approaches is still challenging. Innovative technologies are needed to overcome multidrug resistance, and increase drug localization and efficacy. Application of nanotechnology to cancer biology has brought in a new hope for developing treatment strategies on cancer. In this study, we present a review on the recent advances in nanotechnology-based approaches in cancer treatment. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  20. Miconazole Microsponges based topical delivery system for diaper dermatitis

    Directory of Open Access Journals (Sweden)

    Neha Gulati

    Full Text Available Aim: The current investigation was aimed to develop and optimize the microsponges of miconazole nitrate for treatment of diaper dermatitis for enhanced therapeutic effect. Material and Methods: Microsponges were developed by emulsion solvent diffusion technique using 2³ factorial design. Fabricated microsponges were optimized in order to analyze the effects of independent variables on the encapsulation efficiency, particle size, surface topography and in vitro drug release. The optimized formulation was then incorporated into the gel and evaluated. Results: Particle size of all formulations was found to be uniform and scanning electron microscopy (SEM indicates spherical shape and porous nature of microsponges. In vitro drug release studies of all formulations revealed the release rate within the range of 67%±0.09 to 80.6%± 0.68 at the end of 12 hours. On its basis, formulation F8 was selected and incorporated into the gel (CF8 which was evaluated for pH, viscosity, spreadability, in vitro drug diffusion studies, in vitro anti fungal studies and stability studies. Conclusion: The formulated microsponge-based gel of miconazole nitrate would be a capable substitute to traditional treatment for reliable and economical cure of diaper dermatitis.

  1. Encapsulation and delivery of food ingredients using starch based systems.

    Science.gov (United States)

    Zhu, Fan

    2017-08-15

    Functional ingredients can be encapsulated by various wall materials for controlled release in food and digestion systems. Starch, as one of the most abundant natural carbohydrate polymers, is non-allergenic, GRAS, and cheap. There has been increasing interest of using starch in native and modified forms to encapsulate food ingredients such as flavours, lipids, polyphenols, carotenoids, vitamins, enzymes, and probiotics. Starches from various botanical sources in granular or amorphous forms are modified by chemical, physical, and/or enzymatic means to obtain the desired properties for targeted encapsulation. Other wall materials are also employed in combination with starch to facilitate some types of encapsulation. Various methods of crafting the starch-based encapsulation such as electrospinning, spray drying, antisolvent, amylose inclusion complexation, and nano-emulsification are introduced in this mini-review. The physicochemical and structural properties of the particles are described. The encapsulation systems can positively influence the controlled release of food ingredients in food and nutritional applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Carbopol-based gels for nasal delivery of progesterone.

    Science.gov (United States)

    Rathnam, Grace; Narayanan, N; Ilavarasan, R

    2008-01-01

    The purpose of this study was to investigate the nasal absorption of progesterone from carbopol-based nasal gels in rabbits. Progesterone nasal gels were prepared by dispersing carbopol 974 (1%, 1.5%, and 2%) in distilled water followed by addition of progesterone/progesterone-beta cyclodextrin complex dissolved in propylene glycol then neutralization. The potential use of beta cyclodextrin (CD) as nasal absorption enhancer by simple addition, as a physical mixture and as a complex with progesterone was investigated. The absolute bioavailability of progesterone from nasal gels in rabbits was studied by estimating the serum progesterone level by competitive solid-phase enzyme immunoassay in comparison to intravenous injection. The carbopol gel formulations produced a significant increase in bioavailability. CD complex promotes the nasal absorption of progesterone from carbopol gels as compared with gels where the CD is added by simple addition and gels which do not contain CD. This method of addition of CD as an inclusion complex in the gels could be considered as a preferred platform in nasal drug administration.

  3. Minimally invasive intracellular delivery based on electrokinetic forces combined with vibration-assisted cell membrane perforation

    Science.gov (United States)

    Shibata, Takayuki; Ozawa, Tatsuya; Ito, Yasuharu; Yamamoto, Keita; Nagai, Moeto

    2017-01-01

    To provide an effective platform for the fundamental analysis of cellular mechanisms and the regulation of cellular functions, we developed a unique method of minimally invasive intracellular delivery. Using this method, we successfully demonstrated the delivery of DNA molecules into living HeLa cells via a glass micropipette based on DC-biased AC-driven electrokinetic forces with much better controllability than that of the pressure-driven flow method. We also proposed a vibration-assisted insertion method for penetrating the cell membrane to reduce cell damage. Preliminary insertion tests revealed that application of mechanical oscillation can reduce the deformation of cells due to increases in their viscous resistance, resulting in a high probability of cell membrane perforation and cell viability. Moreover, to overcome the intrinsic low throughput of intracellular delivery with a single glass micropipette, we developed a fabrication process involving an array of stepped hollow silicon dioxide (SiO2) nanoneedles with well-defined tips.

  4. Clay-based drug-delivery systems: what does the future hold?

    Science.gov (United States)

    Giuseppe; Riela, Serena; Fakhrullin, Rawil F

    2017-07-01

    Clays for drug delivery have been used from ancient time due to the large availability of clay minerals and their unprecedented properties. The empirical use of nanoclays from the past is converted in a stimulating scientific task aimed at building up nanoarchitectonic vehicles for drug delivery in a targeted and stimuli-responsive fashion. Here the historical aspects are discussed; next the modern examples of applications of different clay-based materials are discussed. A special focus is given to halloysite clay nanotubes, which are an emerging and very promising nanomaterial for drug-delivery purposes due to its special morphology and unique chemical properties. Advantages and limitations of these natural nanomaterials are critically discussed pointing out the future perspectives and directions for further research.

  5. Breech at term--mode of delivery? A register-based study

    DEFF Research Database (Denmark)

    Krebs, L; Langhoff-Roos, J; Weber, Tom

    1995-01-01

    . Register-based cohort study of all (n = 15718) singleton term breech deliveries of non-malformed infants in Denmark 1982-1990. Process and outcome measures: mode of delivery, gestational age, birth weight, congenital malformations, intrapartum death, Apgar scores and early neonatal death. RESULTS. A total...... of 3247 (20.7%) term infants were delivered vaginally, 7106 (45.3%) by elective and 5356 (34.1%) by emergency cesarean section. Infants delivered vaginally and by emergency cesarean section had significantly higher rates of mortality (intrapartum and early neonatal death) and morbidity (low Apgar scores...... that vaginal delivery is associated with increased mortality and morbidity. However, validation of data and additional information from the medical records are needed before a recommendation of whether selection of parturients, structure of perinatal care or professional skills need to be improved, or all...

  6. A review of drug delivery systems based on nanotechnology and green chemistry: green nanomedicine.

    Science.gov (United States)

    Jahangirian, Hossein; Lemraski, Ensieh Ghasemian; Webster, Thomas J; Rafiee-Moghaddam, Roshanak; Abdollahi, Yadollah

    2017-01-01

    This review discusses the impact of green and environmentally safe chemistry on the field of nanotechnology-driven drug delivery in a new field termed "green nanomedicine". Studies have shown that among many examples of green nanotechnology-driven drug delivery systems, those receiving the greatest amount of attention include nanometal particles, polymers, and biological materials. Furthermore, green nanodrug delivery systems based on environmentally safe chemical reactions or using natural biomaterials (such as plant extracts and microorganisms) are now producing innovative materials revolutionizing the field. In this review, the use of green chemistry design, synthesis, and application principles and eco-friendly synthesis techniques with low side effects are discussed. The review ends with a description of key future efforts that must ensue for this field to continue to grow.

  7. Costs of a Hospital-Based, Ready-To-Use Syringe Delivery Programme

    DEFF Research Database (Denmark)

    Risør, Bettina Wulff; Vand, S; Lisby, M

    2017-01-01

    . Local unit costs were converted to 2013-€ to estimate the incremental costs. Results: The analysis showed that the ready-to-use programme was more costly than the conventional delivery programme. The annual incremental cost for the day surgery department was estimated at €70,469 (an increase of 105......Objective: The risk of errors in the medication administration process is high. Applications of pre lled syringes may improve patient safety but could be more costly. The objective of this study was to assess the additional costs of a ready-to-use syringe delivery programme in comparison...... with a conventional delivery programme at day surgery and endoscopy departments at a large university hospital. Methods: The cost analysis used the hospital perspective and developed an “activity-based costing” model to assess the costs of medicine- handling activities. The model was calibrated with six-month data...

  8. A targeted drug delivery system based on dopamine functionalized nano graphene oxide

    Science.gov (United States)

    Masoudipour, Elham; Kashanian, Soheila; Maleki, Nasim

    2017-01-01

    The cellular targeting property of a biocompatible drug delivery system can widely increase the therapeutic effect against various diseases. Here, we report a dopamine conjugated nano graphene oxide (DA-nGO) carrier for cellular delivery of the anticancer drug, Methotrexate (MTX) into DA receptor positive human breast adenocarcinoma cell line. The material was characterized using scanning electron microscopy, atomic force microscopy, Fourier transform infrared spectroscopy and UV-vis spectroscopy. Furthermore, the antineoplastic action of MTX loaded DA-nGO against DA receptor positive and negative cell lines were explored. The results presented in this article demonstrated that the application of DA functionalized GO as a targeting drug carrier can improve the drug delivery efficacy for DA receptor positive cancer cell lines and promise future designing of carrier conjugates based on it.

  9. Concordance-based adherence support service delivery: consumer perspectives.

    Science.gov (United States)

    Du Pasquier, Sophie; Aslani, Parisa

    2008-12-01

    pressures, financial constraints, the gap of competence and power between patients and doctors. Pharmacists should consider consumers' needs for information and establishing concordance, as well as their expectations of the pharmacy profession, in delivering concordance based adherence support services. Given participants' high demand for medicine information, an opportunity might exist for pharmacists to influence consumers' expectations by offering information which is tailored towards their needs.

  10. Planning and delivery comparison of six linac-based stereotactic radiosurgery techniques

    Science.gov (United States)

    Thakur, Varun Singh

    This work presents planning and delivery comparison of linac-based SRS treatment techniques currently available for single lesion cranial SRS. In total, two dedicated SRS systems (Novalis Tx, Cyberknife) and a HI-ART TomoTherapy system with six different delivery techniques are evaluated. Four delivery techniques are evaluated on a Novalis Tx system: circular cones, dynamic conformal arcs (DCA), static non-coplanar intensity modulated radiotherapy (NCP-IMRT), and volumetric modulated arc therapy (RapidArc) techniques are compared with intensity modulation based helical Tomotherapy on the HI-ART Tomotherapy system and with non-isocentric, multiple overlapping based robotic radiosurgery using the CyberKnife system. Thirteen patients are retrospectively selected for the study. The target volumes of each patient are transferred to a CT scan of a Lucy phantom (Standard Imaging Inc., Middleton, WI, USA) designed for end-to-end SRS QA. In order to evaluate the plans, several indices scoring the conformality, homogeneity and gradients in the plan are calculated and compared for each of the plans. Finally, to check the clinical deliverability of the plans and the delivery accuracy of different systems, a few targets are delivered on each system. A comparison between planned dose on treatment planning system and dose delivered on Gafchromic EBT film (ISP, Wayne, New Jersey, USA) is carried out by comparing dose beam profiles, isodose lines and by calculating gamma index.

  11. Nanotechnology-Based Drug Delivery Systems for Photodynamic Therapy of Cancer: A Review.

    Science.gov (United States)

    Calixto, Giovana Maria Fioramonti; Bernegossi, Jéssica; de Freitas, Laura Marise; Fontana, Carla Raquel; Chorilli, Marlus

    2016-03-11

    Photodynamic therapy (PDT) is a promising alternative approach for improved cancer treatment. In PDT, a photosensitizer (PS) is administered that can be activated by light of a specific wavelength, which causes selective damage to the tumor and its surrounding vasculature. The success of PDT is limited by the difficulty in administering photosensitizers (PSs) with low water solubility, which compromises the clinical use of several molecules. Incorporation of PSs in nanostructured drug delivery systems, such as polymeric nanoparticles (PNPs), solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs), gold nanoparticles (AuNPs), hydrogels, liposomes, liquid crystals, dendrimers, and cyclodextrin is a potential strategy to overcome this difficulty. Additionally, nanotechnology-based drug delivery systems may improve the transcytosis of a PS across epithelial and endothelial barriers and afford the simultaneous co-delivery of two or more drugs. Based on this, the application of nanotechnology in medicine may offer numerous exciting possibilities in cancer treatment and improve the efficacy of available therapeutics. Therefore, the aim of this paper is to review nanotechnology-based drug delivery systems for photodynamic therapy of cancer.

  12. Nanotechnology-Based Drug Delivery Systems for Photodynamic Therapy of Cancer: A Review

    Directory of Open Access Journals (Sweden)

    Giovana Maria Fioramonti Calixto

    2016-03-01

    Full Text Available Photodynamic therapy (PDT is a promising alternative approach for improved cancer treatment. In PDT, a photosensitizer (PS is administered that can be activated by light of a specific wavelength, which causes selective damage to the tumor and its surrounding vasculature. The success of PDT is limited by the difficulty in administering photosensitizers (PSs with low water solubility, which compromises the clinical use of several molecules. Incorporation of PSs in nanostructured drug delivery systems, such as polymeric nanoparticles (PNPs, solid lipid nanoparticles (SLNs, nanostructured lipid carriers (NLCs, gold nanoparticles (AuNPs, hydrogels, liposomes, liquid crystals, dendrimers, and cyclodextrin is a potential strategy to overcome this difficulty. Additionally, nanotechnology-based drug delivery systems may improve the transcytosis of a PS across epithelial and endothelial barriers and afford the simultaneous co-delivery of two or more drugs. Based on this, the application of nanotechnology in medicine may offer numerous exciting possibilities in cancer treatment and improve the efficacy of available therapeutics. Therefore, the aim of this paper is to review nanotechnology-based drug delivery systems for photodynamic therapy of cancer.

  13. A pharmacokinetic overview of nanotechnology-based drug delivery systems: an ADME-oriented approach.

    Science.gov (United States)

    Hamidi, Mehrdad; Azadi, Amir; Rafiei, Pedram; Ashrafi, Hajar

    2013-01-01

    With the extensive progress in nanotechnology-based drug delivery systems, pharmacokinetic evaluations have gained much attention from researchers as a central part of the study of these systems. Because the fulfillment of any therapeutic goal(s) by a novel drug delivery system requires that the absorption, distribution, metabolism, and excretion (ADME) be considered from the early stages of the system design to the final clinical evaluations, extensive knowledge of the pharmacokinetic aspects related to ADME is a crucial part of research in this field. The main objectives of the nanotechnology-based drug delivery systems from a pharmacokinetic viewpoint are (1) an improved drug-release profile in vivo, (2) enhanced drug absorption, (3) site-directed drug distribution, (4) a modified drug metabolism pattern, (5) prolonged drug residence time in body (e.g., in blood circulation), and (6) delayed and/or decreased renal excretion of the drug. Accordingly, the purpose of the current review is to present an insightful summary of pharmacokinetic analyses of nanotechnology-based drug delivery systems along with a critical review of recent findings.

  14. Silica-based systems for oral delivery of drugs, macromolecules and cells.

    Science.gov (United States)

    Diab, Roudayna; Canilho, Nadia; Pavel, Ileana A; Haffner, Fernanda B; Girardon, Maxime; Pasc, Andreea

    2017-11-01

    According to the US Food and Drug Administration and the European Food Safety Authority, amorphous forms of silica and silicates are generally recognized to be safe as oral delivery ingredients in amounts up to 1500mg per day. Silica is used in the formulation of solid dosage forms, e.g. tablets, as glidant or lubricant. The synthesis of silica-based materials depends on the payload nature, drug, macromolecule or cell, and on the target release (active or passive). In the literature, most of the examples deal with the encapsulation of drugs in mesoporous silica nanoparticles. Still to date limited reports concerning the delivery of encapsulated macromolecules and cells have been reported in the field of oral delivery, despite the multiple promising examples demonstrating the compatibility of the sol-gel route with biological entities, likewise the interest of silica as an oral carrier. Silica diatoms appear as an elegant, cost-effective and promising alternative to synthetic sol-gel-based materials. This review reports the latest advances silica-based systems and discusses the potential benefits and drawbacks of using silica for oral delivery of drugs, macromolecules or cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Carbon Nanotube Arrays for Intracellular Delivery and Biological Applications

    Science.gov (United States)

    Golshadi, Masoud

    Introducing nucleic acids into mammalian cells is a crucial step to elucidate biochemical pathways, modify gene expression in immortalized cells, primary cells, and stem cells, and intoduces new approaches for clinical diagnostics and therapeutics. Current gene transfer technologies, including lipofection, electroporation, and viral delivery, have enabled break-through advances in basic and translational science to enable derivation and programming of embryonic stem cells, advanced gene editing using CRISPR (Clustered regularly interspaced short palindromic repeats), and development of targeted anti-tumor therapy using chimeric antigen receptors in T-cells (CAR-T). Despite these successes, current transfection technologies are time consuming and limited by the inefficient introduction of test molecules into large populations of target cells, and the cytotoxicity of the techniques. Moreover, many cell types cannot be consistently transfected by lipofection or electroporation (stem cells, T-cells) and viral delivery has limitations to the size of experimental DNA that can be packaged. In this dissertation, a novel coverslip-like platform consisting of an array of aligned hollow carbon nanotubes (CNTs) embedded in a sacrificial template is developed that enhances gene transfer capabilities, including high efficiency, low toxicity, in an expanded range of target cells, with the potential to transfer mixed combinations of protein and nucleic acids. The CNT array devices are fabricated by a scalable template-based manufacturing method using commercially available membranes, eliminating the need for nano-assembly. High efficient transfection has been demonstrated by delivering various cargos (nanoparticles, dye and plasmid DNA) into populations of cells, achieving 85% efficiency of plasmid DNA delivery into immortalized cells. Moreover, the CNT-mediated transfection of stem cells shows 3 times higher efficiency compared to current lipofection methods. Evaluating the cell

  16. FFT-based Network Coding For Peer-To-Peer Content Delivery

    OpenAIRE

    Soro, Alexandre; Lacan, Jérôme

    2009-01-01

    In this paper, we propose a structured peer-to-peer (P2P) distribution scheme based on Fast Fourier Transform (FFT) graphs. We build a peer-to-peer network that reproduces the FFT graph initially designed for hardware FFT codecs. This topology allows content delivery with a maximum diversity level for a minimum global complexity. The resulting FFT-based network is a structured architecture with an adapted network coding that brings flexibility upon content distribution and robustness upon the...

  17. Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Nehmé, Reine; Atieh, Carla; Fayad, Syntia; Claude, Bérengère; Chartier, Agnès; Tannoury, Mona; Elleuch, Fatma; Abdelkafi, Slim; Pichon, Chantal; Morin, Philippe

    2017-01-01

    Amino acids play a key role in food analysis, clinical diagnostics, and biochemical research. Capillary electrophoresis with laser-induced fluorescence detection was used for the analysis of several amino acids. Amino acid labeling with fluorescein isothiocyanate was conducted using microwave-assisted derivatization at 80°C (680 W) during only 150 s. Good electrophoretic resolution was obtained using a background electrolyte composed of sodium tetraborate buffer (100 mM; pH 9.4) and β-cyclodextrin (10 mM), and the limits of quantification were 3-30 nM. The developed capillary electrophoresis with laser-induced fluorescence method was used to analyze amino acids in Dunaliella salina green algae grown under different conditions. A simple extraction technique based on electroporation of the cell membrane was introduced. A home-made apparatus allowed the application of direct and alternating voltages across the electrochemical compartment containing a suspension of microalgae in distilled water at 2.5 g/L. A direct voltage of 12 V applied for 4 min gave the optimum extraction yield. Results were comparable to those obtained with accelerated-solvent extraction. The efficiency of electroporation in destroying microalgae membranes was shown by examining the algae surface morphology using scanning electron microscopy. Stress conditions were found to induce the production of amino acids in Dunaliella salina cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Seaweed Polysaccharide-Based Nanoparticles: Preparation and Applications for Drug Delivery

    Directory of Open Access Journals (Sweden)

    Jayachandran Venkatesan

    2016-01-01

    Full Text Available In recent years, there have been major advances and increasing amounts of research on the utilization of natural polymeric materials as drug delivery vehicles due to their biocompatibility and biodegradability. Seaweed polysaccharides are abundant resources and have been extensively studied for several biological, biomedical, and functional food applications. The exploration of seaweed polysaccharides for drug delivery applications is still in its infancy. Alginate, carrageenan, fucoidan, ulvan, and laminarin are polysaccharides commonly isolated from seaweed. These natural polymers can be converted into nanoparticles (NPs by different types of methods, such as ionic gelation, emulsion, and polyelectrolyte complexing. Ionic gelation and polyelectrolyte complexing are commonly employed by adding cationic molecules to these anionic polymers to produce NPs of a desired shape, size, and charge. In the present review, we have discussed the preparation of seaweed polysaccharide-based NPs using different types of methods as well as their usage as carriers for the delivery of various therapeutic molecules (e.g., proteins, peptides, anti-cancer drugs, and antibiotics. Seaweed polysaccharide-based NPs exhibit suitable particle size, high drug encapsulation, and sustained drug release with high biocompatibility, thereby demonstrating their high potential for safe and efficient drug delivery.

  19. Preparation and Characterization of Gelatin-Based Mucoadhesive Nanocomposites as Intravesical Gene Delivery Scaffolds

    Directory of Open Access Journals (Sweden)

    Ching-Wen Liu

    2014-01-01

    Full Text Available This study aimed to develop optimal gelatin-based mucoadhesive nanocomposites as scaffolds for intravesical gene delivery to the urothelium. Hydrogels were prepared by chemically crosslinking gelatin A or B with glutaraldehyde. Physicochemical and delivery properties including hydration ratio, viscosity, size, yield, thermosensitivity, and enzymatic degradation were studied, and scanning electron microscopy (SEM was carried out. The optimal hydrogels (H, composed of 15% gelatin A175, displayed an 81.5% yield rate, 87.1% hydration ratio, 42.9 Pa·s viscosity, and 125.8 nm particle size. The crosslinking density of the hydrogels was determined by performing pronase degradation and ninhydrin assays. In vitro lentivirus (LV release studies involving p24 capsid protein analysis in 293T cells revealed that hydrogels containing lentivirus (H-LV had a higher cumulative release than that observed for LV alone (3.7-, 2.3-, and 2.3-fold at days 1, 3, and 5, resp.. Lentivirus from lentivector constructed green fluorescent protein (GFP was then entrapped in hydrogels (H-LV-GFP. H-LV-GFP showed enhanced gene delivery in AY-27 cells in vitro and to rat urothelium by intravesical instillation in vivo. Cystometrogram showed mucoadhesive H-LV reduced peak micturition and threshold pressure and increased bladder compliance. In this study, we successfully developed first optimal gelatin-based mucoadhesive nanocomposites as intravesical gene delivery scaffolds.

  20. Quantitative Analysis and Modeling of 3-D TSV-Based Power Delivery Architectures

    Science.gov (United States)

    He, Huanyu

    As 3-D technology enters the commercial production stage, it is critical to understand different 3-D power delivery architectures on the stacked ICs and packages with through-silicon vias (TSVs). Appropriate design, modeling, analysis, and optimization approaches of the 3-D power delivery system are of foremost significance and great practical interest to the semiconductor industry in general. Based on fundamental physics of 3-D integration components, the objective of this thesis work is to quantitatively analyze the power delivery for 3D-IC systems, develop appropriate physics-based models and simulation approaches, understand the key issues, and provide potential solutions for design of 3D-IC power delivery architectures. In this work, a hybrid simulation approach is adopted as the major approach along with analytical method to examine 3-D power networks. Combining electromagnetic (EM) tools and circuit simulators, the hybrid approach is able to analyze and model micrometer-scale components as well as centimeter-scale power delivery system with high accuracy and efficiency. The parasitic elements of the components on the power delivery can be precisely modeled by full-wave EM solvers. Stack-up circuit models for the 3-D power delivery networks (PDNs) are constructed through a partition and assembly method. With the efficiency advantage of the SPICE circuit simulation, the overall 3-D system power performance can be analyzed and the 3-D power delivery architectures can be evaluated in a short computing time. The major power delivery issues are the voltage drop (IR drop) and voltage noise. With a baseline of 3-D power delivery architecture, the on-chip PDNs of TSV-based chip stacks are modeled and analyzed for the IR drop and AC noise. The basic design factors are evaluated using the hybrid approach, such as the number of stacked chips, the number of TSVs, and the TSV arrangement. Analytical formulas are also developed to evaluate the IR drop in 3-D chip stack in

  1. Typology of delivery quality: latent profile analysis of teacher engagement and delivery techniques in a school-based prevention intervention, keepin’ it REAL curriculum

    Science.gov (United States)

    Shin, YoungJu; Miller-Day, Michelle; Pettigrew, Jonathan; Hecht, Michael L.; Krieger, Janice L.

    2014-01-01

    Enhancing the delivery quality of school-based, evidence-based prevention programs is one key to ensuring uniform program effects on student outcomes. Program evaluations often focus on content dosage when implementing prevention curricula, however, less is known about implementation quality of prevention content, especially among teachers who may or may not have a prevention background. The goal of the current study is to add to the scholarly literature on implementation quality for a school-based substance use prevention intervention. Twenty-five schools in Ohio and Pennsylvania implemented the original keepin’ REAL (kiR) substance use prevention curriculum. Each of the 10, 40–45 min lessons of the kiR curriculum was video recorded. Coders observed and rated a random sample of 276 videos reflecting 78 classes taught by 31 teachers. Codes included teachers’ delivery techniques (e.g. lecture, discussion, demonstration and role play) and engagement with students (e.g. attentiveness, enthusiasm and positivity). Based on the video ratings, a latent profile analysis was run to identify typology of delivery quality. Five profiles were identified: holistic approach, attentive teacher-orientated approach, enthusiastic lecture approach, engaged interactive learning approach and skill practice-only approach. This study provides a descriptive typology of delivery quality while implementing a school-based substance use prevention intervention. PMID:25274721

  2. Typology of delivery quality: latent profile analysis of teacher engagement and delivery techniques in a school-based prevention intervention, keepin' it REAL curriculum.

    Science.gov (United States)

    Shin, YoungJu; Miller-Day, Michelle; Pettigrew, Jonathan; Hecht, Michael L; Krieger, Janice L

    2014-12-01

    Enhancing the delivery quality of school-based, evidence-based prevention programs is one key to ensuring uniform program effects on student outcomes. Program evaluations often focus on content dosage when implementing prevention curricula, however, less is known about implementation quality of prevention content, especially among teachers who may or may not have a prevention background. The goal of the current study is to add to the scholarly literature on implementation quality for a school-based substance use prevention intervention. Twenty-five schools in Ohio and Pennsylvania implemented the original keepin' REAL (kiR) substance use prevention curriculum. Each of the 10, 40-45 min lessons of the kiR curriculum was video recorded. Coders observed and rated a random sample of 276 videos reflecting 78 classes taught by 31 teachers. Codes included teachers' delivery techniques (e.g., lecture, discussion, demonstration and role play) and engagement with students (e.g. attentiveness, enthusiasm and positivity). Based on the video ratings, a latent profile analysis was run to identify typology of delivery quality. Five profiles were identified: holistic approach, attentive teacher-orientated approach, enthusiastic lecture approach, engaged interactive learning approach and skill practice-only approach. This study provides a descriptive typology of delivery quality while implementing a school-based substance use prevention intervention. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  3. Dual-porosity model of solute diffusion in biological tissue modified by electroporation.

    Science.gov (United States)

    Mahnič-Kalamiza, Samo; Miklavčič, Damijan; Vorobiev, Eugène

    2014-07-01

    In many electroporation applications mass transport in biological tissue is of primary concern. This paper presents a theoretical advancement in the field and gives some examples of model use in electroporation applications. The study focuses on post-treatment solute diffusion. We use a dual-porosity approach to describe solute diffusion in electroporated biological tissue. The cellular membrane presents a hindrance to solute transport into the extracellular space and is modeled as electroporation-dependent porosity, assigned to the intracellular space (the finite rate of mass transfer within an individual cell is not accounted for, for reasons that we elaborate on). The second porosity is that of the extracellular space, through which solute vacates a block of tissue. The model can be used to study extraction out of or introduction of solutes into tissue, and we give three examples of application, a full account of model construction, validation with experiments, and a parametrical analysis. To facilitate easy implementation and experimentation by the reader, the complete derivation of the analytical solution for a simplified example is presented. Validation is done by comparing model results to experimentally-obtained data; we modeled kinetics of sucrose extraction by diffusion from sugar beet tissue in laboratory-scale experiments. The parametrical analysis demonstrates the importance of selected physicochemical and geometrical properties of the system, illustrating possible outcomes of applying the model to different electroporation applications. The proposed model is a new platform that supports rapid extension by state-of-the-art models of electroporation phenomena, developed as latest achievements in the field of electroporation. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis

    Directory of Open Access Journals (Sweden)

    Chambers David

    2011-04-01

    Full Text Available Abstract Background In ovo electroporation is a widely used technique to study gene function in developmental biology. Despite the widespread acceptance of this technique, no genome-wide analysis of the effects of in ovo electroporation, principally the current applied across the tissue and exogenous vector DNA introduced, on endogenous gene expression has been undertaken. Here, the effects of electric current and expression of a GFP-containing construct, via electroporation into the midbrain of Hamburger-Hamilton stage 10 chicken embryos, are analysed by microarray. Results Both current alone and in combination with exogenous DNA expression have a small but reproducible effect on endogenous gene expression, changing the expression of the genes represented on the array by less than 0.1% (current and less than 0.5% (current + DNA, respectively. The subset of genes regulated by electric current and exogenous DNA span a disparate set of cellular functions. However, no genes involved in the regional identity were affected. In sharp contrast to this, electroporation of a known transcription factor, Dmrt5, caused a much greater change in gene expression. Conclusions These findings represent the first systematic genome-wide analysis of the effects of in ovo electroporation on gene expression during embryonic development. The analysis reveals that this process has minimal impact on the genetic basis of cell fate specification. Thus, the study demonstrates the validity of the in ovo electroporation technique to study gene function and expression during development. Furthermore, the data presented here can be used as a resource to refine the set of transcriptional responders in future in ovo electroporation studies of specific gene function.

  5. Factors Associated with Teacher Delivery of a Classroom-Based Tier 2 Prevention Program.

    Science.gov (United States)

    Sutherland, Kevin S; Conroy, Maureen A; McLeod, Bryce D; Algina, James; Kunemund, Rachel L

    2018-02-01

    Teachers sometimes struggle to deliver evidence-based programs designed to prevent and ameliorate chronic problem behaviors of young children with integrity. Identifying factors associated with variations in the quantity and quality of delivery is thus an important goal for the field. This study investigated factors associated with teacher treatment integrity of BEST in CLASS, a tier-2 prevention program designed for young children at risk for developing emotional/behavioral disorders. Ninety-two early childhood teachers and 231 young children at-risk for emotional/behavioral disorders participated in the study. Latent growth curve analyses indicated that both adherence and competence of delivery increased across six observed time points. Results suggest that teacher education and initial levels of classroom quality may be important factors to consider when teachers deliver tier-2 (i.e., targeted to children who are not responsive to universal or tier-1 programming) prevention programs in early childhood settings. Teachers with higher levels of education delivered the program with more adherence and competence initially. Teachers with higher initial scores on the Emotional Support subscale of the Classroom Assessment Scoring System (CLASS) delivered the program with more competence initially and exhibited higher growth in both adherence and competence of delivery across time. Teachers with higher initial scores on the Classroom Organization subscale of the CLASS exhibited lower growth in adherence across time. Contrary to hypotheses, teacher self-efficacy did not predict adherence, and teachers who reported higher initial levels of Student Engagement self-efficacy exhibited lower growth in competence of delivery. Results are discussed in relation to teacher delivery of evidence-based programs in early childhood classrooms.

  6. Spatio-temporal control strategy of drug delivery systems based nano structures.

    Science.gov (United States)

    Rahoui, Nahla; Jiang, Bo; Taloub, Nadia; Huang, Yu Dong

    2017-06-10

    The drug instability, toxicity and the barrier to the target area necessitate a suitable drug delivery system with an external or internal control of the release. Spatio-temporal control using a surface functionalized nano-carrier seems to be the best alternative for guided drug delivery and release. This manuscript provides a broad spectrum about the drug carrier interface modification to cover the need for temporal drug delivery control under neglect side effects. On the other hand, recent advances related to the drug vehicle are highlighted, besides physical (Electric field, magnetic field, light) or mechanical (Ultrasound, mechanical strain), chemical (pH, redox gradient, enzyme) stimuli mediated DDS. Precisely, the paper focus on the NIR light as an effective external stimulus for remotely-triggered DDS. NIR responsive drug delivery systems are considered as novel drug modality that ensures an eco-friendly spatiotemporal control and an administrated meditation. This study also investigated the NIIR spectroscopy (NIRS) combined with partial least square (PLS) for quantitative analysis of a polyvinyl alcohol based prodrug (PVA-DOX) in order to reveal the high potential of NIRS for drug release monitoring and the extraction of concise calibration models. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Recent Advances in Nucleic Acid-Based Delivery: From Bench to Clinical Trials in Genetic Diseases.

    Science.gov (United States)

    Oliveira, Cláudia; Ribeiro, António J; Veiga, Francisco; Silveira, Isabel

    2016-05-01

    Delivery of nucleic acids is the most promising therapy for many diseases that remain untreatable. Therefore, many research efforts have been put on finding a safe and efficient delivery system able to provide a sustained response. Viral vectors have proved to be the most efficient for delivery of nucleic acids and, thus, stand as the foremost vector used in current clinical trials. However, safety issues arise as a main concern and mitigate their use, impelling the improvement of non-viral alternatives. This review focuses on the recent advances in pre-clinical development of non-viral polyplexes and lipoplexes for nucleic acid-based delivery, in contrast with vectors being used in present clinical trials. Nucleic acid vectors for neurodegenerative ataxias, Parkinson's disease, retinitis pigmentosa, cystic fibrosis, hemophilia, pancreatic and lung cancer, and rheumatoid arthritis are discussed to illustrate current state of pre-clinical and clinical studies. Thereby, denoting the prospects for treatment of genetic diseases and elucidating the trend in non-viral vector development and improvement which is expected to significantly increase disease rescue exceeding the modest clinical successes observed so far.

  8. Manufacturing Techniques and Surface Engineering of Polymer Based Nanoparticles for Targeted Drug Delivery to Cancer

    Directory of Open Access Journals (Sweden)

    Yichao Wang

    2016-02-01

    Full Text Available The evolution of polymer based nanoparticles as a drug delivery carrier via pharmaceutical nano/microencapsulation has greatly promoted the development of nano- and micro-medicine in the past few decades. Poly(lactide-co-glycolide (PLGA and chitosan, which are biodegradable and biocompatible polymers, have been approved by both the Food & Drug Administration (FDA and European Medicine Agency (EMA, making them ideal biomaterials that can be advanced from laboratory development to clinical oral and parental administrations. PLGA and chitosan encapsulated nanoparticles (NPs have successfully been developed as new oral drug delivery systems with demonstrated high efficacy. This review aims to provide a comprehensive overview of the fabrication of PLGA and chitosan particulate systems using nano/microencapsulation methods, the current progress and the future outlooks of the nanoparticulate drug delivery systems. Especially, we focus on the formulations and nano/micro-encapsulation techniques using top-down techniques. It also addresses how the different phases including the organic and aqueous ones in the emulsion system interact with each other and subsequently influence the properties of the drug delivery system. Besides, surface modification strategies which can effectively engineer intrinsic physicochemical properties are summarised. Finally, future perspectives and potential directions of PLGA and chitosan nano/microencapsulated drug systems are outlined.

  9. Loop electrosurgical excision of the cervix and subsequent risk for spontaneous preterm delivery: a population-based study of singleton deliveries during a 9-year period

    DEFF Research Database (Denmark)

    Noehr, Bugge; Jensen, Allan; Frederiksen, Kirsten

    2009-01-01

    OBJECTIVE: Our aim was to assess the association between loop electrosurgical excision procedure (LEEP) and the subsequent risk for spontaneous preterm delivery, with the use of population-based data from various nationwide registries. STUDY DESIGN: The study population consisted of all singleton...

  10. Obstetrical nursing care based on good practices: from admission to delivery

    Directory of Open Access Journals (Sweden)

    Maraysa Jéssyca de Oliveira Vieira

    2016-06-01

    Full Text Available The objective of this study is to assess the obstetric nurse’s care from admission to delivery based on good obstetric practices. It is a descriptive, retrospective, and documentary study, with a quantitative approach based on the analysis of 500 medical records. The research identified that an episiotomy was done in 12.2%, an amniotomy in 13%, oxytocin was used in 42.8%, non-pharmacological methods for pain relief were applied in 75.4%, and skin-to-skin contact was used to stimulate in 91.6%. Most of the good obstetrical practices recommended by the World Health Organization were observed to be used by the obstetric nurses when providing care from admission to delivery and this is close to what has been recommended by scientific evidence.

  11. Lipid Nanocapsule-Based Gels for Enhancement of Transdermal Delivery of Ketorolac Tromethamine

    Directory of Open Access Journals (Sweden)

    Jaleh Varshosaz

    2011-01-01

    Full Text Available Previous reports show ineffective transdermal delivery of ketorolac by nanostructured lipid carriers (NLCs. The aim of the present work was enhancement of transdermal delivery of ketorolac by another colloidal carriers, lipid nanocapsules (LNCs. LNCs were prepared by emulsification with phase transition method and mixed in a Carbomer 934P gel base with oleic acid or propylene glycol as penetration enhancers. Permeation studies were performed by Franz diffusion cell using excised rat abdominal skin. Aerosil-induced rat paw edema model was used to investigate the in vivo performance. LNCs containing polyethylene glycol hydroxyl stearate, lecithin in Labrafac as the oily phase, and dilution of the primary emulsion with 3.5-fold volume of cold water produced the optimized nanoparticles. The 1% Carbomer gel base containing 10% oleic acid loaded with nanoparticles enhanced and prolonged the anti-inflammatory effects of this drug to more than 12 h in Aerosil-induced rat paw edema model.

  12. Family Coaching as a delivery modality for evidence-based prevention programs.

    Science.gov (United States)

    Rotheram-Borus, Mary Jane; Swendeman, Dallas; Rotheram-Fuller, Erin; Youssef, Maryann K

    2017-08-01

    Family Coaching is proposed as a new delivery format for evidence-based prevention programs (EBPPs). Three recent developments in health promotion support the potential efficacy of Family Coaching: (1) renewed interest in integrated prevention programs for multiple risk factors and behavior changes, (2) broad and long-term impacts of family-based interventions, and (3) popular acceptance of "coaching" as a nonstigmatizing, goal-focused intervention strategy. Family coaches are community members and paraprofessionals trained in common elements of EBPP. Family Coaching has specific goals, is short term, and has definable outcomes. Coaches frame the program's goals to be consistent with the family's values, normalize the family's experience, assess their strengths, and help the family set goals and develop skills and routines to problem solve challenging situations. Broad dissemination of EBPP will be facilitated with delivery formats that are flexible to meet families' priorities and providers' desires and capacities to tailor programs to local contexts.

  13. Smart Inulin-Based Polycationic Nanodevices for siRNA Delivery.

    Science.gov (United States)

    Cavallaro, G; Sardo, C; Scialabba, C; Licciardi, M; Giammona, G

    2017-01-01

    The advances of short interfering RNA (siRNA) mediated therapy provide a powerful option for the treatment of many diseases by silencing the expression of targeted genes including cancer development and progression. Inulin is a very simple and biocompatible polysaccharide proposed by our groups to produce interesting delivery systems for Nucleic Acid Based Drugs (NABDs), such as siRNA, either as polycations able to give polyplexes and polymeric coatings for nanosystems having a metallic core. In this research field, different functionalizing groups were linked to the inulin backbone with specific aims including oligoamine such as Ethylendiammine (EDA), Diethylediamine (DETA), Spermine, (SPM) etc. In this contribution the main Inulin-based nanodevices for the delivery of siRNA have been reported, analysed and compared with particular reference to their chemical design and structure, biocompatibility, siRNA complexing ability, silencing ability. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Recent Innovations in Peptide Based Targeted Drug Delivery to Cancer Cells

    Directory of Open Access Journals (Sweden)

    Yosi Gilad

    2016-05-01

    Full Text Available Targeted delivery of chemotherapeutics and diagnostic agents conjugated to carrier ligands has made significant progress in recent years, both in regards to the structural design of the conjugates and their biological effectiveness. The goal of targeting specific cell surface receptors through structural compatibility has encouraged the use of peptides as highly specific carriers as short peptides are usually non-antigenic, are structurally simple and synthetically diverse. Recent years have seen many developments in the field of peptide based drug conjugates (PDCs, particularly for cancer therapy, as their use aims to bypass off-target side-effects, reducing the morbidity common to conventional chemotherapy. However, no PDCs have as yet obtained regulatory approval. In this review, we describe the evolution of the peptide-based strategy for targeted delivery of chemotherapeutics and discuss recent innovations in the arena that should lead in the near future to their clinical application.

  15. Cell Based Drug Delivery: Micrococcus luteus Loaded Neutrophils as Chlorhexidine Delivery Vehicles in a Mouse Model of Liver Abscesses in Cattle.

    Directory of Open Access Journals (Sweden)

    Sebastian O Wendel

    Full Text Available The recent WHO report on antibiotic resistances shows a dramatic increase of microbial resistance against antibiotics. With only a few new antibiotics in the pipeline, a different drug delivery approach is urgently needed. We have obtained evidence demonstrating the effectiveness of a cell based drug delivery system that utilizes the innate immune system as targeting carrier for antibacterial drugs. In this study we show the efficient loading of neutrophil granulocytes with chlorhexidine and the complete killing of E. coli as well as Fusobacterium necrophorum in in-vitro studies. Fusobacterium necrophorum causes hepatic abscesses in cattle fed high grain diets. We also show in a mouse model that this delivery system targets infections of F. necrophorum in the liver and reduces the bacterial burden by an order of magnitude from approximately 2•106 to 1•105.

  16. Cell Based Drug Delivery: Micrococcus luteus Loaded Neutrophils as Chlorhexidine Delivery Vehicles in a Mouse Model of Liver Abscesses in Cattle.

    Science.gov (United States)

    Wendel, Sebastian O; Menon, Sailesh; Alshetaiwi, Hamad; Shrestha, Tej B; Chlebanowski, Lauren; Hsu, Wei-Wen; Bossmann, Stefan H; Narayanan, Sanjeev; Troyer, Deryl L

    2015-01-01

    The recent WHO report on antibiotic resistances shows a dramatic increase of microbial resistance against antibiotics. With only a few new antibiotics in the pipeline, a different drug delivery approach is urgently needed. We have obtained evidence demonstrating the effectiveness of a cell based drug delivery system that utilizes the innate immune system as targeting carrier for antibacterial drugs. In this study we show the efficient loading of neutrophil granulocytes with chlorhexidine and the complete killing of E. coli as well as Fusobacterium necrophorum in in-vitro studies. Fusobacterium necrophorum causes hepatic abscesses in cattle fed high grain diets. We also show in a mouse model that this delivery system targets infections of F. necrophorum in the liver and reduces the bacterial burden by an order of magnitude from approximately 2•106 to 1•105.

  17. A Transdermal Drug Delivery System Based on LIGA Technology and Soft Lithography

    Science.gov (United States)

    Matteucci, Marco; Perennes, Frederic; Marmiroli, Benedetta; Di Fabrizio, Enzo

    2007-01-01

    This report presents a transdermal drug delivery system based on LIGA fabricated microparts. It is a portable device combining a magnetically actuated micro gear pump with a microneedle array. The fluidic behaviour of the system is analyzed in order to predict its performance according to the dimension of the microparts and then compared to experimental data. The manufacturing process of both micropump and microneedle array are described.

  18. Recent Trends in Nanotechnology-Based Drugs and Formulations for Targeted Therapeutic Delivery.

    Science.gov (United States)

    Iqbal, Hafiz M N; Rodriguez, Angel M V; Khandia, Rekha; Munjal, Ashok; Dhama, Kuldeep

    2017-01-01

    In the recent past, a wider spectrum of nanotechnologybased drugs or drug-loaded devices and systems has been engineered and investigated with high interests. The key objective is to help for an enhanced/better quality of patient life in a secure way by avoiding/limiting drug abuse, or severe adverse effects of some in practice traditional therapies. Various methodological approaches including in vitro, in vivo, and ex vivo techniques have been exploited, so far. Among them, nanoparticles-based therapeutic agents are of supreme interests for an enhanced and efficient delivery in the current biomedical sector of the modern world. The development of new types of novel, effective and highly reliable therapeutic drug delivery system (DDS) for multipurpose applications is essential and a core demand to tackle many human health related diseases. In this context, nanotechnology-based several advanced DDS have been engineered with novel characteristics for biomedical, pharmaceutical and cosmeceutical applications that include but not limited to the enhanced/improved bioactivity, bioavailability, drug efficacy, targeted delivery, and therapeutically safer with an extra advantage of overcoming demerits of traditional drug formulations/designs. This review work is focused on recent trends/advances in nanotechnology-based drugs and formulations designed for targeted therapeutic delivery. Moreover, information is also reviewed and given from recent patents and summarized or illustrated diagrammatically to depict a better understanding. Recent patents covering various nanotechnology-based approaches for several applications have also been reviewed. The drug-loaded nanoparticles are among versatile candidates with multifunctional characteristics for potential applications in biomedical, and tissue engineering sector. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  19. The intracellular trafficking mechanism of Lipofectamine-based transfection reagents and its implication for gene delivery

    OpenAIRE

    Francesco Cardarelli; Luca Digiacomo; Cristina Marchini; Augusto Amici; Fabrizio Salomone; Giuseppe Fiume; Alessandro Rossetta; Enrico Gratton; Daniela Pozzi; Giulio Caracciolo

    2016-01-01

    Lipofectamine reagents are widely accepted as ?gold-standard? for the safe delivery of exogenous DNA or RNA into cells. Despite this, a satisfactory mechanism-based explanation of their superior efficacy has remained mostly elusive thus far. Here we apply a straightforward combination of live cell imaging, single-particle tracking microscopy, and quantitative transfection-efficiency assays on live cells to unveil the intracellular trafficking mechanism of Lipofectamine/DNA complexes. We find ...

  20. pH-responsive alginate-based composite hydrogel beads for oral insulin delivery

    OpenAIRE

    HUI PENG LIM

    2017-01-01

    This research attempts to develop a pH-responsive alginate-based composite hydrogel beads for encapsulation and controlled delivery of oral insulin aspart through incorporation of κ-carrageenan as a reinforcement polymer and hydrophobic modification of alginate polymer. The alginate/κ-carrageenan composite hydrogel beads were able to fully retain the insulin in conditions mimicking the stomach and release the insulin in conditions mimicking the intestine at a constant rate. Bioactivity of ins...

  1. 29B. Enhancing Primary Healthcare Delivery in the Inner City Through Interprofessional Team-based Care

    OpenAIRE

    Kopansky-Giles, Deborah; Leung, Fok-Han

    2013-01-01

    Focus Area: Sustainable Business Models This session will describe the evolution of an integrative model of primary healthcare delivery within a hospital-based setting. Over the past decade, the Academic Family Health Team (AFHT) in the Department of Family and Community Medicine at St Michael's Hospital in Toronto has been in the business of transforming primary healthcare. The AFHT provides services to the inner-city community of Toronto through 5 clinical sites located conveniently in the ...

  2. Designing a Mobile Based Library Information and Service Delivery System: A Model Plan for Libraries of Bangladesh

    Directory of Open Access Journals (Sweden)

    Elahi, Md. Hasinul

    2016-09-01

    Full Text Available Purpose The purpose of the study is to identify the need of a mobile based library information and service delivery system in developing countries, identify the services that are possible to provide with the help of mobile phones and their applications, design a mobile based library information and service delivery system, and identify major challenges regarding mobile based library information and service delivery systems.Design/ Methodology/ Approach The research is exploratory in nature. This paper is based on a review of literature, survey information, and on the author’s own viewpoints. A comprehensive search of scientific literature has been done along with a survey for validating the necessity of designing mobile based library information and service delivery systems. Responses to closed-ended questions in particular on 7-point Likert scales were analyzed using the descriptive analysis techniques of SPSS 20.0 and responses to other closed-ended questions were analyzed using general statistics.Findings The research indicates that users of academic libraries, especially students and faculty members, have a positive notion regarding designing and implementing a mobile based library information and service delivery system. It also reveals user opinions regarding the possible problems associated with the design and implementation of a mobile based library information and service delivery system.Originality The paper explores mobile based library information and service delivery systems for better and faster service provision to its potential users.

  3. Irreversible electroporation of human primary uveal melanoma in enucleated eyes.

    Directory of Open Access Journals (Sweden)

    Yossi Mandel

    Full Text Available Uveal melanoma (UM is the most common primary intraocular tumor in adults and is characterized by high rates of metastatic disease. Although brachytherapy is the most common globe-sparing treatment option for small- and medium-sized tumors, the treatment is associated with severe adverse reactions and does not lead to increased survival rates as compared to enucleation. The use of irreversible electroporation (IRE for tumor ablation has potential advantages in the treatment of tumors in complex organs such as the eye. Following previous theoretical work, herein we evaluate the use of IRE for uveal tumor ablation in human ex vivo eye model. Enucleated eyes of patients with uveal melanoma were treated with short electric pulses (50-100 µs, 1000-2000 V/cm using a customized electrode design. Tumor bioimpedance was measured before and after treatment and was followed by histopathological evaluation. We found that IRE caused tumor ablation characterized by cell membrane disruption while sparing the non-cellular sclera. Membrane disruption and loss of cellular capacitance were also associated with significant reduction in total tumor impedance and loss of impedance frequency dependence. The effect was more pronounced near the pulsing electrodes and was dependent on time from treatment to fixation. Future studies should further evaluate the potential of IRE as an alternative method of uveal melanoma treatment.

  4. Irreversible electroporation in primary and metastatic hepatic malignancies: A review.

    Science.gov (United States)

    Lyu, Tianchu; Wang, Xifu; Su, Zhanliang; Shangguan, Junjie; Sun, Chong; Figini, Matteo; Wang, Jian; Yaghmai, Vahid; Larson, Andrew C; Zhang, Zhuoli

    2017-04-01

    Liver cancer makes up a huge percentage of cancer mortality worldwide. Irreversible electroporation (IRE) is a relatively new minimally invasive nonthermal ablation technique for tumors that applies short pulses of high frequency electrical energy to irreversibly destabilize cell membrane to induce tumor cell apoptosis. This review aims to investigate the studies regarding the use of IRE treatment in liver tumors and metastases to liver. We searched PubMed for all of IRE relevant English language articles published up to September 2016. They included clinical trials, experimental studies, observational studies, and reviews. This review manuscript is nothing with ethics issues and ethical approval is not provided. In recent years, increasingly more studies in both preclinical and clinical settings have been conducted to examine the safety and efficacy of this new technique, shedding light on the crucial advantages and disadvantages that IRE possesses. Unlike the current leading thermal ablation techniques, such as radiofrequency ablation (RFA), microwave ablation (MWA), and cryoablation, IRE requires shorter ablation time without damaging adjacent important vital structures. Although IRE has successfully claimed its valuable status in the field of hepatic cancer treatment both preclinical and clinical settings. In order to systemically test and establish its safety and efficacy for clinical applications, more studies still need to be conducted.

  5. The effects of irreversible electroporation (IRE on nerves.

    Directory of Open Access Journals (Sweden)

    Wei Li

    Full Text Available BACKGROUND: If a critical nerve is circumferentially involved with tumor, radical surgery intended to cure the cancer must sacrifice the nerve. Loss of critical nerves may lead to serious consequences. In spite of the impressive technical advancements in nerve reconstruction, complete recovery and normalization of nerve function is difficult to achieve. Though irreversible electroporation (IRE might be a promising choice to treat tumors near or involved critical nerve, the pathophysiology of the nerve after IRE treatment has not be clearly defined. METHODS: We applied IRE directly to a rat sciatic nerve to study the long term effects of IRE on the nerve. A sequence of 10 square pulses of 3800 V/cm, each 100 µs long was applied directly to rat sciatic nerves. In each animal of group I (IRE the procedure was applied to produce a treated length of about 10 mm. In each animal of group II (Control the electrodes were only applied directly on the sciatic nerve for the same time. Electrophysiological, histological, and functional studies were performed on immediately after and 3 days, 1 week, 3, 5, 7 and 10 weeks following surgery. FINDINGS: Electrophysiological, histological, and functional results show the nerve treated with IRE can attain full recovery after 7 weeks. CONCLUSION: This finding is indicative of the preservation of nerve involving malignant tumors with respect to the application of IRE pulses to ablate tumors completely. In summary, IRE may be a promising treatment tool for any tumor involving nerves.

  6. A population based evaluation of the mode of delivery in association with infertility treatment from 1990-2012.

    Science.gov (United States)

    Reichelt, J; Kyvernitakis, I; Misselwitz, B; Hadji, P; Schmidt, S; Kalder, M

    2015-02-01

    This study refers to population based data and investigates the development of the mode of delivery associated with infertility treatment over the last 23 years. All 1 202,557 deliveries in Hesse, Germany, between 1990 and 2012 were assessed. 2.2% of the study population, 26,761, had a delivery subsequent to infertility treatment based on the Hessian Perinatal Registry (HEPE). An evaluation in this subgroup was performed investigating the associations between the mode of delivery and the gestational week and the mother's age. A continuous and significant (pinfertility treatment (1990: 41,3%; 2012: 55,9%) as well as a conversely also significant (pinfertility treatment between 1990 and 2012 was found. Furthermore, the preterm delivery rate and the proportion of deliveries of parturients older than 35 years of age in association with infertility treatment raised over the last years. Rates of full-term deliveries and deliveries of women younger than 35 years remained stable during the observation period. The rate of cesarean section is continuously rising over the last 23 years with regard to parturients subsequent to infertility treatment. The CS rate is significantly higher compared to women with a spontaneous pregnancy and in comparison to the data from 20 years ago. Most recently, the number of CS (51,2%) exceeded the number of vaginal deliveries (48,8%) in Hesse subsequent to infertility treatment for the first time. © Georg Thieme Verlag KG Stuttgart · New York.

  7. Irreversible Electroporation in the Liver: Contrast-enhanced Inversion-Recovery MR Imaging Approaches to Differentiate Reversibly Electroporated Penumbra from Irreversibly Electroporated Ablation Zones

    Science.gov (United States)

    Guo, Yang; Zhang, Yue; Nijm, Grace M.; Sahakian, Alan V.; Yang, Guang-Yu; Omary, Reed A.

    2011-01-01

    Purpose: To evaluate the use of contrast material–enhanced magnetic resonance (MR) imaging with conventional T1-weighted gradient-recalled echo (GRE) and inversion-recovery (IR)-prepared GRE methods to quantitatively measure the size of irreversible electroporation (IRE) ablation zones in the liver in a rat model. Materials and Methods: All studies were approved by the institutional animal care and use committee and were performed in accordance with institutional guidelines. Seventeen adult male Sprague-Dawley rats were divided into four groups. Rats in groups 1–3 (n = 15 total) underwent IRE performed by using different IRE parameters after gadopentetate dimeglumine administration. Rats in group 4 (n = 2) underwent IRE ablation without prior gadopentetate dimeglumine injection to serve as control animals. MR imaging measurements (with conventional T1-weighted GRE and IR-prepared GRE methods) were performed 2 hours after IRE to predict the IRE ablation zones, which were correlated with pathology-confirmed necrosis areas 24 hours after IRE by using the Spearman correlation coefficient. Bland-Altman plots were also generated to investigate the agreement between MR imaging–measured ablation zones and reference standard histologic measurements of corresponding ablation zones. Results: The necrotic areas measured on the pathology images were well correlated with the hyperintense regions measured on T1-weighted GRE images (r = 0.891, P < .001) and normal tissue–nulled IR images (r = 0.874, P < .001); pathology measurements were also well correlated with the smaller hyperintense regions measured on those IR images with inversion times specifically selected to null signal from the peripheral penumbra surrounding the ablation zone (r = 0.939, P < .001). Bland-Altman plots indicated that these penumbra-nulled IR images provided more accurate predictions of IRE ablation zones, with T1-weighted GRE measurements tending to overestimate ablation zone sizes. Conclusion

  8. Exploring information systems outsourcing in U.S. hospital-based health care delivery systems.

    Science.gov (United States)

    Diana, Mark L

    2009-12-01

    The purpose of this study is to explore the factors associated with outsourcing of information systems (IS) in hospital-based health care delivery systems, and to determine if there is a difference in IS outsourcing activity based on the strategic value of the outsourced functions. IS sourcing behavior is conceptualized as a case of vertical integration. A synthesis of strategic management theory (SMT) and transaction cost economics (TCE) serves as the theoretical framework. The sample consists of 1,365 hospital-based health care delivery systems that own 3,452 hospitals operating in 2004. The findings indicate that neither TCE nor SMT predicted outsourcing better than the other did. The findings also suggest that health care delivery system managers may not be considering significant factors when making sourcing decisions, including the relative strategic value of the functions they are outsourcing. It is consistent with previous literature to suggest that the high cost of IS may be the main factor driving the outsourcing decision.

  9. Topical cream-based dosage forms of the macrocyclic drug delivery vehicle cucurbit[6]uril.

    Directory of Open Access Journals (Sweden)

    Marian Seif

    Full Text Available The macrocycle family of molecules called cucurbit[n]urils are potential drug delivery vehicles as they are able to form host-guest complexes with many different classes of drugs. This study aimed to examine the utility of Cucurbit[6]uril (CB[6] in topical cream-based formulations for either localised treatment or for transdermal delivery. Cucurbit[6]uril was formulated into both buffered cream aqueous- and oily cream-based dosage forms. The solid state interaction of CB[6] with other excipients was studied by differential scanning calorimetry and the macrocycle's transdermal permeability was determined using rat skin. Significant solid state interactions were observed between CB[6] and the other dosage form excipients. At concentrations up to 32% w/w the buffered aqueous cream maintained its normal consistency and could be effectively applied to skin, but the oily cream was too stiff and is not suitable as a dosage form. Cucurbit[6]uril does not permeate through skin; as such, the results imply that cucurbituril-based topical creams may potentially only have applications for localised skin treatment and not for transdermal drug delivery.

  10. Topical Cream-Based Dosage Forms of the Macrocyclic Drug Delivery Vehicle Cucurbit[6]uril

    Science.gov (United States)

    Seif, Marian; Impelido, Michael L.; Apps, Michael G.; Wheate, Nial J.

    2014-01-01

    The macrocycle family of molecules called cucurbit[n]urils are potential drug delivery vehicles as they are able to form host-guest complexes with many different classes of drugs. This study aimed to examine the utility of Cucurbit[6]uril (CB[6]) in topical cream-based formulations for either localised treatment or for transdermal delivery. Cucurbit[6]uril was formulated into both buffered cream aqueous- and oily cream-based dosage forms. The solid state interaction of CB[6] with other excipients was studied by differential scanning calorimetry and the macrocycle's transdermal permeability was determined using rat skin. Significant solid state interactions were observed between CB[6] and the other dosage form excipients. At concentrations up to 32% w/w the buffered aqueous cream maintained its normal consistency and could be effectively applied to skin, but the oily cream was too stiff and is not suitable as a dosage form. Cucurbit[6]uril does not permeate through skin; as such, the results imply that cucurbituril-based topical creams may potentially only have applications for localised skin treatment and not for transdermal drug delivery. PMID:24454850

  11. Telemetric light delivery and monitoring system for photodynamic therapy based on solid-state optodes

    Science.gov (United States)

    Margallo-Balbás, Eduardo; Kaptein, Johan G.; Sterenborg, Henricus J. C. M.; Pandraud, Grégory; French, Patrick J.; Robinson, Dominic J.

    2008-02-01

    Light delivery and optical monitoring during photodynamic therapy (PDT) is often limited by the need for a physical optical link between the light source and detection devices and the treatment volume. This can be critical when sources need to be implanted within the body for extended periods. We report on the latest developments for a telemetric PDT delivery and monitoring device that can dynamically vary the local illumination parameters based on the in-situ fluence rate within the PDT target volume. Local light delivery and collection is achieved using solid-state optodes, microfabricated on a silicon substrate. Photodiodes have been produced using a standard bipolar process. Chip-form LEDs are then assembled into micro-machined pits adjacent to the light fluence rate detectors. The devices (1.2×1.2mm2) are bonded to a flexible PCB together with the remaining electronics. Power coupling and communications are achieved by means of an inductive link while light delivery and fluence rate monitoring are digitally managed using a microcontroller. These devices are being tested in optical phantoms and in pre-clinical models. Our results show that it is possible to manufacture solid-state optodes of suitable dimensions and that it is feasible to telemetrically deliver and control the local fluence rate using them. It can also be concluded from our work that while the optode is sufficiently small to be useful as a light delivery and monitoring device, digital control, read-out electronics and power coupling can benefit from further optimization and miniaturization.

  12. Utilization of health care services by pregnant mothers during delivery: a community based study in Nigeria.

    Science.gov (United States)

    Moore, B M; Alex-Hart, B A; George, I O

    2011-03-01

    Poor utilization of health facilities during delivery by pregnant mothers is still a major cause of maternal and childhood morbidity and mortality in Nigeria. The aim of this study was to determine the level of utilization of health care services by pregnant women during delivery in Gokana Local Government Area of River State, Nigeria. This was a cross-sectional, questionnaire; based study involving 112 mothers aged 15 years to 49 years from Gokana Local Government Area of Rivers State, Nigeria. The local Government Area has 12 health centres and 6 health centres were selected by multistage sampling. 112 were then selected by simple random sampling. Of the 112 mothers interviewed 91 (81.3%) were married, 13 (11.6%) were single, 5 (4.5%) were widows, 2 (1.8%) divorced and 1 (0.9%) separated. Ninety seven (86.6%) of these mothers (n = 112) had formal education while 15 (13.4%) had no formal education. Most 42 (37.5%) of the mothers were between 25-29 years. Sixty four (57.1%) of the 112 mothers in their recent delivery used a health facility while 48 (42.9%) did not. Factors responsible for non utilization of health facility for delivery include: Long distance to health facility 33 (68.7%), onset of labour at night 40 (83.3%), unavailability of means of transportation 37 (77.1%), Lack of money for transportation 26 (54.2%), unsatisfactory services at health facility 26 (54.2%), unfriendly attitude of staff of the health facility 34 (70.8%), unavailability of staff at health facility 32 (64.0%), lack of urgency at health facility 36 (75.0%), previous uneventful delivery at the health facility 32 (66.7%). Utilization of health care services during delivery in Nigeria is still poor. Concerted efforts should be made both at community and Government levels to improve utilization of health facility during delivery. This will go a long way in reducing maternal and child mortality.

  13. Optimization of single-cell electroporation protocol for forced gene expression in primary neuronal cultures.

    Science.gov (United States)

    Nishikawa, Shin; Hirashima, Naohide; Tanaka, Masahiko

    2014-09-01

    The development and function of the central nervous system (CNS) are realized through interactions between many neurons. To investigate cellular and molecular mechanisms of the development and function of the CNS, it is thus crucial to be able to manipulate the gene expression of single neurons in a complex cell population. We recently developed a technique for gene silencing by introducing small interfering RNA into single neurons in primary CNS cultures using single-cell electroporation. However, we had not succeeded in forced gene expression by introducing expression plasmids using single-cell electroporation. In the present study, we optimized the experimental conditions to enable the forced expression of green fluorescent protein (GFP) in cultured cerebellar Purkinje neurons using single-cell electroporation. We succeeded in strong GFP expression in Purkinje neurons by increasing the inside diameter of micropipettes or by making the size of the original plasmid smaller by digestion and cyclizing it by ligation. Strong GFP expression in Purkinje neurons electroporated under the optimal conditions continued to be observed for more than 25 days after electroporation. Thus, this technique could be used for forced gene expression in single neurons to investigate cellular and molecular mechanisms of the development, function, and disease of the CNS.

  14. New mechanisms for non-porative ultrasound stimulation of cargo delivery to cell cytosol with targeted perfluorocarbon nanoparticles

    OpenAIRE

    Soman, NR; Marsh, JN; Lanza, GM; Wickline, SA

    2008-01-01

    The cell membrane constitutes a major barrier for non-endocytotic intracellular delivery of therapeutic molecules from drug delivery vehicles. Existing approaches to breaching the cell membrane include cavitational ultrasound (with microbubbles), electroporation and cell-penetrating peptides. We report the use of diagnostic ultrasound for intracellular delivery of therapeutic bulky cargo with the use of molecularly targeted liquid perfluorocarbon (PFC) nanoparticles. To demonstrate the concep...

  15. A new technique for reversible permeabilization of live cells for intracellular delivery of quantum dots

    Science.gov (United States)

    Medepalli, Krishnakiran; Alphenaar, Bruce W.; Keynton, Robert S.; Sethu, Palaniappan

    2013-05-01

    A major challenge with the use of quantum dots (QDs) for cellular imaging and biomolecular delivery is the attainment of QDs freely dispersed inside the cells. Conventional methods such as endocytosis, lipids based delivery and electroporation are associated with delivery of QDs in vesicles and/or as aggregates that are not monodispersed. In this study, we demonstrate a new technique for reversible permeabilization of cells to enable the introduction of freely dispersed QDs within the cytoplasm. Our approach combines osmosis driven fluid transport into cells achieved by creating a hypotonic environment and reversible permeabilization using low concentrations of cell permeabilization agents like Saponin. Our results confirm that highly efficient endocytosis-free intracellular delivery of QDs can be accomplished using this method. The best results were obtained when the cells were treated with 50 μg ml-1 Saponin in a hypotonic buffer at a 3:2 physiological buffer:DI water ratio for 5 min at 4 ° C.

  16. Delivery devices for exposure of biological cells to nanosecond pulsed electric fields.

    Science.gov (United States)

    Soueid, Malak; Dobbelaar, Martinus C F; Bentouati, Sabrina; Bardet, Sylvia M; O'Connor, Rodney P; Bessières, Delphine; Paillol, Jean; Leveque, Philippe; Arnaud-Cormos, Delia

    2018-01-01

    In this paper, delivery devices for nanosecond pulsed electric field exposure of biological samples in direct contact with electrodes or isolated are presented and characterized. They are based on a modified electroporation cuvette and two transverse electromagnetic cells (TEM cells). The devices were used to apply pulses with high intensity (4.5 kV) and short durations (3 and 13 ns). The delivery devices were electromagnetically characterized in the frequency and time domains. Field intensities of around 5, 0.5, and 12 MV m-1 were obtained by numerical simulations of the biological sample positioned in the three delivery devices. Two delivery systems had a homogenous electric field spatial distribution, and one was adapted to permit a highly localized exposure in the vicinity of a needle. Experimental biological investigations were carried out at different field intensities for five cancer cell lines. The results using flow cytometry showed that cells kept polarized mitochondrial membrane but lost plasma membrane integrity following a dose-response trend after exposure to different electric field intensities. Certain cell types (U87, MCF7) showed higher sensitivities to nsPEFs than other lines tested.

  17. Noninvasive measurement of transdermal drug delivery by impedance spectroscopy

    OpenAIRE

    Arpaia, Pasquale; Cesaro, Umberto; Moccaldi, Nicola

    2017-01-01

    The effectiveness in transdermal delivery of skin permeation strategies (e.g., chemical enhancers, vesicular carrier systems, sonophoresis, iontophoresis, and electroporation) is poorly investigated outside of laboratory. In therapeutic application, the lack of recognized techniques for measuring the actually-released drug affects the scientific concept itself of dosage for topically- and transdermally-delivered drugs. Here we prove the suitability of impedance measurement for assessing the a...

  18. Perceptions Toward Internet-Based Delivery of Hearing Aids among Older Hearing-Impaired Adults.

    Science.gov (United States)

    Chandra, Navshika; Searchfield, Grant D

    2016-06-01

    Despite evidence that hearing aids can improve the social and psychological functioning of older hearing-impaired adults, hearing aid uptake is low. High cost of hearing aids and poor access to audiology services in rural areas are potential barriers to hearing aid acquisition. Methods of hearing aid delivery deviating from the traditional clinician-based model have been available to consumers for many years. One such method is Internet hearing aid sales. However, research exploring Internet-based hearing aid delivery, as a method to improve hearing aid uptake in this population, is limited. The purpose of this study was to explore the perceptions of older hearing aid users (aged ≥65 yr) toward Internet-based hearing aid delivery. A qualitative approach was adopted to investigate older adults' perceptions of buying hearing aids online. The sample consisted of 18 participants aged between 64 and 81 yr. Fourteen men and four women participated in this study. Participants were all experienced hearing aid users. Face-to-face semistructured interviews were conducted. An interview schedule guided the interview. Interviews were recorded with a voice recorder and transcribed verbatim. Thematic analysis of the data was carried out. Seven main themes emerged from the data. A general lack of awareness, but willingness to learn more about Internet hearing aid sales, was found. Two perceived benefits of Internet-based hearing aid delivery were identified: lower cost of hearing aids and greater convenience or physical accessibility. Numerous concerns and limitations were communicated. Concerns regarding the availability of clinical procedures, such as hearing tests, obtaining the correct-sized earmolds, and fine-tuning of hearing aids, were expressed. Participants conveyed distrust in online retailers. However, trust in and a preference for audiologists' expertise, which was not perceived to be available online, was found. Participants further conveyed a preference for face

  19. Magnetic Nanoparticle Based Nonviral MicroRNA Delivery into Freshly Isolated CD105+ hMSCs

    Directory of Open Access Journals (Sweden)

    Anna Schade

    2014-01-01

    Full Text Available Genetic modifications of bone marrow derived human mesenchymal stem cells (hMSCs using microRNAs (miRs may be used to improve their therapeutic potential and enable innovative strategies in tissue regeneration. However, most of the studies use cultured hMSCs, although these can lose their stem cell characteristics during expansion. Therefore, we aimed to develop a nonviral miR carrier based on polyethylenimine (PEI bound to magnetic nanoparticles (MNPs for efficient miR delivery in freshly isolated hMSCs. MNP based transfection is preferable for genetic modifications in vivo due to improved selectivity, safety of delivery, and reduced side effects. Thus, in this study different miR/PEI and miR/PEI/MNP complex formulations were tested in vitro for uptake efficiency and cytotoxicity with respect to the influence of an external magnetic field. Afterwards, optimized magnetic complexes were selected and compared to commercially available magnetic vectors (Magnetofectamine, CombiMag. We found that all tested transfection reagents had high miR uptake rates (yielded over 60% and no significant cytotoxic effects. Our work may become crucial for virus-free introduction of therapeutic miRs as well as other nucleic acids in vivo. Moreover, in the field of targeted stem cell therapy nucleic acid delivery prior to transplantation may allowfor initial cell modulation in vitro.

  20. Micro- and nano bio-based delivery systems for food applications: In vitro behavior.

    Science.gov (United States)

    de Souza Simões, Lívia; Madalena, Daniel A; Pinheiro, Ana C; Teixeira, José A; Vicente, António A; Ramos, Óscar L

    2017-05-01

    Micro- and nanoencapsulation is an emerging technology in the food field that potentially allows the improvement of food quality and human health. Bio-based delivery systems of bioactive compounds have a wide variety of morphologies that influence their stability and functional performance. The incorporation of bioactive compounds in food products using micro- and nano-delivery systems may offer extra health benefits, beyond basic nutrition, once their encapsulation may provide protection against undesired environmental conditions (e.g., heat, light and oxygen) along the food chain (including processing and storage), thus improving their bioavailability, while enabling their controlled release and target delivery. This review provides an overview of the bio-based materials currently used for encapsulation of bioactive compounds intended for food applications, as well as the main production techniques employed in the development of micro- and nanosystems. The behavior of such systems and of bioactive compounds entrapped into, throughout in vitro gastrointestinal systems, is also tracked in a critical manner. Comparisons between various in vitro digestion systems (including the main advantages and disadvantages) currently in use, as well as correlations between the behavior of micro- and nanosystems studied through in vitro and in vivo systems were highlighted and discussed here for the first time. Finally, examples of bioactive micro- and nanosystems added to food simulants or to real food matrices are provided, together with a revision of the main challenges for their safe commercialization, the regulatory issues involved and the main legislation aspects. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Microneedle arrays delivery of the conventional vaccines based on nonvirulent viruses.

    Science.gov (United States)

    Li, Ning; Wang, Ning; Wang, Xueting; Zhen, Yuanyuan; Wang, Ting

    2016-11-01

    Recently, microneedle arrays (MAs) have been developed for painless inoculation of vaccines and possess many prominent advantages, including convenience for inoculation, and exact delivery of vaccine to the exact epidermal and dermal or mucosal compartments which teem with antigen-presenting cells (APCs). Among different types of MAs, while the micro-environmental stimulus-responsive MAs represent one of the developmental trends in the field, the MAs combined with the conventional vaccines that are based on nonvirulent viruses, such as live attenuated or whole inactivated viruses, and antigen-encoding DNA viral vectors, have developed rapidly into the advanced stages, with certain products already on clinical trials. The pre- and clinical research outcomes showed that the painless MA delivery of the conventional vaccines through mammalian skin or mucosa can not only elicit robust systemic and even mucosal immunity to pathogens but also, in certain circumstances, redirect the immune response toward a specific Th1 pathway, resulting in cytotoxic T lymphocytes (CTL) to erase the cell-hidden pathogens, thanks to the robust adjuvant function of MAs exerted through damaging the contacted cells to release dangerous signals. This paper focuses on reviewing the latest research and advancements in MA delivery of the conventional vaccines that are based on nonvirulent viruses, underlining MA enhancement of the overall vaccine performance and the most advanced MA vaccine products that are relatively close to markets.

  2. Supramolecular Drug Delivery Systems Based on Water-Soluble Pillar[n]arenes.

    Science.gov (United States)

    Wu, Xuan; Gao, Lei; Hu, Xiao-Yu; Wang, Leyong

    2016-06-01

    Supramolecular drug delivery systems (SDDSs), including various kinds of nanostructures that are assembled by reversible noncovalent interactions, have attracted considerable attention as ideal drug carriers owing to their fascinating ability to undergo dynamic switching of structure, morphology, and function in response to various external stimuli, which provides a flexible and robust platform for designing and developing functional and smart supramolecular nano-drug carriers. Pillar[n]arenes represent a new generation of macrocyclic hosts, which have unique structures and excellent properties in host-guest chemistry. This account describes recent progress in our group to develop pillararene-based stimuli-responsive supramolecular nanostructures constructed by reversible host-guest interactions for controllable anticancer drug delivery. The potential applications of these supramolecular drug carriers in cancer treatment and the fundamental questions facing SDDSs are also discussed. © 2016 The Chemical Society of Japan & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Nanotechnology-based drug delivery systems for treatment of oral cancer: a review.

    Science.gov (United States)

    Calixto, Giovana; Bernegossi, Jéssica; Fonseca-Santos, Bruno; Chorilli, Marlus

    2014-01-01

    Oral cancer (oral cavity and oropharynx) is a common and aggressive cancer that invades local tissue, can cause metastasis, and has a high mortality rate. Conventional treatment strategies, such as surgery and chemoradiotherapy, have improved over the past few decades; however, they remain far from optimal. Currently, cancer research is focused on improving cancer diagnosis and treatment methods (oral cavity and oropharynx) nanotechnology, which involves the design, characterization, production, and application of nanoscale drug delivery systems. In medicine, nanotechnologies, such as polymeric nanoparticles, solid lipid nanoparticles, nanostructured lipid carriers, gold nanoparticles, hydrogels, cyclodextrin complexes, and liquid crystals, are promising tools for diagnostic probes and therapeutic devices. The objective of this study is to present a systematic review of nanotechnology-based drug delivery systems for oral cancers.

  4. Nano-carrier based drug delivery systems for sustained antimicrobial agent release from orthopaedic cementous material.

    Science.gov (United States)

    Al Thaher, Yazan; Perni, Stefano; Prokopovich, Polina

    2017-11-01

    Total joint replacement (TJR), such as hip and knee replacement, is a popular procedure worldwide. Prosthetic joint infections (PJI) after this procedure have been widely reported, where treatment of such infections is complex with high cost and prolonged hospital stay. In cemented arthroplasties, the use of antibiotic loaded bone cement (ALBC) is a standard practice for the prophylaxis and treatment of PJI. Recently, the development of bacterial resistance by pathogenic microorganisms against most commonly used antibiotics increased the interest in alternative approaches for antimicrobial delivery systems such as nanotechnology. This review summarizes the efforts made to improve the antimicrobial properties of PMMA bone cements using nanotechnology based antibiotic and non-antibiotic delivery systems to overcome drawbacks of ALBC in the prophylaxis and treatment of PJIs after hip and knee replacement. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Recent Advances on Cellulose-Based Nano-Drug Delivery Systems: Design of Prodrugs and Nanoparticles.

    Science.gov (United States)

    Dai, Lin; Si, Chuanling

    2017-07-11

    Cellulose being the first abundant biopolymers in nature is playing an increasing role as a carrier material in the biomedicine field. Compared with other materials, cellulose has its own advantages, including rich in resources, good thermal and chemical stability, high hydrophilicity, and excellent biocompatibility, etc. These fascinating properties have made cellulose a real potential material to create nano-drug delivery systems. However, the structure of pure cellulose caused some inherent shortcomings, for instance, bad plasticity and dimensional stability, poor solubility in ordinary solvents, and lack of antibacterial activity. Therefore, it is indispensable to modify the morphology and microstructure of cellulose. This review presents some remarkable recent advances on the drug delivery applications of cellulose-based prodrugs and nanoparticles. Various methods employed for the functionalization, pharmacodynamic actions and applications are covered. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  6. Intranasal delivery of nanoparticle-based vaccine increases protection against S. pneumoniae

    Energy Technology Data Exchange (ETDEWEB)

    Mott, Brittney [University of North Texas Health Science Center, Department of Molecular Biology and Immunology (United States); Thamake, Sanjay [Radio-Isotope Therapy of America Foundation (United States); Vishwanatha, Jamboor; Jones, Harlan P., E-mail: harlan.jones@unthsc.edu [University of North Texas Health Science Center, Department of Molecular Biology and Immunology (United States)

    2013-05-15

    Nanoparticle (NP) technologies are becoming commonplace in the development of vaccine delivery systems to protect against various diseases. The current study determined the efficacy of intranasal delivery of a 234 {+-} 87.5 nm poly lactic-co-glycolic acid nanoparticle vaccine construct in establishing protection against experimental respiratory pneumococcal infection. Nanoparticles encapsulating heat-killed Streptococcus pneumoniae (NP-HKSP) were retained in the lungs 11 days following nasal administration compared to empty NP. Immunization with NP-HKSP produced significant resistance against S. pneumoniae infection compared to administration of HKSP alone. Increased protection correlated with a significant increase in antigen-specific Th1-associated IFN-{gamma} cytokine response by pulmonary lymphocytes. This study establishes the efficacy of NP-based technology as a non-invasive and targeted approach for nasal-pulmonary immunization against pulmonary infections.

  7. Nanotechnology-Based Drug Delivery Systems for Melanoma Antitumoral Therapy: A Review

    Directory of Open Access Journals (Sweden)

    Roberta Balansin Rigon

    2015-01-01

    Full Text Available Melanoma (MEL is a less common type of skin cancer, but it is more aggressive with a high mortality rate. The World Cancer Research Fund International (GLOBOCAN 2012 estimates that there were 230,000 new cases of MEL in the world in 2012. Conventional MEL treatment includes surgery and chemotherapy, but many of the chemotherapeutic agents used present undesirable properties. Drug delivery systems are an alternative strategy by which to carry antineoplastic agents. Encapsulated drugs are advantageous due to such properties as high stability, better bioavailability, controlled drug release, a long blood circulation time, selective organ or tissue distribution, a lower total required dose, and minimal toxic side effects. This review of scientific research supports applying a nanotechnology-based drug delivery system for MEL therapy.

  8. Montmorillonite Clay-Based Polyurethane Nanocomposite As Local Triamcinolone Acetonide Delivery System

    Directory of Open Access Journals (Sweden)

    Flávia Carmo Horta Pinto

    2011-01-01

    Full Text Available Biodegradable polyurethane was synthesized by preparing aqueous polyurethane dispersion having poly(caprolactone and poly(ethylene glycol as soft segments. Montmorillonite particles were delaminated within the waterborne polyurethane to produce a nanocomposite. The triamcinolone acetonide (TA, an important corticoid drug, was dispersed into the nanocomposite followed by a drying step to produce an implantable drug delivery system. Infrared (FTIR results demonstrated that the original chemical structure of the TA was preserved after incorporation into the nanocomposite. Wide angle (WAXS and small angle X-ray scattering (SAXS results suggested that TA and clay do not dramatically change the morphology phase of the polymer although they can interact with each other. The presence of montmorillonite particles in the nanocomposite reduced the rate of TA release as compared to the pure polyurethane and enhanced the mechanical properties of the polymer. The overall results indicate that montmorillonite clay-based polyurethane nanocomposite could be potentially applied as local TA delivery system.

  9. The application of polysaccharide-based nanogels in peptides/proteins and anticancer drugs delivery.

    Science.gov (United States)

    Zhang, Lin; Pan, Jifei; Dong, Shibo; Li, Zhaoming

    2017-09-01

    Finding adequate carriers for proteins/peptides and anticancer drugs delivery has become an urgent need, owing to the growing number of therapeutic macromolecules and the increasing amount of cancer incidence. Polysaccharide-based nanogels have attracted interest as carriers for proteins/peptides and anticancer drugs because of their characteristic properties like biodegradability, biocompatibility, stimuli-responsive behaviour, softness and swelling to help achieve a controlled, triggered response at the target site. In addition, the groups of the polysaccharide backbone are able to be modified to develop functional nanogels. Some polysaccharides have the intrinsic ability to recognise specific cell types, allowing the design of targeted drug delivery systems through receptor-mediated endocytosis. This review is aimed at describing and exploring the potential of polysaccharides that are used in nanogels which can help to deliver proteins/peptides and anticancer drugs.

  10. Versatile types of polysaccharide-based supramolecular polycation/pDNA nanoplexes for gene delivery

    Science.gov (United States)

    Hu, Yang; Zhao, Nana; Yu, Bingran; Liu, Fusheng; Xu, Fu-Jian

    2014-06-01

    Different polysaccharide-based supramolecular polycations were readily synthesized by assembling multiple β-cyclodextrin-cored star polycations with an adamantane-functionalized dextran via host-guest interaction in the absence or presence of bioreducible linkages. Compared with nanoplexes of the starting star polycation and pDNA, the supramolecular polycation/pDNA nanoplexes exhibited similarly low cytotoxicity, improved cellular internalization and significantly higher gene transfection efficiencies. The incorporation of disulfide linkages imparted the supramolecular polycation/pDNA nanoplexes with the advantage of intracellular bioreducibility, resulting in better gene delivery properties. In addition, the antitumor properties of supramolecular polycation/pDNA nanoplexes were also investigated using a suicide gene therapy system. The present study demonstrates that the proper assembly of cyclodextrin-cored polycations with adamantane-functionalized polysaccharides is an effective strategy for the production of new nanoplex delivery systems.Different polysaccharide-based supramolecular polycations were readily synthesized by assembling multiple β-cyclodextrin-cored star polycations with an adamantane-functionalized dextran via host-guest interaction in the absence or presence of bioreducible linkages. Compared with nanoplexes of the starting star polycation and pDNA, the supramolecular polycation/pDNA nanoplexes exhibited similarly low cytotoxicity, improved cellular internalization and significantly higher gene transfection efficiencies. The incorporation of disulfide linkages imparted the supramolecular polycation/pDNA nanoplexes with the advantage of intracellular bioreducibility, resulting in better gene delivery properties. In addition, the antitumor properties of supramolecular polycation/pDNA nanoplexes were also investigated using a suicide gene therapy system. The present study demonstrates that the proper assembly of cyclodextrin-cored polycations

  11. Exploring the Applicability of Nano-Poration for Remote Control in Smart Drug Delivery Systems.

    Science.gov (United States)

    Denzi, Agnese; Della Valle, Elena; Apollonio, Francesca; Breton, Marie; Mir, Lluis M; Liberti, Micaela

    2017-02-01

    Smart drug delivery systems represent an interesting tool to significantly improve the efficiency and the precision in the treatment of a broad category of diseases. In this context, a drug delivery mediated by nanosecond pulsed electric fields seems a promising technique, allowing for a controlled release and uptake of drugs by the synergy between the electropulsation and nanocarriers with encapsulated drugs. The main concern about the use of electroporation for drug delivery applications is the difference in dimension between the liposome (nanometer range) and the cell (micrometer range). The choice of liposome dimension is not trivial. Liposomes larger than 500 nm of diameter could be recognized as pathogen agents by the immune system, while liposomes of smaller size would require external electric field of high amplitudes for the membrane electroporation that could compromise the cell viability. The aim of this work is to theoretically study the possibility of a simultaneous cell and liposomes electroporation. The numerical simulations reported the possibility to electroporate the cell and a significant percentage of liposomes with comparable values of external electric field, when a 12 nsPEF is used.

  12. Studies on mRNA electroporation of immature and mature dendritic cells

    DEFF Research Database (Denmark)

    Met, Ozcan; Eriksen, Jens; Svane, Inge Marie

    2008-01-01

    Previous studies have shown that mRNA-electroporated dendritic cells (DCs) are able to process and present tumor-associated antigens, leading to the activation of tumor-specific T cells in vitro and in vivo. However, the optimal maturation state of antigen loading and half-life of the m...... in addition to higher T-cell stimulatory ability compared to transfection of DCs prior to maturation. Mature mRNA-electroporated DCs showed long-lived expression of EGFP and were able to stimulate influenza matrix protein M1 (M1)-specific T cells up to 24 h after electroporation. However, when DCs were...... subjected to increasing electrical pulses the level of transgene expression was four-fold upregulated, equipping these DCs to be more potent in inducing M1-specific T cells. Also, the application of long electrical pulses induced further upregulation of HLA-DR, CD80, and CD86 expression in mature DCs...

  13. Electroporation induced by internal defibrillation shock with and without recovery in intact rabbit hearts

    Science.gov (United States)

    Wang, Yves T.; Efimov, Igor R.

    2012-01-01

    Defibrillation shocks from implantable cardioverter defibrillators can be lifesaving but can also damage cardiac tissues via electroporation. This study characterizes the spatial distribution and extent of defibrillation shock-induced electroporation with and without a 45-min postshock period for cell membranes to recover. Langendorff-perfused rabbit hearts (n = 31) with and without a chronic left ventricular (LV) myocardial infarction (MI) were studied. Mean defibrillation threshold (DFT) was determined to be 161.4 ± 17.1 V and 1.65 ± 0.44 J in MI hearts for internally delivered 8-ms monophasic truncated exponential (MTE) shocks during sustained ventricular fibrillation (>20 s, SVF). A single 300-V MTE shock (twice determined DFT voltage) was used to terminate SVF. Shock-induced electroporation was assessed by propidium iodide (PI) uptake. Ventricular PI staining was quantified by fluorescent imaging. Histological analysis was performed using Masson's Trichrome staining. Results showed PI staining concentrated near the shock electrode in all hearts. Without recovery, PI staining was similar between normal and MI groups around the shock electrode and over the whole ventricles. However, MI hearts had greater total PI uptake in anterior (P < 0.01) and posterior (P < 0.01) LV epicardial regions. Postrecovery, PI staining was reduced substantially, but residual staining remained significant with similar spacial distributions. PI staining under SVF was similar to previously studied paced hearts. In conclusion, electroporation was spatially correlated with the active region of the shock electrode. Additional electroporation occurred in the LV epicardium of MI hearts, in the infarct border zone. Recovery of membrane integrity postelectroporation is likely a prolonged process. Short periods of SVF did not affect electroporation injury. PMID:22730387

  14. Exercise during pregnancy and risk of cesarean delivery in nulliparous women: a large population-based cohort study.

    Science.gov (United States)

    Owe, Katrine Mari; Nystad, Wenche; Stigum, Hein; Vangen, Siri; Bø, Kari

    2016-12-01

    Vaginal delivery for the first birth is of great importance for further obstetric performance for the individual woman. Given the rising cesarean delivery rates worldwide over the past decades, a search for modifiable factors that are associated with cesarean delivery is needed. Exercise may be a modifiable factor that is associated with type of delivery, but the results of previous studies are not conclusive. The purpose of this study was to investigate the association between exercise during pregnancy and cesarean delivery, both acute and elective, in nulliparous women. We conducted a population-based cohort study that involved 39,187 nulliparous women with a singleton pregnancy who were enrolled in the Norwegian Mother and Child Cohort Study between 2000 and 2009. All women answered 2 questionnaires in pregnancy weeks 17 and 30. Acute and elective cesarean delivery data were obtained from the Medical Birth Registry of Norway. Information on exercise frequency and type was assessed prospectively by questionnaires in pregnancy weeks 17 and 30. Generalized linear models estimated risk differences of acute and elective cesarean delivery for different frequencies and types of exercise during pregnancy weeks 17 and 30. We used restricted cubic splines to examine dose-response associations of exercise frequency and acute cesarean delivery. A test for nonlinearity was also conducted. The total cesarean delivery rate was 15.4% (n=6030), of which 77.8% (n=4689) was acute cesarean delivery. Exercise during pregnancy was associated with a reduced risk of cesarean delivery, particularly for acute cesarean delivery. A nonlinear association was observed for exercise frequency in weeks 17 and 30 and risk of acute cesarean delivery (test for nonlinearity, P=.003 and P=.027, respectively). The largest risk reduction was observed for acute cesarean delivery among women who exercised >5 times weekly during weeks 17 (-2.2%) and 30 (-3.6%) compared with nonexercisers (test for trend

  15. Tobacco mosaic virus-based protein nanoparticles and nanorods for chemotherapy delivery targeting breast cancer

    Science.gov (United States)

    Bruckman, Michael A.; Czapar, Anna E.; VanMeter, Allen; Randolph, Lauren N.; Steinmetz, Nicole F.

    2016-01-01

    Drug delivery systems are required for drug targeting to avoid adverse effects associated with chemotherapy treatment regimes. Our approach is focused on the study and development of plant virus-based materials as drug delivery systems; specifically, this work focuses on the tobacco mosaic virus (TMV). Native TMV forms a hollow, high aspect-ratio nanotube measuring 300 × 18 nm with a 4 nm-wide central channel. Heat-transformation can be applied to TMV yielding spherical nanoparticles (SNPs) measuring ~50 nm in size. While bioconjugate chemistries have been established to modify the TMV rod, such methods have not yet been described for the SNP platform. In this work, we probed the reactivity of SNPs toward bioconjugate reactions targeting lysine, glutamine/aspartic acid, and cysteine residues. We demonstrate functionalization of SNPs using these chemistries yielding efficient payload conjugation. In addition to covalent labeling techniques, we developed encapsulation techniques, where the cargo is loaded into the SNP during heat-transition from rod-to-sphere. Finally, we developed TMV and SNP formulations loaded with the chemotherapeutic doxorubicin, and we demonstrate the application of TMV rods and spheres for chemotherapy delivery targeting breast cancer. PMID:26941034

  16. Biodegradable stereocomplex micelles based on dextran-block-polylactide as efficient drug deliveries.

    Science.gov (United States)

    Zhao, Ziwei; Zhang, Zhe; Chen, Li; Cao, Yue; He, Chaoliang; Chen, Xuesi

    2013-10-22

    Biodegradable stereocomplex micelles (SCMs) based on amphiphilic dextran-block-polylactide (Dex-b-PLA) were designed and used for efficient intracellular drug deliveries. The Dex-b-PLA copolymers were successfully synthesized by click reaction. The structures of the resultant copolymers were verified by (1)H NMR and FT-IR spectra. The formation of stable micelles through self-assembly driven by the stereocomplexation between enantiomeric l- and d-PLA blocks was characterized by transmission electron microscopy (TEM), dynamic laser scattering (DLS), and fluorescence techniques. It was interesting to observe that the SCMs showed lower critical micelle concentration values (CMCs) because of the stereocomplex interaction between PLLA and PDLA. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis provided information on the thermal and crystal properties of the copolymers and SCMs. The improved stability of SCMs should be attractive for intracellular drug delivery. Thus, a model anticancer drug doxorubicin (DOX) was loaded into micelles, and the in vitro drug release in was also studied. The release kinetics of DOX showed DOX-loaded SCMs exhibited slower DOX release. Confocal laser scanning microscopy (CLSM) and flow cytometry studies also showed that the DOX-loaded SCMs exhibited a slower drug release behavior. Meanwhile, the MTT assay demonstrated that DOX-loaded SCMs show lower cellular proliferation inhibition against HepG2. In sum, the micelles through self-assembly driven by stereocomplex interaction would have great potential to be used as stable delivery vehicles for pharmaceutical and biomedical applications.

  17. Painless drug delivery through microneedle-based transdermal patches featuring active infusion.

    Science.gov (United States)

    Roxhed, Niclas; Samel, Björn; Nordquist, Lina; Griss, Patrick; Stemme, Göran

    2008-03-01

    This paper presents the first microneedle-based transdermal patch with integrated active dispensing functionality. The electrically controlled system consists of a low-cost dosing and actuation unit capable of controlled release of liquid in the microliter range at low flow-rates and minimally invasive, side-opened, microneedles. The system was successfully tested in vivo by insulin administration to diabetic rats. Active infusion of insulin at 2 mul/h was compared to passive, diffusion-driven, delivery. Continuous active infusion caused significantly higher insulin concentrations in blood plasma. After a 3-h delivery period, the insulin concentration was five times larger compared to passive delivery. Consistent with insulin concentrations, actively administered insulin resulted in a significant decrease of blood glucose levels. Additionally, insertion and liquid injection was verified on human skin. This study shows the feasibility of a patch-like system with on-board liquid storage and dispensing capability. The proposed device represents a first step towards painless and convenient administration of macromolecular drugs such as insulin or vaccines.

  18. Drug-Delivery System Based on Salmon DNA Nano- and Micro-Scale Structures.

    Science.gov (United States)

    Lee, Yunwoo; Dugansani, Sreekantha Reddy; Jeon, So Hee; Hwang, Soon Hyoung; Kim, Jae-Hyun; Park, Sung Ha; Jeong, Jun-Ho

    2017-08-29

    Microneedles, fabricated by nano-moulding technology show great promise in the field of drug delivery by enabling the painless self-administration of drugs in a patient-friendly manner. In this study, double-stranded salmon DNA (SDNA) was used as both a drug-delivery vehicle and structural material with a microneedle system. SDNA is non-toxic and demonstrates good mechanical robustness, mouldability, biocompatibility, bio-absorbability, and binding affinity with drug molecules for bio-functional applications. Benign fabrication conditions to protect temperature-sensitive biomolecules are used to produce SDNA structures of various sizes with a high aspect ratio (4: 1). Unlike existing dissolving microneedle structure materials, the special binding characteristics of doxorubicin hydrochloride, anti-cancer drug molecules, and SDNA demonstrate the stability of drug-molecule encapsulation via UV-absorption and photoluminescence analyses. Based on COMSOL simulation and in vitro analysis of the stratum corneum of porcine skin, the mechanical functionality of SDNA microneedles was evaluated in vitro by penetrating the stratum corneum of porcine skin. The SDNA microneedle dissolved and drug permeation was assessed using rhodamine, a drug surrogate. Owing to its many beneficial characteristics, we anticipate that the SDNA microneedle platform will serve as an effective alternative for drug delivery.

  19. Drug delivery via lipoprotein-based carriers: answering the challenges in systemic therapeutics.

    Science.gov (United States)

    Sabnis, Nirupama; Lacko, Andras G

    2012-05-01

    Plasma lipoproteins are transporters of lipids and other hydrophobic molecules in the mammalian circulation. Lipoproteins also have a strong potential to serve as drug-delivery vehicles due to their small size, long residence time in the circulation and high-drug payload. Consequently, lipoproteins and synthetic/reconstituted lipoprotein preparations have been evaluated with increasing interest towards clinical applications, particularly for cancer diagnostics/imaging and chemotherapy. In this review, past and current studies on lipoproteins and similar alternative drug carriers are discussed regarding their suitability as agents to deliver drugs, primarily to cancer cells and tumors. A lipoprotein-based delivery strategy may also provide a novel platform for improving the therapeutic efficacy of drugs that have previously been judged unsuitable or had only limited application due to poor solubility. An additional, and perhaps the most important aspect of the drug-delivery process via lipoprotein-type carriers, is the receptor-mediated uptake of the payload from the lipoprotein complex. Monitoring the expression of specific receptors prior to treatment could, thus, give rise to efficient selection of optimally responsive patients, resulting in a successful personalized therapy regimen.

  20. Biopolymer-Based Nanoparticles for Drug/Gene Delivery and Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Keiji Numata

    2013-01-01

    Full Text Available There has been a great interest in application of nanoparticles as biomaterials for delivery of therapeutic molecules such as drugs and genes, and for tissue engineering. In particular, biopolymers are suitable materials as nanoparticles for clinical application due to their versatile traits, including biocompatibility, biodegradability and low immunogenicity. Biopolymers are polymers that are produced from living organisms, which are classified in three groups: polysaccharides, proteins and nucleic acids. It is important to control particle size, charge, morphology of surface and release rate of loaded molecules to use biopolymer-based nanoparticles as drug/gene delivery carriers. To obtain a nano-carrier for therapeutic purposes, a variety of materials and preparation process has been attempted. This review focuses on fabrication of biocompatible nanoparticles consisting of biopolymers such as protein (silk, collagen, gelatin, β-casein, zein and albumin, protein-mimicked polypeptides and polysaccharides (chitosan, alginate, pullulan, starch and heparin. The effects of the nature of the materials and the fabrication process on the characteristics of the nanoparticles are described. In addition, their application as delivery carriers of therapeutic drugs and genes and biomaterials for tissue engineering are also reviewed.

  1. Potential for Layered Double Hydroxides-Based, Innovative Drug Delivery Systems

    Directory of Open Access Journals (Sweden)

    Kai Zhang

    2014-04-01

    Full Text Available Layered Double Hydroxides (LDHs-based drug delivery systems have, for many years, shown great promises for the delivery of chemical therapeutics and bioactive molecules to mammalian cells in vitro and in vivo. This system offers high efficiency and drug loading density, as well as excellent protection of loaded molecules from undesired degradation. Toxicological studies have also found LDHs to be biocompatible compared with other widely used nanoparticles, such as iron oxide, silica, and single-walled carbon nanotubes. A plethora of bio-molecules have been reported to either attach to the surface of or intercalate into LDH materials through co-precipitation or anion-exchange reaction, including amino acid and peptides, ATPs, vitamins, and even polysaccharides. Recently, LDHs have been used for gene delivery of small molecular nucleic acids, such as antisense, oligonucleotides, PCR fragments, siRNA molecules or sheared genomic DNA. These nano-medicines have been applied to target cells or organs in gene therapeutic approaches. This review summarizes current progress of the development of LDHs nanoparticle drug carriers for nucleotides, anti-inflammatory, anti-cancer drugs and recent LDH application in medical research. Ground breaking studies will be highlighted and an outlook of the possible future progress proposed. It is hoped that the layered inorganic material will open up new frontier of research, leading to new nano-drugs in clinical applications.

  2. Polysaccharide based nanogels in the drug delivery system: Application as the carrier of pharmaceutical agents.

    Science.gov (United States)

    Debele, Tilahun Ayane; Mekuria, Shewaye Lakew; Tsai, Hsieh-Chih

    2016-11-01

    Polysaccharide-based nanoparticles have fascinated attention as a vesicle of different pharmaceutical agents due to their unique multi-functional groups in addition to their physicochemical properties, including biocompatibility and biodegradability. The existence of multi-functional groups on the polysaccharide backbone permits facile chemical or biochemical modification to synthesize polysaccharide based nanoparticles with miscellaneous structures. Polysaccharide-based nanogels have high water content, large surface area for multivalent bioconjugation, tunable size, and interior network for the incorporation of different pharmaceutical agents. These unique properties offer great potential for the utilization of polysaccharide-based nanogels in the drug delivery systems. Hence, this review describes chemistry of certain common polysaccharides, several methodologies used to synthesize polysaccharide nanoparticles and primarily focused on the polysaccharide (or polysaccharide derivative) based nanogels as the carrier of pharmaceutical agents. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Changes in optical properties of electroporated cells as revealed by digital holographic microscopy.

    Science.gov (United States)

    Calin, Violeta L; Mihailescu, Mona; Mihale, Nicolae; Baluta, Alexandra V; Kovacs, Eugenia; Savopol, Tudor; Moisescu, Mihaela G

    2017-04-01

    Changes in optical and shape-related characteristics of B16F10 cells after electroporation were investigated using digital holographic microscopy (DHM). Bipolar rectangular pulses specific for electrochemotherapy were used. Electroporation was performed in an "off-axis" DHM set-up without using exogenous markers. Two types of cell parameters were monitored seconds and minutes after pulse train application: parameters addressing a specifically defined area of the cell (refractive index and cell height) and global cell parameters (projected area, optical phase shift profile and dry mass). The biphasic behavior of cellular parameters was explained by water and mannitol dynamics through the electropermeabilized cell membrane.

  4. Changes in optical properties of electroporated cells as revealed by digital holographic microscopy

    Science.gov (United States)

    Calin, Violeta L.; Mihailescu, Mona; Mihale, Nicolae; Baluta, Alexandra V.; Kovacs, Eugenia; Savopol, Tudor; Moisescu, Mihaela G.

    2017-01-01

    Changes in optical and shape-related characteristics of B16F10 cells after electroporation were investigated using digital holographic microscopy (DHM). Bipolar rectangular pulses specific for electrochemotherapy were used. Electroporation was performed in an “off-axis” DHM set-up without using exogenous markers. Two types of cell parameters were monitored seconds and minutes after pulse train application: parameters addressing a specifically defined area of the cell (refractive index and cell height) and global cell parameters (projected area, optical phase shift profile and dry mass). The biphasic behavior of cellular parameters was explained by water and mannitol dynamics through the electropermeabilized cell membrane. PMID:28736667

  5. Nanobody-Based Delivery Systems for Diagnosis and Targeted Tumor Therapy

    Directory of Open Access Journals (Sweden)

    Yaozhong Hu

    2017-11-01

    Full Text Available The development of innovative targeted therapeutic approaches are expected to surpass the efficacy of current forms of treatments and cause less damage to healthy cells surrounding the tumor site. Since the first development of targeting agents from hybridoma’s, monoclonal antibodies (mAbs have been employed to inhibit tumor growth and proliferation directly or to deliver effector molecules to tumor cells. However, the full potential of such a delivery strategy is hampered by the size of mAbs, which will obstruct the targeted delivery system to access the tumor tissue. By serendipity, a new kind of functional homodimeric antibody format was discovered in camelidae, known as heavy-chain antibodies (HCAbs. The cloning of the variable domain of HCAbs produces an attractive minimal-sized alternative for mAbs, referred to as VHH or nanobodies (Nbs. Apart from their dimensions in the single digit nanometer range, the unique characteristics of Nbs combine a high stability and solubility, low immunogenicity and excellent affinity and specificity against all possible targets including tumor markers. This stimulated the development of tumor-targeted therapeutic strategies. Some autonomous Nbs have been shown to act as antagonistic drugs, but more importantly, the targeting capacity of Nbs has been exploited to create drug delivery systems. Obviously, Nb-based targeted cancer therapy is mainly focused toward extracellular tumor markers, since the membrane barrier prevents antibodies to reach the most promising intracellular tumor markers. Potential strategies, such as lentiviral vectors and bacterial type 3 secretion system, are proposed to deliver target-specific Nbs into tumor cells and to block tumor markers intracellularly. Simultaneously, Nbs have also been employed for in vivo molecular imaging to diagnose diseased tissues and to monitor the treatment effects. Here, we review the state of the art and focus on recent developments with Nbs as

  6. Improved chitosan-mediated gene delivery based on easily dissociated chitosan polyplexes of highly defined chitosan oligomers

    National Research Council Canada - National Science Library

    Köping-Höggård, M; Vårum, K M; Issa, M; Danielsen, S; Christensen, B E; Stokke, B T; Artursson, P

    2004-01-01

    Nonviral gene delivery systems based on conventional high-molecular-weight chitosans are efficient after lung administration in vivo, but have poor physical properties such as aggregated shapes, low...

  7. Location-based health information services: a new paradigm in personalised information delivery

    Directory of Open Access Journals (Sweden)

    Boulos Maged

    2003-01-01

    Full Text Available Abstract Brute health information delivery to various devices can be easily achieved these days, making health information instantly available whenever it is needed and nearly anywhere. However, brute health information delivery risks overloading users with unnecessary information that does not answer their actual needs, and might even act as noise, masking any other useful and relevant information delivered with it. Users' profiles and needs are definitely affected by where they are, and this should be taken into consideration when personalising and delivering information to users in different locations. The main goal of location-based health information services is to allow better presentation of the distribution of health and healthcare needs and Internet resources answering them across a geographical area, with the aim to provide users with better support for informed decision-making. Personalised information delivery requires the acquisition of high quality metadata about not only information resources, but also information service users, their geographical location and their devices. Throughout this review, experience from a related online health information service, HealthCyberMap http://healthcybermap.semanticweb.org/, is referred to as a model that can be easily adapted to other similar services. HealthCyberMap is a Web-based directory service of medical/health Internet resources exploring new means to organise and present these resources based on consumer and provider locations, as well as the geographical coverage or scope of indexed resources. The paper also provides a concise review of location-based services, technologies for detecting user location (including IP geolocation, and their potential applications in health and healthcare.

  8. Nanobiotechnology-based delivery strategies: New frontiers in brain tumor targeted therapies.

    Science.gov (United States)

    Mangraviti, Antonella; Gullotti, David; Tyler, Betty; Brem, Henry

    2016-10-28

    Despite recent technological advancements and promising preclinical experiments, brain tumor patients are still met with limited treatment options. Some of the barriers to clinical improvements include the systemic toxicity of cytotoxic compounds, the impedance of the blood brain barrier (BBB), and the lack of therapeutic agents that can selectively target the intracranial tumor environment. To overcome such barriers, a number of chemotherapeutic agents and nucleic acid-based therapies are rapidly being synthesized and tested as new brain tumor-targeted delivery strategies. Novel carriers include liposomal and polymeric nanoparticles, wafers, microchips, microparticle-based nanoplatforms and cells-based vectors. Strong preclinical results suggest that these nanotechnologies are set to transform the therapeutic paradigm for brain tumor treatment. In addition to new tumoricidal agents, parallel work is also being conducted on the BBB front. Preclinical testing of chemical and physical modulation strategies is yielding improved intracranial concentrations. New diagnostic and therapeutic imaging techniques, such as high-intensity focused ultrasound and MRI-guided focused ultrasound, are being used to modulate the BBB in a more precise and non-invasive manner. This review details some of the tremendous advances that are being explored in current brain tumor targeted therapies, including local implant development, nanobiotechnology-based delivery strategies, and techniques of BBB manipulation. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Potentials of Chitosan-Based Delivery Systems in Wound Therapy: Bioadhesion Study

    Directory of Open Access Journals (Sweden)

    Julia Hurler

    2012-01-01

    Full Text Available Chitosan is currently proposed to be one of the most promising polymers in wound dressing development. Our research focuses on its potential as a vehicle for nano-delivery systems destined for burn therapy. One of the most important features of wound dressing is its bioadhesion to the wounded site. We compared the bioadhesive properties of chitosan with those of Carbopol, a synthetic origin polymer. Chitosan-based hydrogels of different molecular weights were first analyzed by texture analysis for gel cohesiveness, adhesiveness and hardness. In vitro release studies showed no difference in release of model antimicrobial drug from the different hydrogel formulations. Bioadhesion tests were performed on pig ear skin and the detachment force, necessary to remove the die from the skin, and the amount of remaining formulation on the skin were determined. Although no significant difference regarding detachment force could be seen between Carbopol-based and chitosan-based formulations, almost double the amount of chitosan formulation remained on the skin as compared to Carbopol formulations. The findings confirmed the great potential of chitosan-based delivery systems in advanced wound therapy. Moreover, results suggest that formulation retention on the ex vivo skin samples could provide deeper insight on formulation bioadhesiveness than the determination of detachment force.

  10. Transtheoretical Model-based Nursing Intervention on Lifestyle Change: A Review Focused on Intervention Delivery Methods.

    Science.gov (United States)

    Lee, Joo Yun; Park, Hyeoun-Ae; Min, Yul Ha

    2015-06-01

    The transtheoretical model (TTM) was used to provide tailored nursing for lifestyle management such as diet, physical activity, and smoking cessation. The present study aims to assess the provision of intervention delivery methods, intervention elements, and stage-matched interventions, in order to identify ways in which information technology is used in the TTM-based research. The relevant literature was selected by two researchers using inclusion criteria after searching for "TTM (transtheoretical or stage of change)" and "nursing" from the databases PubMed and CINAHL. The selected studies were categorized in terms of study characteristics, intervention delivery method, intervention element, and use and level of stage-matched intervention. A total of 35 studies were selected including eight studies that used information communication technology (ICT). Nine different intervention delivery methods were used, of which "face-to-face" was the most common at 24 times. Of the 35 studies, 26 provided stage-matched interventions. Seven different intervention elements were used, of which "counseling" was the most common at 27 times. Of all the intervention elements, tailored feedback used ICT the most at seven instances out of nine, and there was a significant difference in the rate of ICT usage among intervention elements. ICT is not yet actively used in the TTM-based nursing interventions. Stage-matched interventions and TTM concepts were shown to be in partial use also in the TTM-based interventions. Therefore, it is necessary to develop a variety of ways to use ICT in tailored nursing interventions and to use TTM frameworks and concepts. Copyright © 2015. Published by Elsevier B.V.

  11. Human artificial chromosome-based gene delivery vectors for biomedicine and biotechnology.

    Science.gov (United States)

    Kouprina, Natalay; Tomilin, Alexey N; Masumoto, Hiroshi; Earnshaw, William C; Larionov, Vladimir

    2014-04-01

    Human artificial chromosomes (HACs) have several advantages over viruses as gene delivery vectors, including stable episomal maintenance in a single copy and the ability to carry large gene inserts. In this review, we summarise recent work on gene transfer into mammalian cells using the HACs. HACs allow therapeutic transgenes to be expressed in target cells under conditions that recapitulate the physiological regulation of endogenous loci. Based on the published data, the HAC vectors have a great potential for gene therapy, regenerative medicine, screening of anticancer drugs and biotechnology.

  12. Passivity-Based Automated Design of Stable Multi-Feedback Distributed Power Delivery Systems

    Science.gov (United States)

    2017-03-01

    performan pically determ C gain, phase stributed on- delivering hig ce systems. S n automated delivery is pr is designe fabricated in performance a n-chip...termined by t In systems w igure 1, the se no straig table loop. An een proposed the power g design proces this work b on with a pa iques...Performan ce of an i ined by a se margin, DC o chip power h quality pow tability is a pr design flow f oposed. A dis d based on 28 nm CMO nd stable

  13. MRI-based treatment planning and dose delivery verification for intraocular melanoma brachytherapy.

    Science.gov (United States)

    Zoberi, Jacqueline Esthappan; Garcia-Ramirez, Jose; Hedrick, Samantha; Rodriguez, Vivian; Bertelsman, Carol G; Mackey, Stacie; Hu, Yanle; Gach, H Michael; Rao, P Kumar; Grigsby, Perry W

    2017-08-14

    Episcleral plaque brachytherapy (EPB) planning is conventionally based on approximations of the implant geometry with no volumetric imaging following plaque implantation. We have developed an MRI-based technique for EPB treatment planning and dose delivery verification based on the actual patient-specific geometry. MR images of 6 patients, prescribed 85 Gy over 96 hours from Collaborative Ocular Melanoma Study-based EPB, were acquired before and after implantation. Preimplant and postimplant scans were used to generate "preplans" and "postplans", respectively. In the preplans, a digital plaque model was positioned relative to the tumor, sclera, and nerve. In the postplans, the same plaque model was positioned based on the imaged plaque. Plaque position, point doses, percentage of tumor volume receiving 85 Gy (V100), and dose to 100% of tumor volume (Dmin) were compared between preplans and postplans. All isodose plans were computed using TG-43 formalism with no heterogeneity corrections. Shifts and tilts of the plaque ranged from 1.4 to 8.6 mm and 1.0 to 3.8 mm, respectively. V100 was ≥97% for 4 patients. Dmin for preplans and postplans ranged from 83 to 118 Gy and 45 to 110 Gy, respectively. Point doses for tumor apex and base were all found to decrease from the preimplant to the postimplant plan, with mean differences of 16.7 ± 8.6% and 30.5 ± 11.3%, respectively. By implementing MRI for EPB, we eliminate reliance on approximations of the eye and tumor shape and the assumption of idealized plaque placement. With MRI, one can perform preimplant as well as postimplant imaging, facilitating EPB treatment planning based on the actual patient-specific geometry and dose-delivery verification based on the imaged plaque position. Copyright © 2017 American Brachytherapy Society. Published by Elsevier Inc. All rights reserved.

  14. [Implementation methodology of practices based on scientific evidence for assistance in natural delivery: a pilot study].

    Science.gov (United States)

    Côrtes, Clodoaldo Tentes; Santos, Rafael Cleison Silva Dos; Caroci, Adriana de Souza; Oliveira, Sheyla Guimarães; Oliveira, Sonia Maria Junqueira Vasconcelos de; Riesco, Maria Luiza Gonzalez

    2015-10-01

    Presenting methodology for transferring knowledge to improve maternal outcomes in natural delivery based on scientific evidence. An intervention study conducted in the maternity hospital of Itapecerica da Serra, SP, with 50 puerperal women and 102 medical records from July to November 2014. The PACES tool from Joanna Briggs Institute, consisting of pre-clinical audit (phase 1), implementation of best practice (phase 2) and Follow-up Clinical Audit (phase 3) was used. Data were analyzed by comparing results of phases 1 and 3 with Fisher's exact test and a significance level of 5%. The vertical position was adopted by the majority of puerperal women with statistical difference between phases 1 and 3. A significant increase in bathing/showering, walking and massages for pain relief was found from the medical records. No statistical difference was found in other practices and outcomes. Barriers and difficulties in the implementation of evidence-based practices have been identified. Variables were refined, techniques and data collection instruments were verified, and an intervention proposal was made. The study found possibilities for implementing a methodology of practices based on scientific evidence for assistance in natural delivery.

  15. Aminated carbon-based "cargo vehicles" for improved delivery of methotrexate to breast cancer cells.

    Science.gov (United States)

    Joshi, Mayank; Kumar, Pramod; Kumar, Rajendra; Sharma, Gajanand; Singh, Bhupinder; Katare, Om Prakash; Raza, Kaisar

    2017-06-01

    The purpose of study was to conjugate and evaluate methotrexate with C60-fullerenes and multi-walled carbon nanotubes (MWCNTs) for better drug delivery to cancer cells, and also to compare these two systems. C60-fullerenes and MWCNTs were functionalized by 1,3-dipolar cycloaddition using glycine and paraformaldehyde. Methotrexate (MTX) was esterified and conjugated to the functionalized carbon-based carriers. The conjugates were characterized for micromeritics and drug conjugation. The systems were evaluated for drug release in various pH, MTT cytotoxicity assay, protein binding, cellular uptake, haemolytic profile and pharmacokinetics. Spectroscopic studies confirmed the successful conjugation of drug to the aminated carbon-based carriers. The developed systems released more drug at the pH of cancer cells to that of the pH of plasma. The carriers were compatible with erythrocytes and offered substantial cytotoxicity. Better cellular uptake was confirmed by confocal laser scanning microscopy. C60-fullerenes/MWCNTs modulated the pharmacokinetic profile of drug in desired manner, resulting in better retention and compartment availability. However, the results from C60-fullerenes were observed to be better than that from MWCNTs. The present findings established the potential of carbon-based aminated nanocarriers for delivery of methotrexate in safer and effective manner. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Encapsulation of Polymethoxyflavones in Citrus Oil Emulsion-Based Delivery Systems.

    Science.gov (United States)

    Yang, Ying; Zhao, Chengying; Chen, Jingjing; Tian, Guifang; McClements, David Julian; Xiao, Hang; Zheng, Jinkai

    2017-03-01

    The purpose of this work was to elucidate the effects of citrus oil type on polymethoxyflavone (PMF) solubility and on the physicochemical properties of PMF-loaded emulsion-based delivery systems. Citrus oils were extracted from mandarin, orange, sweet orange, and bergamot. The major constituents were determined by GC/MS: sweet orange oil (97.4% d-limonene); mandarin oil (72.4% d-limonene); orange oil (67.2% d-limonene); and bergamot oil (34.6% linalyl acetate and 25.3% d-limonene). PMF-loaded emulsions were fabricated using 10% oil phase (containing 0.1% w/v nobiletin or tangeretin) and 90% aqueous phase (containing 1% w/v Tween 80) using high-pressure homogenization. Delivery systems prepared using mandarin oil had the largest mean droplet diameters (386 or 400 nm), followed by orange oil (338 or 390 nm), bergamot oil (129 or 133 nm), and sweet orange oil (122 or 126 nm) for nobiletin- or tangeretin-loaded emulsions, respectively. The optical clarity of the emulsions increased with decreasing droplet size due to reduced light scattering. The viscosities of the emulsions (with or without PMFs) were similar (1.3 to 1.4 mPa·s), despite appreciable differences in oil phase viscosity. The loading capacity and encapsulation efficiency of the emulsions depended on carrier oil type, with bergamot oil giving the highest loading capacity. In summary, differences in the composition and physical characteristics of citrus oils led to PMF-loaded emulsions with different encapsulation and physicochemical characteristics. These results will facilitate the rational design of emulsion-based delivery systems for encapsulation of PMFs and other nutraceuticals in functional foods and beverages.

  17. Development of a Multivalent Subunit Vaccine against Tularemia Using Tobacco Mosaic Virus (TMV Based Delivery System.

    Directory of Open Access Journals (Sweden)

    Sukalyani Banik

    Full Text Available Francisella tularensis is a facultative intracellular pathogen, and is the causative agent of a fatal human disease known as tularemia. F. tularensis is classified as a Category A Biothreat agent by the CDC based on its use in bioweapon programs by several countries in the past and its potential to be used as an agent of bioterrorism. No licensed vaccine is currently available for prevention of tularemia. In this study, we used a novel approach for development of a multivalent subunit vaccine against tularemia by using an efficient tobacco mosaic virus (TMV based delivery platform. The multivalent subunit vaccine was formulated to contain a combination of F. tularensis protective antigens: OmpA-like protein (OmpA, chaperone protein DnaK and lipoprotein Tul4 from the highly virulent F. tularensis SchuS4 strain. Two different vaccine formulations and immunization schedules were used. The immunized mice were challenged with lethal (10xLD100 doses of F. tularensis LVS on day 28 of the primary immunization and observed daily for morbidity and mortality. Results from this study demonstrate that TMV can be used as a carrier for effective delivery of multiple F. tularensis antigens. TMV-conjugate vaccine formulations are safe and multiple doses can be administered without causing any adverse reactions in immunized mice. Immunization with TMV-conjugated F. tularensis proteins induced a strong humoral immune response and protected mice against respiratory challenges with very high doses of F. tularensis LVS. This study provides a proof-of-concept that TMV can serve as a suitable platform for simultaneous delivery of multiple protective antigens of F. tularensis. Refinement of vaccine formulations coupled with TMV-targeting strategies developed in this study will provide a platform for development of an effective tularemia subunit vaccine as well as a vaccination approach that may broadly be applicable to many other bacterial pathogens.

  18. IgG-loaded hyaluronan-based dissolving microneedles for intradermal protein delivery.

    Science.gov (United States)

    Mönkäre, Juha; Reza Nejadnik, M; Baccouche, Khalil; Romeijn, Stefan; Jiskoot, Wim; Bouwstra, Joke A

    2015-11-28

    Dissolving microneedles are an attractive approach for non-invasive delivery of drugs via the skin, particularly when the doses are in the microgram or low-milligram range. The aim of the study was to develop hyaluronan-based, monoclonal IgG-loaded microneedles for intradermal delivery enabling efficient penetration and rapid dissolution in the skin while preserving protein stability. Microscopic analysis showed successful preparation of sharp microneedles with the tip length of ~280 μm and with up to 10% (w/w) of IgG content. The water content of the microneedles was ~12% and was not affected by the protein content. The protein distribution was uniform within microneedle tips and individual arrays but some array-to-array variation of IgG level within a single preparation batch was detected. After dissolution of microneedle arrays in PBS, N80% of protein was recovered and no conformational changes were detected by fluorescence spectroscopy. At submicron level, only weak and reversible interaction between HA and IgG was found by asymmetric flow field flow fractionation analysis after the dissolution of prepared microneedles. Although, the formation of insoluble micron-size particles was detected by flow imaging microscopy the IgG amount incorporated into these particles was negligible (b5%). Finally, microneedles were able to penetrate into the epidermis of ex vivo human skin followed by the rapid dissolution of the microneedle tips in the skin. After 10 min of application, the majority of the original tip length was dissolved and IgG and hyaluronan were co-deposited until a depth of 150-200 μm in the skin. In conclusion, developed hyaluronan-based dissolving microneedles allow rapid noninvasive intradermal protein delivery.

  19. Fibrin matrices with affinity-based delivery systems and neurotrophic factors promote functional nerve regeneration.

    Science.gov (United States)

    Wood, Matthew D; MacEwan, Matthew R; French, Alexander R; Moore, Amy M; Hunter, Daniel A; Mackinnon, Susan E; Moran, Daniel W; Borschel, Gregory H; Sakiyama-Elbert, Shelly E

    2010-08-15

    Glial-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) have both been shown to enhance peripheral nerve regeneration following injury and target different neuronal populations. The delivery of either growth factor at the site of injury may, therefore, result in quantitative differences in motor nerve regeneration and functional recovery. In this study we evaluated the effect of affinity-based delivery of GDNF or NGF from fibrin-filled nerve guidance conduits (NGCs) on motor nerve regeneration and functional recovery in a 13 mm rat sciatic nerve defect. Seven experimental groups were evaluated consisting of GDNF or NGF and the affinity-based delivery system (DS) within NGCs, control groups excluding the DS and/or growth factor, and nerve isografts. Groups with growth factor in the conduit demonstrated equivalent or superior performance in behavioral tests and relative muscle mass measurements compared to isografts at 12 weeks. Additionally, groups with GDNF demonstrated greater specific twitch and tetanic force production in extensor digitorum longus (EDL) muscle than the isograft control, while groups with NGF produced demonstrated similar force production compared to the isograft control. Assessment of motor axon regeneration by retrograde labeling further revealed that the number of ventral horn neurons regenerating across NGCs containing GDNF and NGF DS was similar to the isograft group and these counts were greater than the groups without growth factor. Overall, the GDNF DS group demonstrated superior functional recovery and equivalent motor nerve regeneration compared to the isograft control, suggesting it has potential as a treatment for motor nerve injury.

  20. Bystander Effect Induced by Electroporation is Possibly Mediated by Microvesicles and Dependent on Pulse Amplitude, Repetition Frequency and Cell Type.

    Science.gov (United States)

    Prevc, Ajda; Bedina Zavec, Apolonija; Cemazar, Maja; Kloboves-Prevodnik, Veronika; Stimac, Monika; Todorovic, Vesna; Strojan, Primoz; Sersa, Gregor

    2016-10-01

    Bystander effect, a known phenomenon in radiation biology, where irradiated cells release signals which cause damage to nearby, unirradiated cells, has not been explored in electroporated cells yet. Therefore, our aim was to determine whether bystander effect is present in electroporated melanoma cells in vitro, by determining viability of non-electroporated cells exposed to medium from electroporated cells and by the release of microvesicles as potential indicators of the bystander effect. Here, we demonstrated that electroporation of cells induces bystander effect: Cells exposed to electric pulses mediated their damage to the non-electroporated cells, thus decreasing cell viability. We have shown that shedding microvesicles may be one of the ways used by the cells to mediate the death signals to the neighboring cells. The murine melanoma B16F1 cell line was found to be more electrosensitive and thus more prone to bystander effect than the canine melanoma CMeC-1 cell line. In B16F1 cell line, bystander effect was present above the level of electropermeabilization of the cells, with the threshold at 800 V/cm. Furthermore, with increasing electric field intensities and the number of pulses, the bystander effect also increased. In conclusion, electroporation can induce bystander effect which may be mediated by microvesicles, and depends on pulse amplitude, repetition frequency and cell type.

  1. Injection molded chips with integrated conducting polymer electrodes for electroporation of cells

    DEFF Research Database (Denmark)

    Andresen, Kristian; Hansen, Morten; Matschuk, Maria

    2010-01-01

    We present the design-concept for an all polymer injection molded single use microfluidic device. The fabricated devices comprise integrated conducting polymer electrodes and Luer fitting ports to allow for liquid and electrical access. A case study of low voltage electroporation of biological...

  2. High-Voltage Electroporation of Bacteria: Genetic Transformation of Campylobacter jejuni with Plasmid DNA

    Science.gov (United States)

    Miller, Jeff F.; Dower, William J.; Tompkins, Lucy S.

    1988-02-01

    Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of high-voltage discharges to bacterial cells permits genetic transformation. Our method involves exposure of a Campylobacter cell suspension to a high-voltage exponential decay discharge (5-13 kV/cm) for a brief period of time (resistance-capacitance time constant = 2.4-26 msec) in the presence of plasmid DNA. Electrical transformation of C. jejuni results in frequencies as high as 1.2 × 106 transformants per μ g of DNA. We have investigated the effects of pulse amplitude and duration, cell growth conditions, divalent cations, and DNA concentration on the efficiency of transformation. Transformants of C. jejuni obtained by electroporation contained structurally intact plasmid molecules. In addition, evidence is presented that indicates that C. jejuni possesses DNA restriction and modification systems. The use of electroporation as a method for transforming other bacterial species and guidelines for its implementation are also discussed.

  3. Irreversible electroporation ablation area enhanced by synergistic high- and low-voltage pulses.

    Directory of Open Access Journals (Sweden)

    Chenguo Yao

    Full Text Available Irreversible electroporation (IRE produced by a pulsed electric field can ablate tissue. In this study, we achieved an enhancement in ablation area by using a combination of short high-voltage pulses (HVPs to create a large electroporated area and long low-voltage pulses (LVPs to ablate the electroporated area. The experiments were conducted in potato tuber slices. Slices were ablated with an array of four pairs of parallel steel electrodes using one of the following four electric pulse protocols: HVP, LVP, synergistic HVP+LVP (SHLVP or LVP+HVP. Our results showed that the SHLVPs more effectively necrotized tissue than either the HVPs or LVPs, even when the SHLVP dose was the same as or lower than the HVP or LVP doses. The HVP and LVP order mattered and only HVPs+LVPs (SHLVPs treatments increased the size of the ablation zone because the HVPs created a large electroporated area that was more susceptible to the subsequent LVPs. Real-time temperature change monitoring confirmed that the tissue was non-thermally ablated by the electric pulses. Theoretical calculations of the synergistic effects of the SHLVPs on tissue ablation were performed. Our proposed SHLVP protocol provides options for tissue ablation and may be applied to optimize the current clinical IRE protocols.

  4. Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform.

    Science.gov (United States)

    Madison, Andrew C; Royal, Matthew W; Vigneault, Frederic; Chen, Liji; Griffin, Peter B; Horowitz, Mark; Church, George M; Fair, Richard B

    2017-09-15

    Electrowetting-on-dielectric (EWD) digital microfluidic laboratory-on-a-chip platforms demonstrate excellent performance in automating labor-intensive protocols. When coupled with an on-chip electroporation capability, these systems hold promise for streamlining cumbersome processes such as multiplex automated genome engineering (MAGE). We integrated a single Ti:Au electroporation electrode into an otherwise standard parallel-plate EWD geometry to enable high-efficiency transformation of Escherichia coli with reporter plasmid DNA in a 200 nL droplet. Test devices exhibited robust operation with more than 10 transformation experiments performed per device without cross-contamination or failure. Despite intrinsic electric-field nonuniformity present in the EP/EWD device, the peak on-chip transformation efficiency was measured to be 8.6 ± 1.0 × 108 cfu·μg-1 for an average applied electric field strength of 2.25 ± 0.50 kV·mm-1. Cell survival and transformation fractions at this electroporation pulse strength were found to be 1.5 ± 0.3 and 2.3 ± 0.1%, respectively. Our work expands the EWD toolkit to include on-chip microbial electroporation and opens the possibility of scaling advanced genome engineering methods, like MAGE, into the submicroliter regime.

  5. Irreversible electroporation ablation area enhanced by synergistic high- and low-voltage pulses.

    Science.gov (United States)

    Yao, Chenguo; Lv, Yanpeng; Dong, Shoulong; Zhao, Yajun; Liu, Hongmei

    2017-01-01

    Irreversible electroporation (IRE) produced by a pulsed electric field can ablate tissue. In this study, we achieved an enhancement in ablation area by using a combination of short high-voltage pulses (HVPs) to create a large electroporated area and long low-voltage pulses (LVPs) to ablate the electroporated area. The experiments were conducted in potato tuber slices. Slices were ablated with an array of four pairs of parallel steel electrodes using one of the following four electric pulse protocols: HVP, LVP, synergistic HVP+LVP (SHLVP) or LVP+HVP. Our results showed that the SHLVPs more effectively necrotized tissue than either the HVPs or LVPs, even when the SHLVP dose was the same as or lower than the HVP or LVP doses. The HVP and LVP order mattered and only HVPs+LVPs (SHLVPs) treatments increased the size of the ablation zone because the HVPs created a large electroporated area that was more susceptible to the subsequent LVPs. Real-time temperature change monitoring confirmed that the tissue was non-thermally ablated by the electric pulses. Theoretical calculations of the synergistic effects of the SHLVPs on tissue ablation were performed. Our proposed SHLVP protocol provides options for tissue ablation and may be applied to optimize the current clinical IRE protocols.

  6. Thermal Energy during Irreversible Electroporation and the Influence of Different Ablation Parameters

    NARCIS (Netherlands)

    van den Bos, Willemien; Scheffer, Hester J.; Vogel, Jantien A.; Wagstaff, Peter G. K.; de Bruin, Daniel M.; de Jong, Marcus C.; van Gemert, Martin J. C.; de La Rosette, Jean J. M. C. H.; Meijerink, Martijn R.; Klaessens, John H.; Verdaasdonk, Rudolf M.

    2016-01-01

    Irreversible electroporation (IRE) uses high-voltage electric fields to achieve cell death. Although the mechanism of IRE is mainly designated as nonthermal, development of secondary Joule heating is inevitable. The study purpose was to gain understanding of temperature development and distribution

  7. Electroporation of Alcaligenes eutrophus with (mega) plasmids and genomic DNA fragments.

    Science.gov (United States)

    Taghavi, S; van der Lelie, D; Mergeay, M

    1994-10-01

    Electroporation was used as a tool to explore the genetics of the heavy-metal-resistant strain Alcaligenes eutrophus CH34. A 12.9-kb A. eutrophus-Escherichia coli shuttle vector, pMOL850, was constructed to optimize electroporation conditions. This vector is derived from the E. coli plasmid pSUP202 and contains the replication region of the A. eutrophus megaplasmid pMOL28. Electroporation was used to transform A. eutrophus CH34 derivatives with megaplasmids (sizes up to 240 kb), and transformants were selected for resistance to heavy metals. Electroporation was also performed with endonuclease-digested genomic DNA. Transformation of markers affecting lysine biosynthesis (lysA194) and biosynthesis of the siderophore alcaligin E were observed. Transfer of the nonselected markers pheB332 and aro-333, linked to lysA194, confirmed the intervention of homologous recombination. However, during transformation of ale::Tn5-Tc, illegitimate recombination and transposition were also observed as an alternative for the inheritance of the Tn5-Tc markers.

  8. Loading of acute myeloid leukemia cells with poly(I:C) by electroporation

    NARCIS (Netherlands)

    Lion, E.; Winde, C.M. de; Tendeloo, V.F. Van; Smits, E.L.

    2014-01-01

    In this chapter, we describe the technique of electroporation as an efficient method to load primary leukemic cells with the double-stranded RNA (dsRNA) analogue, polyriboinosinic polyribocytidylic acid (poly(I:C)), and detail on the delicate freezing and thawing procedure of primary leukemic

  9. Calcium Electroporation: Evidence for Differential Effects in Normal and Malignant Cell Lines, Evaluated in a 3D Spheroid Model.

    Science.gov (United States)

    Frandsen, Stine Krog; Gibot, Laure; Madi, Moinecha; Gehl, Julie; Rols, Marie-Pierre

    2015-01-01

    Calcium electroporation describes the use of high voltage electric pulses to introduce supraphysiological calcium concentrations into cells. This promising method is currently in clinical trial as an anti-cancer treatment. One very important issue is the relation between tumor cell kill efficacy-and normal cell sensitivity. Using a 3D spheroid cell culture model we have tested the effect of calcium electroporation and electrochemotherapy using bleomycin on three different human cancer cell lines: a colorectal adenocarcinoma (HT29), a bladder transitional cell carcinoma (SW780), and a breast adenocarcinoma (MDA-MB231), as well as on primary normal human dermal fibroblasts (HDF-n). The results showed a clear reduction in spheroid size in all three cancer cell spheroids three days after treatment with respectively calcium electroporation (pspheroids was neither affected after calcium electroporation nor electrochemotherapy using bleomycin, indicating that calcium electroporation, like electrochemotherapy, will have limited adverse effects on the surrounding normal tissue when treating with calcium electroporation. The intracellular ATP level, which has previously been shown to be depleted after calcium electroporation, was measured in the spheroids after treatment. The results showed a dramatic decrease in the intracellular ATP level (pspheroid types-malignant as well as normal. In conclusion, calcium electroporation seems to be more effective in inducing cell death in cancer cell spheroids than in a normal fibroblast spheroid, even though intracellular ATP level is depleted in all spheroid types after treatment. These results may indicate an important therapeutic window for this therapy; although further studies are needed in vivo and in patients to investigate the effect of calcium electroporation on surrounding normal tissue when treating tumors.

  10. Poly(lactic-co-glycolic) Acid/Solutol HS15-Based Nanoparticles for Docetaxel Delivery.

    Science.gov (United States)

    Cho, Hyun-Jong; Park, Ju-Hwan; Kim, Dae-Duk; Yoon, In-Soo

    2016-02-01

    Docetaxel (DCT) is one of anti-mitotic chemotherapeutic agents and has been used for the treatment of gastric cancer as well as head and neck cancer, breast cancer and prostate cancer. Poly(lactic- co-glycolic) acid (PLGA) is one of representative biocompatible and biodegradable polymers, and polyoxyl 15 hydroxystearate (Solutol HS15) is a nonionic solubilizer and emulsifying agent. In this investigation, PLGA/Solutol HS15-based nanoparticles (NPs) for DCT delivery were fabricated by a modified emulsification-solvent evaporation method. PLGA/Solutol HS15/DCT NPs with about 169 nm of mean diameter, narrow size distribution, negative zeta potential, and spherical morphology were prepared. The results of solid-state studies revealed the successful dispersion of DCT in PLGA matrix and its amorphization during the preparation process of NPs. According to the result of in vitro release test, emulsifying property of Solutol HS15 seemed to contribute to the enhanced drug release from NPs at physiological pH. All these findings imply that developed PLGA/Solutol HS15-based NP can be a promising local anticancer drug delivery system for cancer therapy.

  11. Fabrication and characterization of sol-gel based nanoparticles for drug delivery

    Science.gov (United States)

    Yadav, Reeta

    Nanogels are cross linked polymeric sol-gel based nanoparticles that offer an interior network for incorporation and protection of biomolecules, exhibiting unique advantages for polymer based delivery systems. We have successfully synthesized stable sol-gel nanoparticles by means of [a] silicification reactions using cationic peptides like polylysine as gelating agents, and [b] lyophilization of sol-gels. Macromolecules such as Hemoglobin and Glucose Oxidase and small molecules such as Sodium Nitroprusside (SNP) and antibiotics were encapsulated within the nanogels. We have used transmission electron microscopy, dynamic light scattering, zeta potential analysis, and spectroscopy to perform a physicochemical characterization of the nanogels resulting from the two approaches. Our studies have indicated that the nanogel encapsulated proteins and small molecules remain intact, stable and functional. A Hydrogen Peroxide (H2O2) and Nitric Oxide (NO) generating drug carrier was synthesized using these nanogels and the effect of generation of H2O2 from Glucose Oxidase encapsulated nanogels and NO from SNP encapsulated nanogels was tested on E.coli. The results show that the nanoparticles exert antimicrobial activity against E.Coli, in addition NO generating nanogels potentiated H2O2 generating nanogels induced killing. These data suggest that these NO and H2O2 releasing nanogels have the potential to serve as a novel class of antimicrobials for the treatment of multidrug resistant bacteria. The unique properties of these protein/drug incorporated nanogels raise the prospect of fine tailoring to specific applications such as drug delivery and bio imaging.

  12. Operationalising emergency care delivery in sub-Saharan Africa: consensus-based recommendations for healthcare facilities.

    Science.gov (United States)

    Calvello, Emilie J B; Tenner, Andrea G; Broccoli, Morgan C; Skog, Alexander P; Muck, Andrew E; Tupesis, Janis P; Brysiewicz, Petra; Teklu, Sisay; Wallis, Lee; Reynolds, Teri

    2016-08-01

    A major barrier to successful integration of acute care into health systems is the lack of consensus on the essential components of emergency care within resource-limited environments. The 2013 African Federation of Emergency Medicine Consensus Conference was convened to address the growing need for practical solutions to further implementation of emergency care in sub-Saharan Africa. Over 40 participants from 15 countries participated in the working group that focused on emergency care delivery at health facilities. Using the well-established approach developed in the WHO's Monitoring Emergency Obstetric Care, the workgroup identified the essential services delivered-signal functions-associated with each emergency care sentinel condition. Levels of emergency care were assigned based on the expected capacity of the facility to perform signal functions, and the necessary human, equipment and infrastructure resources identified. These consensus-based recommendations provide the foundation for objective facility capacity assessment in developing emergency health systems that can bolster strategic planning as well as facilitate monitoring and evaluation of service delivery. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  13. A novel subcutaneous infusion delivery system based on osmotic pump: in vitro and in vivo evaluation.

    Science.gov (United States)

    Gong, Wei; Ma, Rui; Mei, Danyu; Jing, Pei; Dong, Xiao; Li, Bingsheng; Yang, Yanfang; Du, Lina; Mei, Xing-Guo; Hu, Fu-Qiang

    2014-02-01

    An economical, convenient portable drug delivery system combining osmotic pump with subcutaneous infusion was developed, which was composed of three primary components: water chamber, osmotic pump chamber and support base. Ceftriaxone sodium (CRO) was selected as the model drug and osmotic pump tablets were prepared. The influence of osmotic agents on drug release profiles was evaluated. As the adjustment made by the osmotic agents was limited, the compositions of semipermeable membrane were investigated to determine significant associations of factors based on orthogonal design. The in vitro release profiles of the optimum formulation achieved to the predetermined value (15 ± 3 min for the initial release time T(i) and 5.75 ± 0.25 h for the extent release time T(e)). The pharmacokinetic profiles of this drug delivery system were evaluated in Beagle dogs. In vivo results demonstrated that the osmotic pump subcutaneous infusion administration was equivalent to intravenous injection administration in terms of bioavailability. Moreover, constant drug plasma levels with minimized fluctuations could be achieved with this osmotic pump subcutaneous infusion system, compared with intravenous injection.

  14. Nanoparticle-based drug delivery systems: a commercial and regulatory outlook as the field matures.

    Science.gov (United States)

    Ragelle, Héloïse; Danhier, Fabienne; Préat, Véronique; Langer, Robert; Anderson, Daniel G

    2017-07-01

    Nanomedicine has emerged as a major field of academic research with direct impact on human health. While a first generation of products has been successfully commercialized and has significantly contributed to enhance patient's life, recent advances in material design and the emergence of new therapeutics are contributing to the development of more sophisticated systems. As the field matures, it is important to comprehend the challenges related to nanoparticle commercial development for a more efficient and predictable path to the clinic. Areas covered: The review provides an overview of nanoparticle-based delivery systems currently on the market and in clinical trials, and discuss the principal challenges for their commercial development, both from a manufacturing and regulatory perspective, to help gain understanding of the translational path for these systems. Expert opinion: Clinical translation of nanoparticle-based delivery systems remains challenging on account of their 3D nanostructure and requires robust nano-manufacturing process along with adequate analytical tools and methodologies. By identifying early enough in the development the product critical attributes and understanding their impact on the therapeutic performance, the developers of nanopharmaceuticals will be better equipped to develop efficient product pipelines. Second-generation products are expected to broaden nanopharmaceutical global market in the upcoming years.

  15. Heterosubtypic protection against pathogenic human and avian influenza viruses via in vivo electroporation of synthetic consensus DNA antigens.

    Directory of Open Access Journals (Sweden)

    Dominick J Laddy

    Full Text Available BACKGROUND: The persistent evolution of highly pathogenic avian influenza (HPAI highlights the need for novel vaccination techniques that can quickly and effectively respond to emerging viral threats. We evaluated the use of optimized consensus influenza antigens to provide broad protection against divergent strains of H5N1 influenza in three animal models of mice, ferrets, and non-human primates. We also evaluated the use of in vivo electroporation to deliver these vaccines to overcome the immunogenicity barrier encountered in larger animal models of vaccination. METHODS AND FINDINGS: Mice, ferrets and non-human primates were immunized with consensus plasmids expressing H5 hemagglutinin (pH5HA, N1 neuraminidase (pN1NA, and nucleoprotein antigen (pNP. Dramatic IFN-gamma-based cellular immune responses to both H5 and NP, largely dependent upon CD8+ T cells were seen in mice. Hemaggutination inhibition titers classically associated with protection (>1:40 were seen in all species. Responses in both ferrets and macaques demonstrate the ability of synthetic consensus antigens to induce antibodies capable of inhibiting divergent strains of the H5N1 subtype, and studies in the mouse and ferret demonstrate the ability of synthetic consensus vaccines to induce protection even in the absence of such neutralizing antibodies. After challenge, protection from morbidity and mortality was seen in mice and ferrets, with significant reductions in viral shedding and disease progression seen in vaccinated animals. CONCLUSIONS: By combining several consensus influenza antigens with in vivo electroporation, we demonstrate that these antigens induce both protective cellular and humoral immune responses in mice, ferrets and non-human primates. We also demonstrate the ability of these antigens to protect from both morbidity and mortality in a ferret model of HPAI, in both the presence and absence of neutralizing antibody, which will be critical in responding to the

  16. Surface bioengineering of diatomite based nanovectors for efficient intracellular uptake and drug delivery

    Science.gov (United States)

    Terracciano, Monica; Shahbazi, Mohammad-Ali; Correia, Alexandra; Rea, Ilaria; Lamberti, Annalisa; de Stefano, Luca; Santos, Hélder A.

    2015-11-01

    Diatomite is a natural porous silica material of sedimentary origin. Due to its peculiar properties, it can be considered as a valid surrogate of synthetic porous silica for nano-based drug delivery. In this work, we exploit the potential of diatomite nanoparticles (DNPs) for drug delivery with the aim of developing a successful dual-biofunctionalization method by polyethylene glycol (PEG) coverage and cell-penetrating peptide (CPP) bioconjugation, to improve the physicochemical and biological properties of the particles, to enhance the intracellular uptake in cancer cells, and to increase the biocompatibility of 3-aminopropyltriethoxysilane (APT) modified-DNPs. DNPs-APT-PEG-CPP showed hemocompatibility for up to 200 μg mL-1 after 48 h of incubation with erythrocytes, with a hemolysis value of only 1.3%. The cytotoxicity of the modified-DNPs with a concentration up to 200 μg mL-1 and incubation with MCF-7 and MDA-MB-231 breast cancer cells for 24 h, demonstrated that PEGylation and CPP-bioconjugation can strongly reduce the cytotoxicity of DNPs-APT. The cellular uptake of the modified-DNPs was also evaluated using the above mentioned cancer cell lines, showing that the CPP-bioconjugation can considerably increase the DNP cellular uptake. Moreover, the dual surface modification of DNPs improved both the loading of a poorly water-soluble anticancer drug, sorafenib, with a loading degree up to 22 wt%, and also enhanced the drug release profiles in aqueous solutions. Overall, this work demonstrates that the biofunctionalization of DNPs is a promising platform for drug delivery applications in cancer therapy as a result of its enhanced stability, biocompatibility, cellular uptake, and drug release profiles.Diatomite is a natural porous silica material of sedimentary origin. Due to its peculiar properties, it can be considered as a valid surrogate of synthetic porous silica for nano-based drug delivery. In this work, we exploit the potential of diatomite nanoparticles

  17. Two cholesterol derivative-based PEGylated liposomes as drug delivery system, study on pharmacokinetics and drug delivery to retina

    Science.gov (United States)

    Geng, Shengyong; Yang, Bin; Wang, Guowu; Qin, Geng; Wada, Satoshi; Wang, Jin-Ye

    2014-07-01

    In this study, two cholesterol derivatives, (4-cholesterocarbonyl-4‧-(N,N,N-triethylamine butyloxyl bromide) azobenzene (CAB) and 4-cholesterocarbonyl-4‧-(N,N-diethylamine butyloxyl) azobenzene (ACB), one of which is positively charged while the other is neutral, were synthesized and incorporated with phospholipids and cholesterol to form doxorubicin (DOX)-loaded liposomes. PEGylation was achieved by including 1,2-distearoyl-sn-glycero-3-phosphatiylethanol-amine-N-[methoxy-(polyethylene glycol)-2000 (DSPE-PEG2000). Our results showed that PEGylated liposomes displayed significantly improved stability and the drug leakage was decreased compared to the non-PEGylated ones in vitro. The in vivo study with rats also revealed that the pharmacokinetics and circulation half-life of DOX were significantly improved when liposomes were PEGylated (p derivative ACB played some role in improving liposomes’ stability in systemic circulation compared to the conventional PC liposome and the positively charged CAB liposome, with or without PEGylation. In addition, in the case of local drug delivery, the positively charged PEG-liposome not only delivered much more of the drug into the rats’ retinas (p < 0.001), but also maintained much longer drug retention time compared to the neutral PEGylated liposomes.

  18. Light-sensitive Lipid-based Nanoparticles for Drug Delivery: Design Principles and Future Considerations for Biological Applications

    OpenAIRE

    Yavlovich, Amichai; Smith, Brandon; Gupta, Kshitij; Blumenthal, Robert; Puri, Anu

    2010-01-01

    Radiation-based therapies aided by nanoparticles have been developed since decades, and can be primarily categorized into two main platforms. First, delivery of payload of photo-reactive drugs (photosensitizers) using the conventional nanoparticles, and second, design and development of photo-triggerable nanoparticles (primarily liposomes) to attain light-assisted on-demand drug delivery. The main focus of this review is to provide an update of the history, current status and future applicati...

  19. Efficient large volume electroporation of dendritic cells through micrometer scale manipulation of flow in a disposable polymer chip

    DEFF Research Database (Denmark)

    Selmeczi, David; Hansen, Thomas; Met, Özcan

    2011-01-01

    We present a hybrid chip of polymer and stainless steel designed for high-throughput continuous electroporation of cells in suspension. The chip is constructed with two parallel stainless steel mesh electrodes oriented perpendicular to the liquid flow. The relatively high hydrodynamic resistance...... of the chip is similar to that of the traditional electroporation cuvette, but without an upper limit on the number of cells to be electroporated. The device is constructed with two female Luer parts and can easily be integrated with other microfluidic components. Furthermore it is fabricated from injection...

  20. Transient gene expression in electroporated banana (Musa spp., cv. 'Bluggoe', ABB group) protoplasts isolated from regenerable embryogenetic cell suspensions.

    Science.gov (United States)

    Sagi, L; Remy, S; Panis, B; Swennen, R; Volckaert, G

    1994-02-01

    Electroporation conditions were established for transient expression of introduced DNA in banana (Musa spp., cv. 'Bluggoe') protoplasts isolated from regenerable embryogenic cell suspensions. The following parameters were found to be highly influential: electroporation buffer, polyethylene glycol treatment and its duration before electroporation, use of a heat shock, and chimaeric gene constructs. The maximum frequency of DNA introduction as detected by an in situ assay for transient expression of the uidA gene, amounted to 1.8% of total protoplasts. Since plants have recently been regenerated from banana protoplasts at a high frequency, the present results may contribute to the production of transgenic banana.

  1. Development of a single cell electroporation method using a scanning ion conductance microscope with a theta nanopipette

    Science.gov (United States)

    Sakurai, Satoshi; Yamazaki, Koji; Ushiki, Tatsuo; Iwata, Futoshi

    2015-08-01

    We developed a novel electroporation method using a scanning ion conductance microscope (SICM) with a theta capillary nanopipette probe that has two apertures at the edge of the pipette. One aperture of the pipette probe was used to control the pipette-surface distance and to apply pulse voltage for electroporation. The other was used to eject material over the cell by local electrophoresis. Using the nanopipette, propidium iodide was successfully introduced into a targeted single Hela cell without influencing the surrounding cells. Furthermore, by scanning the theta nanopipette probe using the SICM, the morphological behaviors of the electroporated cells could be observed.

  2. Increased gene delivery efficiency and specificity of a lipid-based nanosystem incorporating a glycolipid.

    Science.gov (United States)

    Magalhães, Mariana; Farinha, Dina; Pedroso de Lima, Maria Conceição; Faneca, Henrique

    2014-01-01

    Hepatocellular carcinoma (HCC) is the third most common cause of death related to cancer diseases worldwide. The current treatment options have many limitations and reduced success rates. In this regard, advances in gene therapy have shown promising results in novel therapeutic strategies. However, the success of gene therapy depends on the efficient and specific delivery of genetic material into target cells. In this regard, the main goal of this work was to develop a new lipid-based nanosystem formulation containing the lipid lactosyl-PE for specific and efficient gene delivery into HCC cells. The obtained results showed that incorporation of 15% of lactosyl-PE into liposomes induces a strong potentiation of lipoplex biological activity in HepG2 cells, not only in terms of transgene expression levels but also in terms of percentage of transfected cells. In the presence of galactose, which competes with lactosyl-PE for the binding to the asialoglycoprotein receptor (ASGP-R), a significant reduction in biological activity was observed, showing that the potentiation of transfection induced by the presence of lactosyl-PE could be due to its specific interaction with ASGP-R, which is overexpressed in HCC. In addition, it was found that the incorporation of lactosyl-PE in the nanosystems promotes an increase in their cell binding and uptake. Regarding the physicochemical properties of lipoplexes, the presence of lactosyl-PE resulted in a significant increase in DNA protection and in a substantial decrease in their mean diameter and zeta potential, conferring them suitable characteristics for in vivo application. Overall, the results obtained in this study suggest that the potentiation of the biological activity induced by the presence of lactosyl-PE is due to its specific binding to the ASGP-R, showing that this novel formulation could constitute a new gene delivery nanosystem for application in therapeutic strategies in HCC.

  3. Cyclen-based cationic lipids for highly efficient gene delivery towards tumor cells.

    Directory of Open Access Journals (Sweden)

    Qing-Dong Huang

    Full Text Available Gene therapy has tremendous potential for both inherited and acquired diseases. However, delivery problems limited their clinical application, and new gene delivery vehicles with low cytotoxicity and high transfection efficiency are greatly required.In this report, we designed and synthesized three amphiphilic molecules (L1-L3 with the structures involving 1, 4, 7, 10-tetraazacyclododecane (cyclen, imidazolium and a hydrophobic dodecyl chain. Their interactions with plasmid DNA were studied via electrophoretic gel retardation assays, fluorescent quenching experiments, dynamic light scattering and transmission electron microscopy. The in vitro gene transfection assay and cytotoxicity assay were conducted in four cell lines.Results indicated that L1 and L3-formed liposomes could effectively bind to DNA to form well-shaped nanoparticles. Combining with neutral lipid DOPE, L3 was found with high efficiency in gene transfer in three tumor cell lines including A549, HepG2 and H460. The optimized gene transfection efficacy of L3 was nearly 5.5 times more efficient than that of the popular commercially available gene delivery agent Lipofectamine 2000™ in human lung carcinoma cells A549. In addition, since L1 and L3 had nearly no gene transfection performance in normal cells HEK293, these cationic lipids showed tumor cell-targeting property to a certain extent. No significant cytotoxicity was found for the lipoplexes formed by L1-L3, and their cytotoxicities were similar to or slightly lower than the lipoplexes prepared from Lipofectamine 2000™.Novel cyclen-based cationic lipids for effective in vitro gene transfection were founded, and these studies here may extend the application areas of macrocyclic polyamines, especially for cyclen.

  4. GIS-based Mine Tailings Yield Mapping using RUSLE and Sediment Delivery Ratio

    Science.gov (United States)

    Kim, S.; Choi, Y.; Park, H.; Kwon, H.; Yoon, S.; Go, W.

    2010-12-01

    Erosion of mine tailings heaped up on the side of abandoned mine is an environmental problem because they contain harmful heavy metals. These harmful heavy metals such as copper, lead, arsenic in mine tailings cause contamination of surrounding streams and soil. To prevent and reduce the damage of surrounding streams caused by harmful heavy metals leaking from mine tailings, evaluating the pollution loading amount of mine tailings is required. However, it is difficult to assess its environmental impacts accurately because of its complex processes associated with it (Lal 1994). To estimate soil erosion and develop soil erosion management plans, there are some soil erosion estimation methods. Among these methods, the Revised Universal Soil Loss Equation (RUSLE) is the most widely used method. The six factors affecting soil loss such as rainfall-runoff erosivity, soil erodibility, slope length and steepness, cover management, and support practice were extracted from the spatial data and measurement data to evaluate average annual soil loss. Applying this model to mine tailings is possible, because mine tailings are regarded as soil. All the sediment generated may not be delivered at the watershed outlet because some of it may be deposited at various locations in the watershed. RUSLE does not consider the sediment delivery ratio to estimate the mine tailings delivered to the downstream point of interest. In this study, three methods are provided to compute the spatially distributed sediment delivery ratios and the results are compared with each other. Geographical Information System (GIS)-based erosion model and sediment delivery model were used to estimate the potential sediment yield from mine tailings in this study. The results achieved in this study can be used as basis data to assist mine tailings management and tailings dam installation plan. This work was supported by the Mine Reclamation Corporation funded by the Ministry of Knowledge Economy, Republic of Korea

  5. Strategic, value-based delivery in global health care: innovations at Harvard University and Brigham and Women's Hospital.

    Science.gov (United States)

    Weintraub, Rebecca L; Talbot, Julie; ole-MoiYoi, Kileken; Wachter, Keri; Sullivan, Erin; House, Amy; Baron, Jennifer; Beals, Aaron; Beauvais, Sophie; Rhatigan, Joseph

    2011-01-01

    Investments in global health have more than doubled over the past decade, generating a cadre of new institutions. To date, most of the funded research in global health has focused on discovery, and, more recently, on the development of new tools, which has tightened the implementation bottleneck. This article introduces the concept of global health delivery and the need to catalog and analyze current implementation efforts to bridge gaps in delivery. Global health delivery is complex and context-dependent and requires an interdisciplinary effort, including the application of strategic principles. Furthermore, delivery is necessary to ensure that the investments in research, discovery, and development generate value for patients and populations. This article discusses the application of value-based delivery to global health. It provides some examples of approaches to aggregating implicit knowledge to inform practice. With global health delivery, the aim is to transform global health scale-up from a series of well-intentioned but often disconnected efforts to a value-based movement based upon 21st-century technology, standards, and efficiency. © 2011 Mount Sinai School of Medicine.

  6. Cryopreservation of Human Adipose-Derived Stem Cells in Combination with Trehalose and Reversible Electroporation.

    Science.gov (United States)

    Dovgan, Barbara; Barlič, Ariana; Knežević, Miomir; Miklavčič, Damijan

    2017-02-01

    New cryopreservation approaches for medically applicable cells are of great importance in clinical medicine. Current protocols employ the use of dimethyl sulfoxide (DMSO), which is toxic to cells and causes undesirable side effects in patients, such as cardiac arrhythmias, neurological events, and others. Trehalose, a nontoxic disaccharide, has been already studied as a cryoprotectant. However, an efficient approach for loading this impermeable sugar into mammalian cells is missing. In our study, we assessed the efficiency of combining reversible electroporation and trehalose for cryopreservation of human adipose-derived stem cells. First, we determined reversible electroporation threshold by loading of propidium iodide into cells. The highest permeabilization while maintaining high cell viability was reached at 1.5 kV/cm, at 8 pulses, 100 µs, and 1 Hz. Second, cells were incubated in 250 or 400 mM trehalose and electroporated before cryopreservation. After thawing, 83.8 ± 1.8 % (mean ± SE) cell recovery was obtained at 250 mM trehalose. By using a standard freezing protocol (10 % DMSO in 90 % fetal bovine serum), cell survival after thawing was about 91.5 ± 1.6 %. We also evaluated possible effects of electroporation on cells' functionality before and after thawing. Successful cell growth and efficient adipogenic and osteogenic differentiation were achieved. In conclusion, electroporation seems to be an efficient method for loading nonpermeable trehalose into human adipose-derived stem cells, allowing long-term cryopreservation in DMSO-free and xeno-free conditions.

  7. Design and Characterization of Silicone and Surfactant Based Systems for Topical Drug Delivery.

    Science.gov (United States)

    Oyafuso, Márcia Helena; Carvalho, Flávia Chiva; Chiavacci, Leila Aparecida; Gremião, Maria Palmira Daflon; Chorilli, Marlus

    2015-01-01

    Nanotechnology offers advantages for new drug delivery design by providing drug targeting while minimizing the side effects. Polyoxyethylene 20 cetyl alcohol (CETETH-20) is a surfactant that may form nanostructured systems, such as liquid crystals, when in contact with water/oil, which are structurally similar to biological membranes and may improve skin interaction. The aim of this study was to develop and characterize CETETH 20-based nanostructured systems by combining CETETH-20 with water and different oily phases, including PEG-12-dimethicone for topical drug administration. The systems were characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), rheology, texture profile analyses (TPA), in vitro cytotoxicity and histopathological analyses of rabbits' skin. Lamellar, hexagonal and cubic phases were identified and their viscoelastic moduli varied according to each phase. The stiffness of the cubic phase was 3-fold higher and twice more adhesive than the hexagonal phase. The formulations did not affect the normal macrophages cells, neither promoted skin irritation. They were spontaneously obtained by simply mixing the components, which corroborates for an ease scaled-up. These results suggest that systems composed of CETETH 20, PEG-12-dimethicone and water are a promising new approach for designing nanostructured topical drug delivery systems.

  8. Development of nanotechnology-based drug delivery systems with olive vegetable oil for cutaneous application

    Directory of Open Access Journals (Sweden)

    Silas Arandas Monteiro e Silva

    Full Text Available ABSTRACT Liquid-Crystalline Systems represent active compounds delivery systems that may be able to overcome the physical barrier of the skin, especially represented by the stratum corneum. To obtain these systems, aqueous and oily components are used with surfactants. Of the different association structures in such systems, the liquid-crystalline offer numerous advantages to a topical product. This manuscript presents the development of liquid-crystalline systems consisting, in which the oil component is olive oil, its rheological characterizations, and the location of liquid crystals in its phase map. Cytotoxic effects were evaluated using J-774 mouse macrophages as the cellular model. A phase diagram to mix three components with different proportions was constructed. Two liquid crystalline areas were found with olive oil in different regions in the ternary diagram with two nonionic surfactants, called SLC1 (S1 and SLC2 (S2. These systems showed lamellar liquid crystals that remained stable during the entire analysis time. The systems were also characterized rheologically with pseudoplastic behavior without thixotropy. The texture and bioadhesion assays showed that formulations were similar statistically (p < 0.05, indicating that the increased amount of water in S2 did not interfere with the bioadhesive properties of the systems. In vitro cytotoxic assays showed that formulations did not present cytotoxicity. Olive oil-based systems may be a promising platform for skin delivery of drugs.

  9. PLGA-Based Nanoparticles: a Safe and Suitable Delivery Platform for Osteoarticular Pathologies.

    Science.gov (United States)

    Riffault, Mathieu; Six, Jean-Luc; Netter, Patrick; Gillet, Pierre; Grossin, Laurent

    2015-12-01

    Despite the promising applications of PLGA based particles, studies examining the fate and consequences of these particles after intra-articular administration in the joint are scanty. This study was carried out to evaluate the neutrality of the unloaded delivery system on different articular cell types. To facilitate tracking, we have thus developed a fluorescent core of particles, combined to a hyaluronate shell for cell recognition. Fluorescence pictures were taken at time intervals to assess the internalization and the corresponding inflammatory response was monitored by RT-qPCR and biochemical measurements. After NPs pre-treatment, mesenchymal stem cells (MSCs) were cultured into chondrogenic, adipogenic or osteogenic differentiation media, to investigate if NPs exposure interferes with differentiation ability. Finally, intra-articular injections were performed in healthy rat knees and joint's structure analysed by histological studies. Particles were detected in cytoplasm 8 h after exposure. Internalization led to a slight and reversible increase of inflammatory markers, but lower than in inflammatory conditions. We have confirmed particles exposure minimal neutrality on MSCs pluripotency. Histological exams of joint after intra-articular injections do not demonstrate any side effects of NPs. Our findings suggest that such a delivery platform is well tolerated locally and could be used to deliver active molecules to the joint.

  10. Preclinical development of drug delivery systems for paclitaxel-based cancer chemotherapy.

    Science.gov (United States)

    Wang, Feihu; Porter, Michael; Konstantopoulos, Alexandros; Zhang, Pengcheng; Cui, Honggang

    2017-12-10

    Paclitaxel (PTX) is one of the most successful drugs ever used in cancer chemotherapy, acting against a variety of cancer types. Formulating PTX with Cremophor EL and ethanol (Taxol®) realized its clinical potential, but the formulation falls short of expectations due to side effects such as peripheral neuropathy, hypotension, and hypersensitivity. Abraxane®, the albumin bound PTX, represents a superior replacement of Taxol® that mitigates the side effects associated with Cremophor EL. While Abraxane® is now considered a gold standard in chemotherapy, its 21% response rate leaves much room for further improvement. The quest for safer and more effective cancer treatments has led to the development of a plethora of innovative PTX formulations, many of which are currently undergoing clinical trials. In this context, we review recent development of PTX drug delivery systems and analyze the design principles underpinning each delivery strategy. We chose several representative examples to highlight the opportunities and challenges of polymeric systems, lipid-based formulations, as well as prodrug strategies. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Last Advances in Nanocarriers-Based Drug Delivery Systems for Colorectal Cancer.

    Science.gov (United States)

    Rama, Ana R; Jimenez-Lopez, Julia; Cabeza, Laura; Jimenez-Luna, Cristina; Leiva, Maria C; Perazzoli, Gloria; Hernandez, Rosa; Zafra, Inmaculada; Ortiz, Raul; Melguizo, Consolacion; Prados, Jose

    2016-01-01

    Colorectal cancer is the third most common type of cancer in both, men and women. The development of metastasis is very frequent, especially in patients with advanced stage, who require intensive chemotherapy that often results in poor response and significant morbidity. The undesirable effects of intensive chemotherapy on normal cells and the development of multidrug resistance are two of the main causes of treatment failure. Recent advances in nanotechnology allow to target cancer cells using cytotoxic drugs without affecting normal cells. Nanocarriers such as liposomes, polymeric nanoparticles and carbon nanotubes, among others, are able to improve drug distribution and bioavailability, cytotoxic concentration in the tumor mass and drug delivery to tumor tissue and, at the same time, reduce side effects. Current research studies are being conducted to develop new biomaterials that improve the characteristics of these nanomolecules. Several preclinical assays have disclosed the efficacy of nanotherapy in colon cancer, although further clinical trials will be necessary to demonstrate its efficacy. This review discusses the current status and the potential advantages of using nanocarrier-based drug delivery systems for colorectal cancer.

  12. A Multi Agent Based Organizational Architecture for Dynamic Pickup and Delivery Problem

    Directory of Open Access Journals (Sweden)

    Tahar Guerram

    2017-01-01

    Full Text Available Pickup and Delivery Problem (PDP consists of searching an optimal set of vehicles and an optimal set of routes, one route by each vehicle, in order to pickup items from a set of origins and deliver them to another set of destinations. Pickup and delivery problem is a class of complex systems whose complexity is NP Hard. In PDP real life applications, heuristics and meta heuristics methods are used in order to obtain an acceptable solution in reasonable execution time. When unpredictable events, like for example path cut and vehicles failure, may occur during the PDP schedule execution, we say that the PDP is dynamic (DPDP and in this case we have to revise this schedule. In this paper, we propose a multi agent architecture for DPDP based on an organizational architecture. Supported by a formal framework, the proposed architecture allows us to show, through a case study that computed solution for the studied problem could be done in a parallel manner which attenuates substantially the problem complexity.

  13. Ethosomes-based topical delivery system of antihistaminic drug for treatment of skin allergies.

    Science.gov (United States)

    Goindi, Shishu; Dhatt, Bhavnita; Kaur, Amanpreet

    2014-01-01

    Cetirizine is indicated for the treatment of allergic conditions such as insect bites and stings, atopic and contact dermatitis, eczema, urticaria. This investigation deals with development of a novel ethosome-based topical formulation of cetirizine dihydrochloride for effective delivery. The optimised formulation consisting of drug, phospholipon 90 G™ and ethanol was characterised for drug content, entrapment efficiency, pH, vesicular size, spreadability and rheological behaviour. The ex vivo permeation studies through mice skin showed highest permeation flux (16.300 ± 0.300 µg/h/cm(2)) and skin retention (20.686 ± 0.517 µg/cm(2)) for cetirizine-loaded ethosomal vesicles as compared to conventional formulations. The in vivo pharmacodynamic evaluation of optimised formulation was assessed against oxazolone-induced atopic dermatitis (AD) in mice. The parameters evaluated were reduction in scratching score, erythema score, skin hyperplasia and dermal eosinophil count. Our results suggest that ethosomes are effective carriers for dermal delivery of antihistaminic drug, cetirizine, for the treatment of AD.

  14. Synthesis of New Functionalized Citric Acid-based Dendrimers as Nanocarrier Agents for Drug Delivery

    Directory of Open Access Journals (Sweden)

    Sanaz Motamedi

    2011-06-01

    Full Text Available Introduction: Citric acid-polyethylene glycol-citric acid (CPEGC triblock dendrimers can serve as potential delivery systems. Methods: In this investigation, CPEGC triblock dendrimers were synthesized and then imidazole groups were conjugated onto the surface of the G1, G2 and G3 of the obtained dendrimers. In order to study the type of the interactions between the functionalized dendrimers and a drug molecule, Naproxen which contains acidic groups, was examined as a hydrophobic drug in which the interactions would be of the electrostatic kind between its acidic groups and the lone pair electrons of nitrogen atom in imidazole groups. The quantity of the trapped drug and also the amount of its release were measured with UV spectrometric method in pH 1, 7.4 and 10. The average diameter of the nanocarriers was measured by Dynamic Light Scattering (DLS technique Results: The size range of particles was determined to be 16-50 nm for different generations. The rate of the release increased in pH=10 in all generations due to the increase in Naproxen solubility and the hydrolysis of the esteric bonds in the mentioned pH. The results showed that the amount of the trapped drug increased with the increase in the generation of the dendrimer and pH. Conclusion: Based on our findings, we suggest CPEGC triblock dendrimers possess great potential to be used as drug/gene delivery system.

  15. The intracellular trafficking mechanism of Lipofectamine-based transfection reagents and its implication for gene delivery.

    Science.gov (United States)

    Cardarelli, Francesco; Digiacomo, Luca; Marchini, Cristina; Amici, Augusto; Salomone, Fabrizio; Fiume, Giuseppe; Rossetta, Alessandro; Gratton, Enrico; Pozzi, Daniela; Caracciolo, Giulio

    2016-05-11

    Lipofectamine reagents are widely accepted as "gold-standard" for the safe delivery of exogenous DNA or RNA into cells. Despite this, a satisfactory mechanism-based explanation of their superior efficacy has remained mostly elusive thus far. Here we apply a straightforward combination of live cell imaging, single-particle tracking microscopy, and quantitative transfection-efficiency assays on live cells to unveil the intracellular trafficking mechanism of Lipofectamine/DNA complexes. We find that Lipofectamine, contrary to alternative formulations, is able to efficiently avoid active intracellular transport along microtubules, and the subsequent entrapment and degradation of the payload within acidic/digestive lysosomal compartments. This result is achieved by random Brownian motion of Lipofectamine-containing vesicles within the cytoplasm. We demonstrate here that Brownian diffusion is an efficient route for Lipofectamine/DNA complexes to avoid metabolic degradation, thus leading to optimal transfection. By contrast, active transport along microtubules results in DNA degradation and subsequent poor transfection. Intracellular trafficking, endosomal escape and lysosomal degradation appear therefore as highly interdependent phenomena, in such a way that they should be viewed as a single barrier on the route for efficient transfection. As a matter of fact, they should be evaluated in their entirety for the development of optimized non-viral gene delivery vectors.

  16. Hydrolyzed polyacrylamide grafted maize starch based microbeads: application in pH responsive drug delivery.

    Science.gov (United States)

    Setty, C Mallikarjuna; Deshmukh, Anand S; Badiger, Aravind M

    2014-09-01

    The present study details the synthesis, characterization and pharmaceutical application of hydrolysed polyacrylamide grafted maize starch (HPam-g-MS) as promising polymeric material for the development of pH responsive microbeads. Different grades of graft copolymer were synthesized by changing the net microwave irradiation time, while keeping all other factors constant. Acute oral toxicity study performed in rodents ensured the bio-safety of graft copolymer for clinical application. Various batches of aceclofenac loaded microbeads were prepared by ionic gelation method using synthesized graft copolymers and evaluated for formulation parameters. FTIR spectroscopy confirmed the chemical compatibility between drug and graft copolymer. Results of in vitro release study (USP type-II) carried out in two different pH media (pH 1.2 acid buffer and pH 7.4 phosphate buffer) showed that release rate of drug from developed microbeads was a function of both: (a) surrounding pH and (b) the matrix composition. The drug release was relatively higher at alkaline pH as compared to acidic pH and this feature is desirable from viewpoint of site specific drug delivery. A direct correlation was observed between percentage grafting and microbeads performance and it presents a scope for further research on application and optimization of HPam-g-MS based microbeads as drug delivery carriers. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Project Delivery System Mode Decision Based on Uncertain AHP and Fuzzy Sets

    Science.gov (United States)

    Kaishan, Liu; Huimin, Li

    2017-12-01

    The project delivery system mode determines the contract pricing type, project management mode and the risk allocation among all participants. Different project delivery system modes have different characteristics and applicable scope. For the owners, the selection of the delivery mode is the key point to decide whether the project can achieve the expected benefits, it relates to the success or failure of project construction. Under the precondition of comprehensively considering the influence factors of the delivery mode, the model of project delivery system mode decision was set up on the basis of uncertain AHP and fuzzy sets, which can well consider the uncertainty and fuzziness when conducting the index evaluation and weight confirmation, so as to rapidly and effectively identify the most suitable delivery mode according to project characteristics. The effectiveness of the model has been verified via the actual case analysis in order to provide reference for the construction project delivery system mode.

  18. pH-responsive biocompatible fluorescent polymer nanoparticles based on phenylboronic acid for intracellular imaging and drug delivery.

    Science.gov (United States)

    Li, Shengliang; Hu, Kelei; Cao, Weipeng; Sun, Yun; Sheng, Wang; Li, Feng; Wu, Yan; Liang, Xing-Jie

    2014-11-21

    To address current medical challenges, there is an urgent need to develop drug delivery systems with multiple functions, such as simultaneous stimuli-responsive drug release and real-time imaging. Biocompatible polymers have great potential for constructing smart multifunctional drug-delivery systems through grafting with other functional ligands. More importantly, novel biocompatible polymers with intrinsic fluorescence emission can work as theranostic nanomedicines for real-time imaging and drug delivery. Herein, we developed a highly fluorescent nanoparticle based on a phenylboronic acid-modified poly(lactic acid)-poly(ethyleneimine)(PLA-PEI) copolymer loaded with doxorubicin (Dox) for intracellular imaging and pH-responsive drug delivery. The nanoparticles exhibited superior fluorescence properties, such as fluorescence stability, no blinking and excitation-dependent fluorescence behavior. The Dox-loaded fluorescent nanoparticles showed pH-responsive drug release and were more effective in suppressing the proliferation of MCF-7 cells. In addition, the biocompatible fluorescent nanoparticles could be used as a tool for intracellular imaging and drug delivery, and the process of endosomal escape was traced by real-time imaging. These pH-responsive and biocompatible fluorescent polymer nanoparticles, based on phenylboronic acid, are promising tools for intracellular imaging and drug delivery.

  19. Redox-Responsive Polysulfide-Based Biodegradable Organosilica Nanoparticles for Delivery of Bioactive Agents.

    Science.gov (United States)

    Hadipour Moghaddam, Seyyed Pouya; Saikia, Jiban; Yazdimamaghani, Mostafa; Ghandehari, Hamidreza

    2017-06-28

    Design and development of silica nanoparticles (SiO2 NPs) with a controlled degradation profile promises effective drug delivery with a predetermined carrier elimination profile. In this research, we fabricated a series of redox-responsive polysulfide-based biodegradable SiO2 NPs with low polydispersity and with variations in size (average diameters of 58 ± 7, 108 ± 11, 110 ± 9, 124 ± 9, and 332 ± 6 nm), porosity, and composition (disulfide vs tetrasulfide bonds). The degradation kinetics of the nanoparticles was analyzed in the presence of 8 mM glutathione (GSH), mimicking the intracellular reducing condition. Results indicate that porosity and core composition play the predominant roles in the degradation rate of these nanoparticles. The 108 nm mesoporous disulfide-based nanoparticles showed the highest degradation rate among all the synthesized nanoparticles. Transmission electron microscopy (TEM) reveals that nonporous nanoparticles undergo surface erosion, while porous nanoparticles undergo both surface and bulk erosion under reducing environment. The cytotoxicity of these nanoparticles in RAW 264.7 macrophages was evaluated. Results show that all these nanoparticles with the IC50 values ranging from 233 ± 42 to 705 ± 17 μg mL-1 do not have cytotoxic effect in macrophages at concentrations less than 125 μg mL-1. The degradation products of these nanoparticles collected within 15 days did not show cytotoxicity in the same macrophage cell line after 24 h of incubation. In vitro doxorubicin (DOX) release was examined in 108 nm mesoporous disulfide-based nanoparticles in the absence and presence of 8 mM GSH. It was shown that drug release depends on intracellular reducing conditions. Due to their ease of synthesis and scale up, robust structure, and the ability to control size, composition, release, and elimination, biodegradable SiO2 NPs provide an alternative platform for delivery of bioactive and imaging agents.

  20. Social Insurance for Delivery (Jampersal Policy in Indonesia: Culture-Based Approach for Improving Delivery by Health Workers in Rural Areas

    Directory of Open Access Journals (Sweden)

    Riswati Riswati

    2015-06-01

    the job training for midewife should be established. Conclusion:Culture-Based Approach has important role to improve Delivery By Health WorkersIn Rural Areas. Recommendation:Shorterm and longterm priority of policy should be set up.

  1. Novel therapeutic approaches for various cancer types using a modified sleeping beauty-based gene delivery system.

    Directory of Open Access Journals (Sweden)

    In-Sun Hong

    Full Text Available Successful gene therapy largely depends on the selective introduction of therapeutic genes into the appropriate target cancer cells. One of the most effective and promising approaches for targeting tumor tissue during gene delivery is the use of viral vectors, which allow for high efficiency gene delivery. However, the use of viral vectors is not without risks and safety concerns, such as toxicities, a host immune response towards the viral antigens or potential viral recombination into the host's chromosome; these risks limit the clinical application of viral vectors. The Sleeping Beauty (SB transposon-based system is an attractive, non-viral alternative to viral delivery systems. SB may be less immunogenic than the viral vector system due to its lack of viral sequences. The SB-based gene delivery system can stably integrate into the host cell genome to produce the therapeutic gene product over the lifetime of a cell. However, when compared to viral vectors, the non-viral SB-based gene delivery system still has limited therapeutic efficacy due to the lack of long-lasting gene expression potential and tumor cell specific gene transfer ability. These limitations could be overcome by modifying the SB system through the introduction of the hTERT promoter and the SV40 enhancer. In this study, a modified SB delivery system, under control of the hTERT promoter in conjunction with the SV40 enhancer, was able to successfully transfer the suicide gene (HSV-TK into multiple types of cancer cells. The modified SB transfected cancer cells exhibited a significantly increased cancer cell specific death rate. These data suggest that our modified SB-based gene delivery system can be used as a safe and efficient tool for cancer cell specific therapeutic gene transfer and stable long-term expression.

  2. Individually addressable multi-chamber electroporation platform with dielectrophoresis and alternating-current-electro-osmosis assisted cell positioning

    National Research Council Canada - National Science Library

    Park, Sinwook; Bassat, Dana Ben; Yossifon, Gilad

    2014-01-01

    ...) and alternating-current-electro-osmosis (ACEO) assisted cell/particle manipulation. A spatial gradient of electroporation parameters was generated within a microchamber array and validated using normal human dermal fibroblast (NHDF...

  3. Acute and Long-Term Effects of Full-Power Electroporation Ablation Directly on the Porcine Esophagus

    NARCIS (Netherlands)

    Neven, Kars; van Es, René; van Driel, Vincent; van Wessel, Harry; Fidder, Herma|info:eu-repo/dai/nl/274788187; Vink, Aryan; Doevendans, Pieter|info:eu-repo/dai/nl/164248366; Wittkampf, Fred|info:eu-repo/dai/nl/080434436

    BACKGROUND: Esophageal ulceration and fistula are complications of pulmonary vein isolation using thermal energy sources. Irreversible electroporation is a novel, nonthermal ablation modality for pulmonary vein isolation. A single 200 J application can create deep myocardial lesions. Acute and

  4. Relationship among reaction rate, release rate and efficiency of nanomachine-based targeted drug delivery.

    Science.gov (United States)

    Zhao, Qingying; Li, Min; Luo, Jun

    2017-12-04

    In nanomachine applications towards targeted drug delivery, drug molecules released by nanomachines propagate and chemically react with tumor cells in aqueous environment. If the nanomachines release drug molecules faster than the tumor cells react, it will result in loss and waste of drug molecules. It is a potential issue associated with the relationship among reaction rate, release rate and efficiency. This paper aims to investigate the relationship among reaction rate, release rate and efficiency based on two drug reception models. We expect to pave a way for designing a control method of drug release. We adopted two analytical methods that one is drug reception process based on collision with tumors and another is based on Michaelis Menten enzymatic kinetics. To evaluate the analytical formulations, we used the well-known simulation framework N3Sim to establish simulations. The analytical results of the relationship among reaction rate, release rate and efficiency is obtained, which match well with the numerical simulation results in a 3-D environment. Based upon two drug reception models, the results of this paper would be beneficial for designing a control method of nanomahine-based drug release.

  5. Gene transfer to pre-hematopoietic and committed hematopoietic precursors in the early mouse Yolk Sac: a comparative study between in situ electroporation and retroviral transduction

    Directory of Open Access Journals (Sweden)

    Lécluse Yann

    2007-07-01

    Full Text Available Abstract Background Hematopoietic development in vertebrate embryos results from the sequential contribution of two pools of precursors independently generated. While intra-embryonic precursors harbour the features of hematopoietic stem cells (HSC, precursors formed earlier in the yolk sac (YS display limited differentiation and self-renewal potentials. The mechanisms leading to the generation of the precursors in both sites are still largely unknown, as are the molecular basis underlying their different potential. A possible approach to assess the role of candidate genes is to transfer or modulate their expression/activity in both sites. We thus designed and compared transduction protocols to target either native extra-embryonic precursors, or hematopoietic precursors. Results One transduction protocol involves transient modification of gene expression through in situ electroporation of the prospective blood islands, which allows the evolution of transfected mesodermal cells in their "normal" environment, upon organ culture. Following in situ electroporation of a GFP reporter construct into the YS cavity of embryos at post-streak (mesodermal/pre-hematopoietic precursors or early somite (hematopoietic precursors stages, high GFP expression levels as well as a good preservation of cell viability is observed in YS explants. Moreover, the erythro-myeloid progeny typical of the YS arises from GFP+ mesodermal cells or hematopoietic precursors, even if the number of targeted precursors is low. The second approach, based on retroviral transduction allows a very efficient transduction of large precursor numbers, but may only be used to target 8 dpc YS hematopoietic precursors. Again, transduced cells generate a progeny quantitatively and qualitatively similar to that of control YS. Conclusion We thus provide two protocols whose combination may allow a thorough study of both early and late events of hematopoietic development in the murine YS. In situ

  6. Live demonstration: Screen printed, microwave based level sensor for automated drug delivery

    KAUST Repository

    Karimi, Muhammad Akram

    2018-01-02

    Level sensors find numerous applications in many industries to automate the processes involving chemicals. Recently, some commercial ultrasound based level sensors are also being used to automate the drug delivery process [1]. Some of the most desirable features of level sensors to be used for medical use are their non-intrusiveness, low cost and consistent performance. In this demo, we will present a completely new method of sensing the liquid level using microwaves. It is a common stereotype to consider microwaves sensing mechanism as being expensive. Unlike usual expensive, intrusive and bulky microwave methods of level sensing using guided radars, we will present an extremely low cost printed, non-intrusive microwave sensor to reliably sense the liquid level.

  7. Cell-based delivery of glucagon-like peptide-1 using encapsulated mesenchymal stem cells

    DEFF Research Database (Denmark)

    Wallrapp, Christine; Thoenes, Eric; Thürmer, Frank

    2013-01-01

    Glucagon-like peptide-1 (GLP-1) CellBeads are cell-based implants for the sustained local delivery of bioactive factors. They consist of GLP-1 secreting mesenchymal stem cells encapsulated in a spherically shaped immuno-isolating alginate matrix. A highly standardized and reproducible encapsulation...... and quality control is performed in compliance with good manufacturing practice and fulfils all regulatory requirements for human clinical use. GLP-1 CellBeads combine the neuro- and cardioprotective properties of both GLP-1 and mesenchymal stem cells. First promising results were obtained from preclinical...... method is described for the manufacturing of homogeneous CellBeads. Viability and sustained secretion was shown for the recombinant GLP-1 and the cell endogenous bioactive factors like vascular endothelial growth factor, neurotrophin 3 (NT-3) and glial cell line-derived neurotrophic factor. Manufacturing...

  8. Fabrication and Analysis of Tapered Tip Silicon Microneedles for MEMS based Drug Delivery System

    Directory of Open Access Journals (Sweden)

    Muhammad Waseem Ashraf

    2010-11-01

    Full Text Available In this paper, a novel design of transdermal drug delivery (TDD system is presented. The proposed system consists of controlled electronic circuit and microelectromechanical system (MEMS based devices like microneedles, micropump, flow sensor, and blood pressure sensor. The aim of this project is to develop a system that can eliminate the limitations associated with oral therapy. In this phase tapered tip silicon microneedles have been fabricated using inductively coupled plasma (ICP etching technology. Using ANSYS, simulation of microneedles has been conducted before the fabrication process to test the design suitability for TDD. More over multifield analysis of reservoir integrated with microneedle array using piezoelectric actuator has also been performed. The effects of frequency and voltage on actuator and fluid flow rate through 6×6 microneedle array have been investigated. This work provides envisage data to design suitable devices for TDD.

  9. Microspheres made of poly(epsilon-caprolactone)-based amphiphilic copolymers: potential in sustained delivery of proteins.

    Science.gov (United States)

    Quaglia, Fabiana; Ostacolo, Luisanna; Nese, Giuseppe; De Rosa, Giuseppe; La Rotonda, Maria Immacolata; Palumbo, Rosario; Maglio, Giovanni

    2005-10-20

    Microspheres of amphiphilic multi-block poly(ester-ether)s (PEE)s and poly(ester-ether-amide)s (PEEA)s based on poly(epsilon-caprolactone) (PCL) were investigated as delivery systems for proteins. The interest was mainly focused on the effect of their molecular structure and composition on the overall properties of the microspheres, encapsulating bovine serum albumin (BSA) as a model protein. PEEs and PEEAs were prepared using a alpha,omega-dihydroxy-terminated PCL macromer (Mn= 2.0 kDa) as a hydrophobic component. Hydrophilic oxyethylene sequences were generated using poly(ethylene oxide)s (PEO)s of different molecular mass (Mn= 300-600 Da) in the case of PEEs, or 4,7,10-trioxa-1,13-tridecanediamine (Trioxy) and PEO150 (Mn= 150 Da) in the case of PEEAs. The copolymers showed a decrease of Tm and crystallinity values as compared with PCL. Within each class of copolymers, the bulk hydrophilicity increased with increasing the number of oxyethylene groups in the chain repeat unit. PEEAs were more hydrophilic than PEEs with a similar number of oxyethylene groups. Discrete spherical particles were prepared by both PEEs and PEEAs and their BSA encapsulation efficiency related to copolymer properties. Interestingly, the insertion of short hydrophilic segments is enough to significantly affect protein distribution inside microspheres and its release profiles, as compared to PCL microspheres. Different degradation rates and mechanisms were observed for copolymer microspheres, mainly depending on the distribution of oxyethylene units along the chain. The results highlight that a fine control over the structural parameters of amphiphilic PCL-based multi-block copolymers is a key factor for their application in the field of protein delivery.

  10. Wearable/disposable sweat-based glucose monitoring device with multistage transdermal drug delivery module

    Science.gov (United States)

    Lee, Hyunjae; Song, Changyeong; Hong, Yong Seok; Kim, Min Sung; Cho, Hye Rim; Kang, Taegyu; Shin, Kwangsoo; Choi, Seung Hong; Hyeon, Taeghwan; Kim, Dae-Hyeong

    2017-01-01

    Electrochemical analysis of sweat using soft bioelectronics on human skin provides a new route for noninvasive glucose monitoring without painful blood collection. However, sweat-based glucose sensing still faces many challenges, such as difficulty in sweat collection, activity variation of glucose oxidase due to lactic acid secretion and ambient temperature changes, and delamination of the enzyme when exposed to mechanical friction and skin deformation. Precise point-of-care therapy in response to the measured glucose levels is still very challenging. We present a wearable/disposable sweat-based glucose monitoring device integrated with a feedback transdermal drug delivery module. Careful multilayer patch design and miniaturization of sensors increase the efficiency of the sweat collection and sensing process. Multimodal glucose sensing, as well as its real-time correction based on pH, temperature, and humidity measurements, maximizes the accuracy of the sensing. The minimal layout design of the same sensors also enables a strip-type disposable device. Drugs for the feedback transdermal therapy are loaded on two different temperature-responsive phase change nanoparticles. These nanoparticles are embedded in hyaluronic acid hydrogel microneedles, which are additionally coated with phase change materials. This enables multistage, spatially patterned, and precisely controlled drug release in response to the patient’s glucose level. The system provides a novel closed-loop solution for the noninvasive sweat-based management of diabetes mellitus. PMID:28345030

  11. Multicoil resonance-based parallel array for smart wireless power delivery.

    Science.gov (United States)

    Mirbozorgi, S A; Sawan, M; Gosselin, B

    2013-01-01

    This paper presents a novel resonance-based multicoil structure as a smart power surface to wirelessly power up apparatus like mobile, animal headstage, implanted devices, etc. The proposed powering system is based on a 4-coil resonance-based inductive link, the resonance coil of which is formed by an array of several paralleled coils as a smart power transmitter. The power transmitter employs simple circuit connections and includes only one power driver circuit per multicoil resonance-based array, which enables higher power transfer efficiency and power delivery to the load. The power transmitted by the driver circuit is proportional to the load seen by the individual coil in the array. Thus, the transmitted power scales with respect to the load of the electric/electronic system to power up, and does not divide equally over every parallel coils that form the array. Instead, only the loaded coils of the parallel array transmit significant part of total transmitted power to the receiver. Such adaptive behavior enables superior power, size and cost efficiency then other solutions since it does not need to use complex detection circuitry to find the location of the load. The performance of the proposed structure is verified by measurement results. Natural load detection and covering 4 times bigger area than conventional topologies with a power transfer efficiency of 55% are the novelties of presented paper.

  12. Nanoemulsion-based electrolyte triggered in situ gel for ocular delivery of acetazolamide.

    Science.gov (United States)

    Morsi, Nadia; Ibrahim, Magdy; Refai, Hanan; El Sorogy, Heba

    2017-06-15

    In the present work the antiglaucoma drug, acetazolamide, was formulated as an ion induced nanoemulsion-based in situ gel for ocular delivery aiming a sustained drug release and an improved therapeutic efficacy. Different acetazolamide loaded nanoemulsion formulations were prepared using peanut oil, tween 80 and/or cremophor EL as surfactant in addition to transcutol P or propylene glycol as cosurfactant. Based on physicochemical characterization, the nanoemulsion formulation containing mixed surfactants and transcutol P was selected to be incorporated into ion induced in situ gelling systems composed of gellan gum alone and in combination with xanthan gum, HPMC or carbopol. The nanoemulsion based in situ gels showed a significantly sustained drug release in comparison to the nanoemulsion. Gellan/xanthan and gellan/HPMC possessed good stability at all studied temperatures, but gellan/carbopol showed partial drug precipitation upon storage and was therefore excluded from the study. Gellan/xanthan and gellan/HPMC showed higher therapeutic efficacy and more prolonged intraocular pressure lowering effect relative to that of commercial eye drops and oral tablet. Gellan/xanthan showed superiority over gellan/HPMC in all studied parameters and is thus considered as a promising mucoadhesive nanoemulsion-based ion induced in situ gelling formula for topical administration of acetazolamide. Copyright © 2017. Published by Elsevier B.V.

  13. Formulation and characterization of lipid-based drug delivery system of raloxifene-microemulsion and self-microemulsifying drug delivery system

    Directory of Open Access Journals (Sweden)

    Hetal Thakkar

    2011-01-01

    Full Text Available Background : Raloxifene, a second-generation selective estrogen receptor modulator (SERM used to prevent osteoporosis in postmenopausal women is administered orally in the form of a tablet. The absolute bioavailability of the drug is only 2% because of extensive hepatic first-pass metabolism. Lipid-based formulations are reported to reduce the first-pass metabolism by promoting its lymphatic uptake. Materials and Methods : In the present investigation, microemulsion and Self-Microemulsifying Drug Delivery System (SMEDDS formulations of Raloxifene were prepared. The prepared formulations were characterized for drug loading, size, transparency, zeta potential, Transmission Electron Microscopy (TEM and in vitro intestinal permeability. Results : The results indicated that high drug loading, optimum size and desired zeta potential and transparency could be achieved with both SMEDDS and microemulsion. The TEM studies indicated the absence of aggregation with both the systems. The in vitro intestinal permeability results showed that the permeation of the drug from the microemulsion and SMEDDs was significantly higher than that obtained from the drug dispersion and marketed formulation. Conclusion : Lipid based formulations such as microemulsion and Self Microemulsifying drug delivery systems are expected to increase the oral bioavailability as evidenced by the increased intestinal permeation.

  14. Simulation and Analysis of Microneedle for Drug Delivery based on Structural and Fluid Flow Mechanics

    OpenAIRE

    Neha Mane; Ashok Gaikwad

    2013-01-01

    MEMS (Micro Electro Mechanical System) design systems are used for the development of microneedles, which are used for drug delivery. The drug delivery through micronnedle is painless which is not in case of transdermal needles. In this paper the simulation and analysis of microneedle is presented. The typical structure of a microneedle is simulated for studies of the structural behavior of microneedle and the fluid mechanics of the drug delivery. The studies show that the microneedle with si...

  15. 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies

    CERN Document Server

    Kramar, Peter

    2016-01-01

    This volume presents the proceedings of the 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies (WC2015). The congress took place in Portorož, Slovenia, during the week of September 6th to 10th, 2015. The scientific part of the Congress covered different aspects of electroporation and related technologies and included the following main topics:   ·         Application of pulsed electric fields technology in food: challenges and opportunities ·         Electrical impedance measurement for assessment of electroporation yield ·         Electrochemistry and electroporation ·         Electroporation meets electrostimulation ·         Electrotechnologies for food and biomass treatment ·         Food and biotechnology applications ·         In vitro electroporation - basic mechanisms ·         Interfacial behaviour of lipid-assemblies, membranes and cells in electric f...

  16. HPMA-based block copolymers promote differential drug delivery kinetics for hydrophobic and amphiphilic molecules.

    Science.gov (United States)

    Tomcin, Stephanie; Kelsch, Annette; Staff, Roland H; Landfester, Katharina; Zentel, Rudolf; Mailänder, Volker

    2016-04-15

    We describe a method how polymeric nanoparticles stabilized with (2-hydroxypropyl)methacrylamide (HPMA)-based block copolymers are used as drug delivery systems for a fast release of hydrophobic and a controlled release of an amphiphilic molecule. The versatile method of the miniemulsion solvent-evaporation technique was used to prepare polystyrene (PS) as well as poly-d/l-lactide (PDLLA) nanoparticles. Covalently bound or physically adsorbed fluorescent dyes labeled the particles' core and their block copolymer corona. Confocal laser scanning microscopy (CLSM) in combination with flow cytometry measurements were applied to demonstrate the burst release of a fluorescent hydrophobic drug model without the necessity of nanoparticle uptake. In addition, CLSM studies and quantitative calculations using the image processing program Volocity® show the intracellular detachment of the amphiphilic block copolymer from the particles' core after uptake. Our findings offer the possibility to combine the advantages of a fast release for hydrophobic and a controlled release for an amphiphilic molecule therefore pointing to the possibility to a 'multi-step and multi-site' targeting by one nanocarrier. We describe thoroughly how different components of a nanocarrier end up in cells. This enables different cargos of a nanocarrier having a consecutive release and delivery of distinct components. Most interestingly we demonstrate individual kinetics of distinct components of such a system: first the release of a fluorescent hydrophobic drug model at contact with the cell membrane without the necessity of nanoparticle uptake. Secondly, the intracellular detachment of the amphiphilic block copolymer from the particles' core after uptake occurs. This offers the possibility to combine the advantages of a fast release for a hydrophobic substance at the time of interaction of the nanoparticle with the cell surface and a controlled release for an amphiphilic molecule later on therefore

  17. SU-F-T-289: MLC Fluence Sonogram Based Delivery Quality Assurance for Bilateral Breast Irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Thiyagarajan, Rajesh; Karrthick, KP; Kataria, Tejinder; Mahendran, Ramu; Selvan, Tamil; Duraikannu, Palani [Division of Radiation Oncology, Medanta The Medicity, Gurgaon, Haryana (India); Raj, Nambi [Department of Physics, School of Advanced sciences, VIT University, Vellore (India); Arunai, N

    2016-06-15

    Purpose: Performing DQA for Bilateral (B-L) breast tomotherapy is a challenging task due to the limitation of any commercially available detector array or film. Aim of this study is to perform DQA for B-L breast tomotherapy plan using MLC fluence sinogram. Methods: Treatment plan was generated on Tomotherapy system for B-L breast tumour. B-L breast targets were given 50.4 Gy prescribed over 28 fractions. Plan is generated with 6 MV photon beam & pitch was set to 0.3. As the width of the total target is 39 cm (left & right) length is 20 cm. DQA plan delivered without any phantom on the mega voltage computed tomography (MCVT) detector system. The pulses recorded by MVCT system were exported to the delivery analysis software (Tomotherapy Inc.) for reconstruction. The detector signals are reconstructed to a sonogram and converted to MLC fluence sonogram. The MLC fluence sinogram compared with the planned fluence sinogram. Also point dose measured with cheese phantom and ionization chamber to verify the absolute dose component Results: Planned fluence sinogram and reconstructed MLC fluence sinogram were compared using Gamma metric. MLC positional difference and intensity of the beamlet were used as parameters to evaluate gamma. 3 mm positional difference and 3% beamlet intensity difference were used set for gamma calculation. A total of 26784 non-zero beamlets were included in the analysis out of which 161 beamlets had gamma more than 1. The gamma passing rate found to be 99.4%. Point dose measurements were within 1.3% of the calculated dose. Conclusion: MLC fluence sinogram based delivery quality assurance performed for bilateral breast irradiation. This would be a suitable alternate for large volume targets like bilateral breast, Total body irradiation etc. However conventional method of DQA should be used to validate this method periodically.

  18. Small field dose delivery evaluations using cone beam optical computed tomography-based polymer gel dosimetry

    Directory of Open Access Journals (Sweden)

    Timothy Olding

    2011-01-01

    Full Text Available This paper explores the combination of cone beam optical computed tomography with an N-isopropylacrylamide (NIPAM-based polymer gel dosimeter for three-dimensional dose imaging of small field deliveries. Initial investigations indicate that cone beam optical imaging of polymer gels is complicated by scattered stray light perturbation. This can lead to significant dosimetry failures in comparison to dose readout by magnetic resonance imaging (MRI. For example, only 60% of the voxels from an optical CT dose readout of a 1 l dosimeter passed a two-dimensional Low′s gamma test (at a 3%, 3 mm criteria, relative to a treatment plan for a well-characterized pencil beam delivery. When the same dosimeter was probed by MRI, a 93% pass rate was observed. The optical dose measurement was improved after modifications to the dosimeter preparation, matching its performance with the imaging capabilities of the scanner. With the new dosimeter preparation, 99.7% of the optical CT voxels passed a Low′s gamma test at the 3%, 3 mm criteria and 92.7% at a 2%, 2 mm criteria. The fitted interjar dose responses of a small sample set of modified dosimeters prepared (a from the same gel batch and (b from different gel batches prepared on the same day were found to be in agreement to within 3.6% and 3.8%, respectively, over the full dose range. Without drawing any statistical conclusions, this experiment gives a preliminary indication that intrabatch or interbatch NIPAM dosimeters prepared on the same day should be suitable for dose sensitivity calibration.

  19. SU-E-T-789: Validation of 3DVH Accuracy On Quantifying Delivery Errors Based On Clinical Relevant DVH Metrics

    Energy Technology Data Exchange (ETDEWEB)

    Ma, T; Kumaraswamy, L [Roswell Park Cancer Institute, Buffalo, NY (United States)

    2015-06-15

    Purpose: Detection of treatment delivery errors is important in radiation therapy. However, accurate quantification of delivery errors is also of great importance. This study aims to evaluate the 3DVH software’s ability to accurately quantify delivery errors. Methods: Three VMAT plans (prostate, H&N and brain) were randomly chosen for this study. First, we evaluated whether delivery errors could be detected by gamma evaluation. Conventional per-beam IMRT QA was performed with the ArcCHECK diode detector for the original plans and for the following modified plans: (1) induced dose difference error up to ±4.0% and (2) control point (CP) deletion (3 to 10 CPs were deleted) (3) gantry angle shift error (3 degree uniformly shift). 2D and 3D gamma evaluation were performed for all plans through SNC Patient and 3DVH, respectively. Subsequently, we investigated the accuracy of 3DVH analysis for all cases. This part evaluated, using the Eclipse TPS plans as standard, whether 3DVH accurately can model the changes in clinically relevant metrics caused by the delivery errors. Results: 2D evaluation seemed to be more sensitive to delivery errors. The average differences between ECLIPSE predicted and 3DVH results for each pair of specific DVH constraints were within 2% for all three types of error-induced treatment plans, illustrating the fact that 3DVH is fairly accurate in quantifying the delivery errors. Another interesting observation was that even though the gamma pass rates for the error plans are high, the DVHs showed significant differences between original plan and error-induced plans in both Eclipse and 3DVH analysis. Conclusion: The 3DVH software is shown to accurately quantify the error in delivered dose based on clinically relevant DVH metrics, where a conventional gamma based pre-treatment QA might not necessarily detect.

  20. SOA-Based Model for Value-Added ITS Services Delivery

    Directory of Open Access Journals (Sweden)

    Luis Felipe Herrera-Quintero

    2014-01-01

    Full Text Available Integration is currently a key factor in intelligent transportation systems (ITS, especially because of the ever increasing service demands originating from the ITS industry and ITS users. The current ITS landscape is made up of multiple technologies that are tightly coupled, and its interoperability is extremely low, which limits ITS services generation. Given this fact, novel information technologies (IT based on the service-oriented architecture (SOA paradigm have begun to introduce new ways to address this problem. The SOA paradigm allows the construction of loosely coupled distributed systems that can help to integrate the heterogeneous systems that are part of ITS. In this paper, we focus on developing an SOA-based model for integrating information technologies (IT into ITS to achieve ITS service delivery. To develop our model, the ITS technologies and services involved were identified, catalogued, and decoupled. In doing so, we applied our SOA-based model to integrate all of the ITS technologies and services, ranging from the lowest-level technical components, such as roadside unit as a service (RSUAAS, to the most abstract ITS services that will be offered to ITS users (value-added services. To validate our model, a functionality case study that included all of the components of our model was designed.

  1. Volume-Based Enteral Nutrition Support Regimen Improves Caloric Delivery but May Not Affect Clinical Outcomes in Critically Ill Patients.

    Science.gov (United States)

    Haskins, Ivy N; Baginsky, Mary; Gamsky, Nathaniel; Sedghi, Kia; Yi, Sojung; Amdur, Richard L; Gergely, Mary; Sarani, Babak

    2017-05-01

    Meeting enteral nutrition goals is an ongoing challenge in the intensive care unit (ICU). Most hospitals use rate-based (RB) protocols for nutrient delivery. Previous studies have found that volume-based (VB) protocols improve delivery of prescribed calories. However, these studies did not assess clinical outcomes. We hypothesize that a VB method will improve the delivery of prescribed calories and lead to improved clinical outcomes. A before-and-after study was performed following implementation of a VB feeding protocol in an adult mixed medical-surgical ICU. Formal institutional review board approval was obtained. The effect of RB and VB protocols on percentage of goal calories received, ICU length of stay (LOS), hospital LOS, mortality, days on the ventilator, and rates of infection were investigated using the Kruskal-Wallis test of differences. Multivariate regression was used to identify independent predictors of outcome. Significance was defined as P power to corroborate these findings.

  2. Design and Characterization of a Silk-Fibroin-Based Drug Delivery Platform Using Naproxen as a Model Drug

    Directory of Open Access Journals (Sweden)

    Tatyana Dyakonov

    2012-01-01

    Full Text Available The objective of this proof-of-concept study was to develop a platform for controlled drug delivery based on silk fibroin (SF and to explore the feasibility of using SF in oral drug delivery. The SF-containing matrixes were prepared via spray-drying and film casting, and the release profile of the model drug naproxen sodium was evaluated. Attenuated total reflectance Fourier transform infrared spectroscopy (FTIR has been used to observe conformational changes in SF- and drug-containing compositions. SF-based films, spray-dried microparticles, and matrixes loaded with naproxen were prepared. Both FTIR spectra and in vitro dissolution data demonstrated that SF β-sheet conformation regulates the release profile of naproxen. The controlled release characteristics of the SF-containing compositions were evaluated as a function of SF concentration, temperature, and exposure to dehydrating solvents. The results suggest that SF may be an attractive polymer for use in controlled drug delivery systems.

  3. Viral and nonviral delivery systems for gene delivery

    Directory of Open Access Journals (Sweden)

    Nouri Nayerossadat

    2012-01-01

    Full Text Available Gene therapy is the process of introducing foreign genomic materials into host cells to elicit a therapeutic benefit. Although initially the main focus of gene therapy was on special genetic disorders, now diverse diseases with different patterns of inheritance and acquired diseases are targets of gene therapy. There are 2 major categories of gene therapy, including germline gene therapy and somatic gene therapy. Although germline gene therapy may have great potential, because it is currently ethically forbidden, it cannot be used; however, to date human gene therapy has been limited to somatic cells. Although numerous viral and nonviral gene delivery systems have been developed in the last 3 decades, no delivery system has been designed that can be applied in gene therapy of all kinds of cell types in vitro and in vivo with no limitation and side effects. In this review we explain about the history of gene therapy, all types of gene delivery systems for germline (nuclei, egg cells, embryonic stem cells, pronuclear, microinjection, sperm cells and somatic cells by viral [retroviral, adenoviral, adeno association, helper-dependent adenoviral systems, hybrid adenoviral systems, herpes simplex, pox virus, lentivirus, Epstein-Barr virus] and nonviral systems (physical: Naked DNA, DNA bombardant, electroporation, hydrodynamic, ultrasound, magnetofection and (chemical: Cationic lipids, different cationic polymers, lipid polymers. In addition to the above-mentioned, advantages, disadvantages, and practical use of each system are discussed.

  4. Light-sensitive lipid-based nanoparticles for drug delivery: design principles and future considerations for biological applications.

    Science.gov (United States)

    Yavlovich, Amichai; Smith, Brandon; Gupta, Kshitij; Blumenthal, Robert; Puri, Anu

    2010-10-01

    Radiation-based therapies aided by nanoparticles have been developed for decades, and can be primarily categorized into two main platforms. First, delivery of payload of photo-reactive drugs (photosensitizers) using the conventional nanoparticles, and second, design and development of photo-triggerable nanoparticles (primarily liposomes) to attain light-assisted on-demand drug delivery. The main focus of this review is to provide an update of the history, current status and future applications of photo-triggerable lipid-based nanoparticles (light-sensitive liposomes). We will begin with a brief overview on the applications of liposomes for delivery of photosensitizers, including the choice of photosensitizers for photodynamic therapy, as well as the currently available light sources (lasers) used for these applications. The main segment of this review will encompass the details of strategies used to develop photo-triggerable liposomes for their drug delivery function. The principles underlying the assembly of photoreactive lipids into nanoparticles (liposomes) and photo-triggering mechanisms will be presented. We will also discuss factors that limit the applications of these liposomes for in vivo triggered drug delivery and emerging concepts that may lead to the biologically viable photo-activation strategies. We will conclude with our view point on the future perspectives of light-sensitive liposomes in the clinic.

  5. Small nanosized poly(vinyl benzyl trimethylammonium chloride) based polyplexes for siRNA delivery

    NARCIS (Netherlands)

    Lou, Bo|info:eu-repo/dai/nl/37052537X; Beztsinna, Nataliia|info:eu-repo/dai/nl/411277928; Mountrichas, Grigoris; van den Dikkenberg, Joep B.|info:eu-repo/dai/nl/304841420; Pispas, Stergios; Hennink, Wim E.|info:eu-repo/dai/nl/070880409

    2017-01-01

    The success of siRNA gene therapy requires the availability of safe and efficient delivery systems. In the present study, we investigated poly(vinyl benzyl trimethylammonium chloride) (PVTC) and its block copolymer with poly(oligo(ethyleneglycol) methacrylate) (POEGMA) as delivery vector for siRNA.

  6. Menu-based service access and delivery pattern: Towards achieving equatable access to digital services

    CSIR Research Space (South Africa)

    Makitla, I

    2014-11-01

    Full Text Available . This capability is used in basic service such as Short Message Service (SMS) as well as Unstructured Supplementary Service Data (USSD). There is a need for a service access and delivery pattern that can facilitate the delivery of services using common...

  7. Towards an Innovative Web-Based Lab Delivery System for a Management Information Systems Course

    Science.gov (United States)

    Breimer, Eric; Cotler, Jami; Yoder, Robert

    2011-01-01

    While online systems are an essential component of distance learning, they can also play a critical role in improving the delivery of activities in a traditional laboratory setting. The quality and effectiveness of online course delivery is often compared to equivalent face-to-face alternatives. In our approach, we have harnessed what we feel to…

  8. Abort or retry - A role for legal knowledge based systems in electronic service delivery?

    NARCIS (Netherlands)

    Leenes, R.E.; Wimmer, Maria A.

    2003-01-01

    Electronic service delivery is closely tied to legal decision making. Legal decision making is by nature complicated. It involves the (strict) application of rules, but it also inevitably leaves room for discretion. If the ambition of electronic service delivery is taken seriously, this means that

  9. [Establishment of animal model for electroporation-mediated gene therapy in distraction osteogenesis of rabbit mandible].

    Science.gov (United States)

    Wu, Guo-Ping; Li, Sheng-Hua; Li, De-Ping; Yang, Zhi-Hui; He, Xiao-Chuan; Liao, Yi; Guo, Li

    2009-07-01

    To evaluate the feasibility of electroporation-mediated transfection of recombinant plasmid to mandibular distraction area of rabbit in vivo. New-Zeland rabbit were employed. The mandible was distracted 3 days after operation at a rate of 0.8 mm per day for 7 days. The rabbits were randomly divided into 3 groups as group A (recombinant plasmid pIRES-VEGF165-EGFP), group B (recombinant plasmid plRES-VEGF165-EGFP) and group C (normal saline). The rabbits were sacrified at 3 hours, 1, 3, 7 and 14 d after injection respectively. The tissue at the distraction area was taken out for frozen section. The gene expression was assessed by the detection of expression of green fluorescence protein (GFP) using fluorescence microscope. The liver and kidney function test (ALT, AST, BUN, Scr) and the histological examination of heart, liver and kidney were also performed. GFP was seen in the distraction area in group A and group B 3 hours after injection, which increased at the 1st day, reached peak value at the 3rd day, decreased at the 7th day and was very lower at 14th day. The GFP expression was much stronger in group A than in group B. GFP was not expressed in group C. There was no statistical difference in the concentration of ALT, AST, BUN and Scr in serum of rabbits among the three groups. Electroporation-mediated transfection of recombinant plasmid can be expressed in the distraction area of rabbits, and there was no toxicity to the liver and kidney of rabbits. Electroporation could obviously improve transfection efficiency in vivo. It indicates that electroporation-mediated transfection of recombinant plasmid to distraction area tissue of rabbits is feasible.

  10. Irreversible Electroporation of a Hepatocellular Carcinoma Lesion Adjacent to a Transjugular Intrahepatic Portosystemic Shunt Stent Graft

    Energy Technology Data Exchange (ETDEWEB)

    Niessen, Christoph; Jung, Ernst Michael; Wohlgemuth, Walter A. [Department of Radiology, University Medical Center Regensburg, Regensburg D-93053 (Germany); Trabold, Benedikt [Department of Anaesthesia, University Medical Center Regensburg, Regensburg D-93053 (Germany); Haimerl, Michael; Schreyer, Andreas; Stroszczynski, Christian; Wiggermann, Philipp [Department of Radiology, University Medical Center Regensburg, Regensburg D-93053 (Germany)

    2013-07-01

    We report in a 65-year-old man hepatocellular carcinoma adjacent to a transjugular intrahepatic portosystemic shunt stent-graft which was successfully treated with irreversible electroporation (IRE). IRE is a new non-thermal tissue ablation technique which uses electrical pulses to induce cell necrosis by irreversible membrane poration. IRE proved to be more advantageous in the ablation of perivascular tumor with little injury to the surrounding structures.

  11. The systematic study of the electroporation and electrofusion of B16-F1 and CHO cells in isotonic and hypotonic buffer.

    Science.gov (United States)

    Usaj, Marko; Kanduser, Masa

    2012-09-01

    The fusogenic state of the cell membrane can be induced by external electric field. When two fusogenic membranes are in close contact, cell fusion takes place. An appropriate hypotonic treatment of cells before the application of electric pulses significantly improves electrofusion efficiency. How hypotonic treatment improves electrofusion is still not known in detail. Our results indicate that at given induced transmembrane potential electroporation was not affected by buffer osmolarity. In contrast to electroporation, cells' response to hypotonic treatment significantly affects their electrofusion. High fusion yield was observed when B16-F1 cells were used; this cell line in hypotonic buffer resulted in 41 ± 9 % yield, while in isotonic buffer 32 ± 11 % yield was observed. Based on our knowledge, these fusion yields determined in situ by dual-color fluorescence microscopy are among the highest in electrofusion research field. The use of hypotonic buffer was more crucial for electrofusion of CHO cells; the fusion yield increased from below 1 % in isotonic buffer to 10 ± 4 % in hypotonic buffer. Since the same degree of cell permeabilization was achieved in both buffers, these results indicate that hypotonic treatment significantly improves fusion yield. The effect could be attributed to improved physical contact of cell membranes or to enhanced fusogenic state of the cell membrane itself.

  12. Human serum albumin-based nanoparticle-mediated in vitro gene delivery.

    Directory of Open Access Journals (Sweden)

    Monica Langiu

    Full Text Available The genetic treatment of neurodegenerative diseases still remains a challenging task since many approaches fail to deliver the therapeutic material in relevant concentrations into the brain. As viral vectors comprise the risk of immune and inflammatory responses, human serum albumin (HSA nanoparticles were found to represent a safer and more convenient alternative. Their ability to cross the blood-brain barrier (BBB and deliver drugs into the brain in order to enhance gene-based therapy has been previously demonstrated. The present study deals with the development of pGL3-PEI-coated HSA nanoparticles and subsequent in vitro testing in cerebellar granular and HeLa cells. The luciferase control vector pGL3 was chosen as reporter plasmid encoding for the firefly luciferase protein, linear polyethylenimine (22 kDa as endosomolytic agent for enhancing the cells' transfection. Studies on particle characteristics, their cellular uptake into aforementioned cell lines and on subcellular localisation, and transfection efficiency in the cerebellar cells proved the feasibility of nanoparticle-based gene delivery.

  13. An extended EPQ-based problem with a discontinuous delivery policy, scrap rate, and random breakdown.

    Science.gov (United States)

    Chiu, Singa Wang; Lin, Hong-Dar; Song, Ming-Syuan; Chen, Hsin-Mei; Chiu, Yuan-Shyi P

    2015-01-01

    In real supply chain environments, the discontinuous multidelivery policy is often used when finished products need to be transported to retailers or customers outside the production units. To address this real-life production-shipment situation, this study extends recent work using an economic production quantity- (EPQ-) based inventory model with a continuous inventory issuing policy, defective items, and machine breakdown by incorporating a multiple delivery policy into the model to replace the continuous policy and investigates the effect on the optimal run time decision for this specific EPQ model. Next, we further expand the scope of the problem to combine the retailer's stock holding cost into our study. This enhanced EPQ-based model can be used to reflect the situation found in contemporary manufacturing firms in which finished products are delivered to the producer's own retail stores and stocked there for sale. A second model is developed and studied. With the help of mathematical modeling and optimization techniques, the optimal run times that minimize the expected total system costs comprising costs incurred in production units, transportation, and retail stores are derived, for both models. Numerical examples are provided to demonstrate the applicability of our research results.

  14. A Nano-in-Nano Polymer-Dendrimer Nanoparticle-Based Nanosystem for Controlled Multidrug Delivery.

    Science.gov (United States)

    Zhao, Zongmin; Lou, Song; Hu, Yun; Zhu, Jie; Zhang, Chenming

    2017-08-07

    Codelivery of multiple chemotherapeutics with different action mechanisms is a promising strategy for cancer treatment. In this study, we developed a novel polymer-dendrimer hybrid nanoparticle-based nanosystem for efficient and controlled codelivery of two model chemotherapeutics, doxorubicin (DOX) and paclitaxel (PTX). The nanosystem was characterized to have a nano-in-nano structure with a size of around 150 nm. The model drugs could feasibly be loaded into the nanosystem ratiometrically with high drug-loading contents by controlling the feeding drug ratios. Also, the model drugs could be released from the nanosystem following a sequential release manner-specifically, quick PTX release and sustained DOX release. Acidic pH was found to enhance the release of both drugs. Moreover, the nanosystem was taken up by cancer cells rapidly and efficiently, and the delivered drugs could release sustainably and efficiently in cells to reach their action targets. In vitro cytotoxicity results demonstrated that, by optimizing drug ratios, the dual-drug-loaded nanosystem could result in better antitumor efficacy than the single-drug-loaded nanosystem or free dual-drug combination. Furthermore, the dual-drug-loaded nanosystem could induce significant changes in both the nucleus and tubulin patterns synergistically. All data suggest that the nano-in-nano polymer-dendrimer hybrid nanoparticle-based nanosystem is a promising candidate to achieve controlled multidrug delivery for effective combination cancer therapy.

  15. Advanced review of graphene-based nanomaterials in drug delivery systems: Synthesis, modification, toxicity and application.

    Science.gov (United States)

    Zhang, Qi; Wu, Zhuona; Li, Ning; Pu, Yiqiong; Wang, Bing; Zhang, Tong; Tao, Jiansheng

    2017-08-01

    The discovery of graphene, a notable achievement in the field of novel carbon nanomaterials, has triggered the worldwide exploration of the biomedical applications of this material since 2004 because of its unique properties. The two-dimensional planar structure, large surface area, chemical stability, mechanical stability, and good biocompatibility of graphene are promising for applications in drug delivery systems (DDSs). In this review, we briefly discuss the characteristics, synthesis, and modification of graphene. We also investigate its toxicity and its applications in DDSs, with several representative examples. This review presents a comprehensive summary of graphene-based nanomaterials from their characteristics to their synthesis and applications, as well as their in vitro and in vivo evaluation in medicine. This paper provides a guiding strategy for the selection of optimal approaches to the fabrication of nanocarriers that are suitable for medical treatments and to controlling the toxicity within therapeutic safety limits. The promising achievements made with graphene-based nanomaterials indicate several possibilities for further biomedical research, as well as theoretical and applied development. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Design and optimization of resonance-based efficient wireless power delivery systems for biomedical implants.

    Science.gov (United States)

    Ramrakhyani, A K; Mirabbasi, S; Mu Chiao

    2011-02-01

    Resonance-based wireless power delivery is an efficient technique to transfer power over a relatively long distance. This technique typically uses four coils as opposed to two coils used in conventional inductive links. In the four-coil system, the adverse effects of a low coupling coefficient between primary and secondary coils are compensated by using high-quality (Q) factor coils, and the efficiency of the system is improved. Unlike its two-coil counterpart, the efficiency profile of the power transfer is not a monotonically decreasing function of the operating distance and is less sensitive to changes in the distance between the primary and secondary coils. A four-coil energy transfer system can be optimized to provide maximum efficiency at a given operating distance. We have analyzed the four-coil energy transfer systems and outlined the effect of design parameters on power-transfer efficiency. Design steps to obtain the efficient power-transfer system are presented and a design example is provided. A proof-of-concept prototype system is implemented and confirms the validity of the proposed analysis and design techniques. In the prototype system, for a power-link frequency of 700 kHz and a coil distance range of 10 to 20 mm, using a 22-mm diameter implantable coil resonance-based system shows a power-transfer efficiency of more than 80% with an enhanced operating range compared to ~40% efficiency achieved by a conventional two-coil system.

  17. Molecular Dynamics Study of Stability and Diffusion of Graphene-Based Drug Delivery Systems

    Directory of Open Access Journals (Sweden)

    Xiunan Wang

    2015-01-01

    Full Text Available Graphene, a two-dimensional nanomaterial with unique biomedical properties, has attracted great attention due to its potential applications in graphene-based drug delivery systems (DDS. In this work graphene sheets with various sizes and graphene oxide functionalized with polyethylene glycol (GO-PEG are utilized as nanocarriers to load anticancer drug molecules including CE6, DOX, MTX, and SN38. We carried out molecular dynamics calculations to explore the energetic stabilities and diffusion behaviors of the complex systems with focuses on the effects of the sizes and functionalization of graphene sheets as well as the number and types of drug molecules. Our study shows that the binding of graphene-drug complex is favorable when the drug molecules and finite graphene sheets become comparable in sizes. The boundaries of finite sized graphene sheets restrict the movement of drug molecules. The double-side loading often slows down the diffusion of drug molecules compared with the single-side loading. The drug molecules bind more strongly with GO-PEG than with pristine graphene sheets, demonstrating the advantages of functionalization in improving the stability and biocompatibility of graphene-based DDS.

  18. Microsponge based drug delivery system for augmented gastroparesis therapy: Formulation development and evaluation

    Directory of Open Access Journals (Sweden)

    Riyaz Ali M. Osmani

    2015-10-01

    Full Text Available The intention behind the present work was to develop a microsponge based novel dosage form for sustained delivery of domperidone. Quasi-emulsion solvent diffusion method was employed using Eudragit RS-100 with various drug–polymer ratios for the preparation of microsponges. For optimization purposes, several factors which affect microparticles' physical properties were investigated. Characterization techniques followed for the formed microsponges were DSC, FTIR, SEM, XRD and particle size analysis, along with morphology, drug loading and in vitro drug release. It was found that there were no chemical interactions between drugs and polymers used as per DSC and FTIR results. The drug–polymer ratio showed remarkable impact on drug content, encapsulation efficiency and particle size. SEM micrographs revealed that microsponges were spherical in shape with porous surface, and had 104 ± 0.22 µm mean particle size. The microsponges were then loaded in capsules followed by in vitro drug release study; which depicted that microsponges with drug–polymer ratio of 1:2 were more proficient to give extended drug release of 76.38% at the end of 8 h, superior in contrast to conventional marketed formulation Domstal®, which got exhausted incredibly earlier by releasing 82.57% drug at the end of ½ h only. Hence, the developed microsponge based formulation of domperidone would be an expectant, promising substitute to conventional therapy of gastroparesis, emesis and alike gastric ailments.

  19. Transdermal delivery of alprazolam from a monolithic patch: formulation based on in vitro characterization.

    Science.gov (United States)

    Soler, L I; Boix, A; Lauroba, J; Colom, H; Domenech, J

    2012-10-01

    Alprazolam, a benzodiazepine widely used for the treatment of psychiatric disorders, has been aimed to be formulated in a transdermal delivery system (TDS) prototype. A series of TDS prototypes dosed in all cases at 0.35 mg·cm(-2) of alprazolam were prepared as a monolithic drug in adhesive matrix using acrylic pressure-sensitive adhesives (PSA) of acrylate vinyl acetate (Duro-tack(®)). The effects of several permeation enhancers as azone, transcutol, propylene glycol, dodecyl alcohol, decyl alcohol, diethanolamine, N-methyl pyrrolidone and lauric acid were studied. Prototypes have been characterized based on adhesion parameters (peel adhesion and shear adhesion), in vitro human skin permeation and in vitro drug release according to European Pharmacopoeia for the selected prototype. Best results show that a combination of permeation enhancers from different chemical groups is able to provide almost a 33 fold increase in the transdermal alprazolam flux of an aqueous saturated dispersion (from 0.054 ± 0.019 to 1.76 ± 0.21 μg h.cm(-2)). Based on these in vitro flux data, a predictive simulation of the achievable plasmatic levels was performed assuming a constant systemic infusion of drug. In summary, it is possible to obtain a prototype of a TDS of alprazolam with adequate adhesive properties (peel adhesion and shear adhesion) and able to predict sustained therapeutic plasmatic levels.

  20. Priority-Based Content Delivery in the Internet of Vehicles through Named Data Networking

    Directory of Open Access Journals (Sweden)

    Marica Amadeo

    2016-11-01

    Full Text Available Named Data Networking (NDN has been recently proposed as a prominent solution for content delivery in the Internet of Vehicles (IoV, where cars equipped with a variety of wireless communication technologies exchange information aimed to support safety, traffic efficiency, monitoring and infotainment applications. The main NDN tenets, i.e., name-based communication and in-network caching, perfectly fit the demands of time- and spatially-relevant content requested by vehicles regardless of their provenance. However, existing vehicular NDN solutions have not been targeted to wisely ensure prioritized traffic treatment based on the specific needs of heterogeneous IoV content types. In this work, we propose a holistic NDN solution that, according to the demands of data traffic codified in NDN content names, dynamically shapes the NDN forwarding decisions to ensure the appropriate prioritization. Specifically, our proposal first selects the outgoing interface(s (i.e., 802.11, LTE for NDN packets and then properly tunes the timing of the actual transmissions. Simulation results show that the proposed enhancements succeed in achieving differentiated traffic treatment, while keeping traffic load under control.

  1. Tuning structural parameters for the optimization of drug delivery performance of cyclodextrin-based nanosponges.

    Science.gov (United States)

    Venuti, Valentina; Rossi, Barbara; Mele, Andrea; Melone, Lucio; Punta, Carlo; Majolino, Domenico; Masciovecchio, Claudio; Caldera, Fabrizio; Trotta, Francesco

    2017-03-01

    In light of the recent development of new soft materials, nanostructured self-assembled systems have attracted attention in a variety of technological fields of high social impact. Cyclodextrin nanosponges (CDNS) represent a new and highly versatile class of cross-linked cyclodextrin (CD)-based nanoporous polymers. Their intriguing properties, including safety, biodegradability, negligible toxicity, marked swelling behavior, superior inclusion capability with respect to native CD, are the bases for potential for applications in drug delivery, tissue engineering and regenerative medicine. Areas covered: We report on the state-of-art concerning a detailed characterization of structural and dynamical features of CDNS explored by the combined use of different and complementary techniques, such as Fourier transform infrared absorption in attenuated total reflectance geometry (FTIR-ATR) and Raman spectroscopies, and high resolution magic angle spinning (HR-MAS) NMR spectroscopy. The ambitious objective is to furnish an exhaustive survey of the role played by hydrophobic and hydrophilic groups within the cross-linked network, in dry and swollen states, in determining the macroscopic functional features of CDNS. Expert opinion: The reported results may significantly contribute in the rational design and optimization of new stimuli-responsive systems exhibiting tunable inclusion/release properties, adapted to the therapeutic demands of pathology.

  2. An Extended EPQ-Based Problem with a Discontinuous Delivery Policy, Scrap Rate, and Random Breakdown

    Directory of Open Access Journals (Sweden)

    Singa Wang Chiu

    2015-01-01

    Full Text Available In real supply chain environments, the discontinuous multidelivery policy is often used when finished products need to be transported to retailers or customers outside the production units. To address this real-life production-shipment situation, this study extends recent work using an economic production quantity- (EPQ- based inventory model with a continuous inventory issuing policy, defective items, and machine breakdown by incorporating a multiple delivery policy into the model to replace the continuous policy and investigates the effect on the optimal run time decision for this specific EPQ model. Next, we further expand the scope of the problem to combine the retailer’s stock holding cost into our study. This enhanced EPQ-based model can be used to reflect the situation found in contemporary manufacturing firms in which finished products are delivered to the producer’s own retail stores and stocked there for sale. A second model is developed and studied. With the help of mathematical modeling and optimization techniques, the optimal run times that minimize the expected total system costs comprising costs incurred in production units, transportation, and retail stores are derived, for both models. Numerical examples are provided to demonstrate the applicability of our research results.

  3. Improved electroporation procedure for genetic transformation of Dekkera/Brettanomyces bruxellensis.

    Science.gov (United States)

    Miklenić, Marina; Žunar, Bojan; Štafa, Anamarija; Svetec, Ivan-Krešimir

    2015-12-01

    Yeast Dekkera/Brettanomyces bruxellensis is one of the most common contaminants in wine industry, but also one of the most promising candidates for large-scale bioethanol production. Brettanomyces bruxellensis not only produces and tolerates high ethanol concentrations, but can also ferment cellobiose and adapt to lignocellulose hydrolasate. Furthermore, genome sequences of several B. bruxellensis strains are available, and efforts have been made to develop tools for genetic transformation of this yeast. Previously, we reported a successful transformation using lithium acetate/PEG method and electroporation, however, with very low transformation efficiency (10-20 transformants μg(-1)). Here we describe an optimization of electroporation procedure which resulted in a significant increase of transformation efficiency (2.8 × 10(3) transformants μg(-1)). Several key transformation parameters were optimized including cell growth phase, density of cells in the transformation sample and electroporation settings. We determined that treating the cells with both lithium acetate (100 mM) and dithiothreitol (35 mM) synergistically improves transformation efficiency. Using the described procedure around 500 transformants can be obtained per transformation sample with 180 ng of non-homologous linear transforming fragment. Additionally, several transformants were obtained with less than 1 ng of DNA demonstrating that this procedure is adequate even when very limited amount of DNA is available. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Effects of deformability and thermal motion of lipid membrane on electroporation: By molecular dynamics simulations

    KAUST Repository

    Sun, Sheng

    2011-01-01

    Effects of mechanical properties and thermal motion of POPE lipid membrane on electroporation were studied by molecular dynamics simulations. Among simulations in which specific atoms of lipids were artificially constrained at their equilibrium positions using a spring with force constant of 2.0kcal/(molÅ2) in the external electric field of 1.4kcal/(molÅe), only constraint on lateral motions of lipid tails prohibited electroporation while non-tail parts had little effects. When force constant decreased to 0.2kcal/(molÅ2) in the position constraints on lipid tails in the external electric field of 2.0kcal/(molÅe), water molecules began to enter the membrane. Position constraints of lipid tails allow water to penetrate from both sides of membrane. Thermal motion of lipids can induce initial defects in the hydrophobic core of membrane, which are favorable nucleation sites for electroporation. Simulations at different temperatures revealed that as the temperature increases, the time taken to the initial pore formation will decrease. © 2010 Elsevier Inc.

  5. Optimization of ectopic gene expression in skeletal muscle through DNA transfer by electroporation

    Directory of Open Access Journals (Sweden)

    Latour Mickey

    2004-05-01

    Full Text Available Abstract Background Electroporation (EP is a widely used non-viral gene transfer method. We have attempted to develop an exact protocol to maximize DNA expression while minimizing tissue damage following EP of skeletal muscle in vivo. Specifically, we investigated the effects of varying injection techniques, electrode surface geometry, and plasmid mediums. Results We found that as the amount of damage increased in skeletal muscle in response to EP, the level of β-galactosidase (β-gal expression drastically decreased and that there was no evidence of β-gal expression in damaged fibers. Two specific types of electrodes yielded the greatest amount of expression. We also discovered that DNA uptake in skeletal muscle following intra-arterial injection of DNA was significantly enhanced by EP. Finally, we found that DMSO and LipoFECTAMINE™, common enhancers of DNA electroporation in vitro, had no positive effect on DNA electroporation in vivo. Conclusions When injecting DNA intramuscularly, a flat plate electrode without any plasmid enhancers is the best method to achieve high levels of gene expression.

  6. Polycation-Based Gene Delivery: on the mechanisms of transfection with lactosylated and poly(ehtylene glycol) derivatized cationic polymers

    NARCIS (Netherlands)

    D.T. Klink

    2004-01-01

    textabstractGene delivery and therapy in general The concept of gene therapy is a promising approach towards clinical treatment of pathological processes. The underlying principle of gene therapy is based on the introduction of genetic material into living cells in order to achieve a therapeutic

  7. Internet-Based Intervention Training for Parents of Young Children with Disabilities: A Promising Service-Delivery Model

    Science.gov (United States)

    Meadan, Hedda; Daczewitz, Marcus E.

    2015-01-01

    Efficient early intervention (EI) services are required to serve the needs of young children with disabilities and the needs of their families. Effective EI includes family-centred practices, evidence-based interventions, parent involvement/training, and delivery in children's natural environments. Due to the challenges of providing…

  8. Effect of radiotherapy and hyperthermia on the tumor accumulation of HPMA copolymer-based drug delivery systems.

    Science.gov (United States)

    Lammers, Twan; Peschke, Peter; Kühnlein, Rainer; Subr, Vladimir; Ulbrich, Karel; Debus, Jürgen; Huber, Peter; Hennink, Wim; Storm, Gert

    2007-02-26

    Copolymers of N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic and well-characterized polymeric drug carriers that have been broadly implemented in the delivery of anticancer therapeutics. In an attempt to improve the tumor accumulation of HPMA copolymer-based drug delivery systems, their in vivo application was combined with radiotherapy and hyperthermia. As the effects of radiotherapy and hyperthermia were considered to depend significantly on the tumor model used, we first analyzed the accumulation of two differently sized HPMA copolymers in three different types of tumors, based on the syngeneic Dunning rat prostate carcinoma model. Subsequently, in these three models, the effects of different doses of radiotherapy and hyperthermia on the tumor accumulation of 31 kDa poly(HPMA), 65 kDa poly(HPMA) and 28 kDa poly(HPMA)-GFLG-doxorubicin were evaluated. It was found that the polymeric drug delivery systems accumulated effectively in all three tumor models. In addition, as opposed to hyperthermia, radiotherapy was found to improve the concentrations of the copolymers independent of the tumor model used. Based on these findings, we conclude that radiotherapy is an effective means for increasing the tumor accumulation of (polymeric) drug delivery systems, and we propose that the combination of carrier-based chemotherapy with radiotherapy holds significant potential for improving the treatment of advanced solid malignancies.

  9. Gold nanocluster-based vaccines for dual-delivery of antigens and immunostimulatory oligonucleotides

    Science.gov (United States)

    Tao, Yu; Zhang, Yan; Ju, Enguo; Ren, Hui; Ren, Jinsong

    2015-07-01

    We here report a facile one-pot synthesis of fluorescent gold nanoclusters (AuNCs) via the peptide biomineralization method, which can elicit specific immunological responses. The as-prepared peptide-protected AuNCs (peptide-AuNCs) display strong red fluorescence, and more importantly, as compared to the peptide alone, the immune stimulatory ability of the resulting peptide-AuNCs can not only be retained, but can also be efficaciously enhanced. Moreover, through a dual-delivery of antigen peptides and cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (ODNs), the as-prepared peptide-AuNC-CpG conjugates can also act as smart self-vaccines to assist in the generation of high immunostimulatory activity, and be applied as a probe for intracellular imaging. Both in vitro and in vivo studies provide strong evidence that the AuNC-based vaccines may be utilized as safe and efficient immunostimulatory agents that are able to prevent and/or treat a variety of ailments.We here report a facile one-pot synthesis of fluorescent gold nanoclusters (AuNCs) via the peptide biomineralization method, which can elicit specific immunological responses. The as-prepared peptide-protected AuNCs (peptide-AuNCs) display strong red fluorescence, and more importantly, as compared to the peptide alone, the immune stimulatory ability of the resulting peptide-AuNCs can not only be retained, but can also be efficaciously enhanced. Moreover, through a dual-delivery of antigen peptides and cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (ODNs), the as-prepared peptide-AuNC-CpG conjugates can also act as smart self-vaccines to assist in the generation of high immunostimulatory activity, and be applied as a probe for intracellular imaging. Both in vitro and in vivo studies provide strong evidence that the AuNC-based vaccines may be utilized as safe and efficient immunostimulatory agents that are able to prevent and/or treat a variety of ailments. Electronic supplementary information (ESI

  10. A novel method for sub-arc VMAT dose delivery verification based on portal dosimetry with an EPID.

    Science.gov (United States)

    Cools, Ruud A M; Dirkx, Maarten L P; Heijmen, Ben J M

    2017-11-01

    The EPID-based sub-arc verification of VMAT dose delivery requires synchronization of the acquired electronic portal images (EPIs) with the VMAT delivery, that is, establishment of the start- and stop-MU of the acquired images. To realize this, published synchronization methods propose the use of logging features of the linac or dedicated hardware solutions. In this study, we developed a novel, software-based synchronization method that only uses information inherently available in the acquired images. The EPIs are continuously acquired during pretreatment VMAT delivery and converted into Portal Dose Images (PDIs). Sub-arcs of approximately 10 MU are then defined by combining groups of sequentially acquired PDIs. The start- and stop-MUs of measured sub-arcs are established in a synchronization procedure, using only dosimetric information in measured and predicted PDIs. Sub-arc verification of a VMAT dose delivery is based on comparison of measured sub-arc PDIs with synchronized, predicted sub-arc PDIs, using γ-analyses. To assess the accuracy of this new method, measured and predicted PDIs were compared for 20 clinically applied VMAT prostate cancer plans. The sensitivity of the method for detection of delivery errors was investigated using VMAT deliveries with intentionally inserted, small perturbations (25 error scenarios; leaf gap deviations ≤ 1.5 mm, leaf motion stops during ≤ 15 MU, linac output error ≤ 2%). For the 20 plans, the average failed pixel rates (FPR) for full-arc and sub-arc dose QA were 0.36% ± 0.26% (1 SD) and 0.64% ± 0.88%, based on 2%/2 mm and 3%/3 mm γ-analyses, respectively. Small systematic perturbations of up to 1% output error and 1 mm leaf offset were detected using full-arc QA. Sub-arc QA was able to detect positioning errors in three leaves only during approximately 20 MU and small dose delivery errors during approximately 40 MU. In an ROC analysis, the area under the curve (AUC) for the combined full-arc/sub-arc approach was

  11. Enhanced immunogenicity of an HIV-1 DNA vaccine delivered with electroporation via combined intramuscular and intradermal routes.

    Science.gov (United States)

    Mann, Jamie F S; McKay, Paul F; Fiserova, Anezka; Klein, Katja; Cope, Alethea; Rogers, Paul; Swales, Julie; Seaman, Michael S; Combadiere, Behazine; Shattock, Robin J

    2014-06-01

    It is accepted that an effective prophylactic HIV-1 vaccine is likely to have the greatest impact on viral transmission rates. As previous reports have implicated DNA-priming, protein boost regimens to be efficient activators of humoral responses, we sought to optimize this regimen to further augment vaccine immunogenicity. Here we evaluated single versus concurrent intradermal (i.d.) and intramuscular (i.m.) vaccinations as a DNA-priming strategy for their abilities to elicit humoral and cellular responses against a model HIV-1 vaccine antigen, CN54-gp140. To further augment vaccine-elicited T and B cell responses, we enhanced cellular transfection with electroporation and then boosted the DNA-primed responses with homologous protein delivered subcutaneously (s.c.), intranasally (i.n.), i.m., or transcutaneously (t.c.). In mice, the concurrent priming regimen resulted in significantly elevated gamma interferon T cell responses and high-avidity antigen-specific IgG B cell responses, a hallmark of B cell maturation. Protein boosting of the concurrent DNA strategy further enhanced IgG concentrations but had little impact on T cell reactivity. Interestingly protein boosting by the subcutaneous route increased antibody avidity to a greater extent than protein boosting by either the i.m., i.n., or t.c. route, suggesting that this route may be preferential for driving B cell maturation. Using an alternative and larger animal model, the rabbit, we found the concurrent DNA-priming strategy followed by s.c. protein boosting to again be capable of eliciting high-avidity humoral responses and to also be able to neutralize HIV-1 pseudoviruses from diverse clades (clades A, B, and C). Taken together, we show that concurrent multiple-route DNA vaccinations induce strong cellular immunity, in addition to potent and high-avidity humoral immune responses. The route of vaccination has profound effects on prevailing immune responses. Due to the insufficient immunogenicity and

  12. School-based vaccination programmes: a systematic review of the evidence on organisation and delivery in high income countries.

    Science.gov (United States)

    Perman, Sarah; Turner, Simon; Ramsay, Angus I G; Baim-Lance, Abigail; Utley, Martin; Fulop, Naomi J

    2017-03-14

    Many countries have recently expanded their childhood immunisation programmes. Schools are an increasingly attractive setting for delivery of these new immunisations because of their ability to reach large numbers of children in a short period of time. However, there are organisational challenges to delivery of large-scale vaccination programmes in schools. Understanding the facilitators and barriers is important for improving the delivery of future school-based vaccination programmes. We undertook a systematic review of evidence on school-based vaccination programmes in order to understand the influence of organisational factors on the delivery of programmes. Our eligibility criteria were studies that (1) focused on childhood or adolescent vaccination programmes delivered in schools; (2) considered organisational factors that influenced the preparation or delivery of programmes; (3) were conducted in a developed or high-income country; and (4) had been peer reviewed. We searched for articles published in English between 2000 and 2015 using MEDLINE and HMIC electronic databases. Additional studies were identified by searching the Cochrane Library and bibliographies. We extracted data from the studies, assessed quality and the risk of bias, and categorised findings using a thematic framework of eight organisational factors. We found that most of the recent published literature is from the United States and is concerned with the delivery of pandemic or seasonal flu vaccination programmes at a regional (state) or local level. We found that the literature is largely descriptive and not informed by the use of theory. Despite this, we identified common factors that influence the implementation of programmes. These factors included programme leadership and governance, organisational models and institutional relationships, workforce capacity and roles particularly concerning the school nurse, communication with parents and students, including methods for obtaining consent

  13. School-based vaccination programmes: a systematic review of the evidence on organisation and delivery in high income countries

    Directory of Open Access Journals (Sweden)

    Sarah Perman

    2017-03-01

    Full Text Available Abstract Background Many countries have recently expanded their childhood immunisation programmes. Schools are an increasingly attractive setting for delivery of these new immunisations because of their ability to reach large numbers of children in a short period of time. However, there are organisational challenges to delivery of large-scale vaccination programmes in schools. Understanding the facilitators and barriers is important for improving the delivery of future school-based vaccination programmes. Methods We undertook a systematic review of evidence on school-based vaccination programmes in order to understand the influence of organisational factors on the delivery of programmes. Our eligibility criteria were studies that (1 focused on childhood or adolescent vaccination programmes delivered in schools; (2 considered organisational factors that influenced the preparation or delivery of programmes; (3 were conducted in a developed or high-income country; and (4 had been peer reviewed. We searched for articles published in English between 2000 and 2015 using MEDLINE and HMIC electronic databases. Additional studies were identified by searching the Cochrane Library and bibliographies. We extracted data from the studies, assessed quality and the risk of bias, and categorised findings using a thematic framework of eight organisational factors. Results We found that most of the recent published literature is from the United States and is concerned with the delivery of pandemic or seasonal flu vaccination programmes at a regional (state or local level. We found that the literature is largely descriptive and not informed by the use of theory. Despite this, we identified common factors that influence the implementation of programmes. These factors included programme leadership and governance, organisational models and institutional relationships, workforce capacity and roles particularly concerning the school nurse, communication with parents and

  14. Endocrine surgery as a model for value-based health care delivery.

    Science.gov (United States)

    Abdulla, Amer G; Ituarte, Philip H G; Wiggins, Randi; Teisberg, Elizabeth O; Harari, Avital; Yeh, Michael W

    2012-01-01

    Experts advocate restructuring health care in the United States into a value-based system that maximizes positive health outcomes achieved per dollar spent. We describe how a value-based system implemented by the University of California, Los Angeles UCLA Section of Endocrine Surgery (SES) has optimized both quality and costs while increasing patient volume. Two SES clinical pathways were studied, one allocating patients to the most appropriate surgical care setting based on clinical complexity, and another standardizing initial management of papillary thyroid carcinoma (PTC). The mean cost per endocrine case performed from 2005 to 2010 was determined at each of three care settings: A tertiary care inpatient facility, a community inpatient facility, and an ambulatory facility. Blood tumor marker levels (thyroglobulin, Tg) and reoperation rates were compared between PTC patients who underwent routine central neck dissection (CND) and those who did not. Surgical patient volume and regional market share were analyzed over time. The cost of care was substantially lower in both the community inpatient facility (14% cost savings) and the ambulatory facility (58% cost savings) in comparison with the tertiary care inpatient facility. Patients who underwent CND had lower Tg levels (6.6 vs 15.0 ng/mL; P = 0.024) and a reduced need for re-operation (1.5 vs 6.1%; P = 0.004) compared with those who did not undergo CND. UCLA maintained its position as the market leader in endocrine procedures while expanding its market share by 151% from 4.9% in 2003 to 7.4% in 2010. A value-driven health care delivery system can deliver improved clinical outcomes while reducing costs within a subspecialty surgical service. Broader application of these principles may contribute to resolving current dilemmas in the provision of care nationally.

  15. Antiretroviral agents and acid-base balance at delivery of the neonate

    Directory of Open Access Journals (Sweden)

    P. El-Beitune

    2007-07-01

    Full Text Available Limited evidence is available regarding antiretroviral (ARV safety for uninfected infants exposed to these drugs in utero. Our objective was to determine if ARV administered to pregnant women is associated with decreasing umbilical arterial pH and base excess in uninfected infants. A prospective study was conducted on 57 neonates divided into three groups: ZDV group, born to mothers taking zidovudine (N = 20, triple therapy (TT group, born to mothers taking zidovudine + lamivudine + nelfinavir (N = 25, and control group (N = 12, born to uninfected mothers. Umbilical cord blood was used to determine umbilical artery gases. A test was performed to calculate the sample by comparing means by the unpaired one-tailed t-test, with a = 0.05 and ß = 20%, indicating the need for a sample of 18 newborn infants for the study groups to detect differences higher than 20%. The control and ARV groups were similar in gestational age, birth weight, and Apgar scores. Values of pH, pCO2, bicarbonate, and base excess in cord arterial blood obtained at delivery from the newborns exposed to TT were 7.23, 43.2 mmHg, 19.5 mEq/L, and -8.5 nmol/L, respectively, with no significant difference compared to the control and ZDV groups. We conclude that intrauterine exposure to ARV is not associated with a pathological decrease in umbilical arterial pH or base excess. While our data are reassuring, follow-up is still limited and needs to be continued into adulthood because of the possible potential for adverse effects of triple antiretroviral agents.

  16. Oral Cholera Vaccination Delivery Cost in Low- and Middle-Income Countries: An Analysis Based on Systematic Review.

    Science.gov (United States)

    Mogasale, Vittal; Ramani, Enusa; Wee, Hyeseung; Kim, Jerome H

    2016-12-01

    Use of the oral cholera vaccine (OCV) is a vital short-term strategy to control cholera in endemic areas with poor water and sanitation infrastructure. Identifying, estimating, and categorizing the delivery costs of OCV campaigns are useful in analyzing cost-effectiveness, understanding vaccine affordability, and in planning and decision making by program managers and policy makers. To review and re-estimate oral cholera vaccination program costs and propose a new standardized categorization that can help in collation, analysis, and comparison of delivery costs across countries. Peer reviewed publications listed in PubMed database, Google Scholar and World Health Organization (WHO) websites and unpublished data from organizations involved in oral cholera vaccination. The publications and reports containing oral cholera vaccination delivery costs, conducted in low- and middle-income countries based on World Bank Classification. Limits are humans and publication date before December 31st, 2014. No participants are involved, only costs are collected. Oral cholera vaccination and cost estimation. A systematic review was conducted using pre-defined inclusion and exclusion criteria. Cost items were categorized into four main cost groups: vaccination program preparation, vaccine administration, adverse events following immunization and vaccine procurement; the first three groups constituting the vaccine delivery costs. The costs were re-estimated in 2014 US dollars (US$) and in international dollar (I$). Ten studies were identified and included in the analysis. The vaccine delivery costs ranged from US$0.36 to US$ 6.32 (in US$2014) which was equivalent to I$ 0.99 to I$ 16.81 (in I$2014). The vaccine procurement costs ranged from US$ 0.29 to US$ 29.70 (in US$2014), which was equivalent to I$ 0.72 to I$ 78.96 (in I$2014). The delivery costs in routine immunization systems were lowest from US$ 0.36 (in US$2014) equivalent to I$ 0.99 (in I$2014). The reported cost categories

  17. Synthesis and characterization of psyllium-NVP based drug delivery system through radiation crosslinking polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Baljit [Department of Chemistry, Himachal Pradesh University, Shimla 171005 (India)], E-mail: baljitsinghhpu@yahoo.com; Kumar, S. [Department of Chemistry, Himachal Pradesh University, Shimla 171005 (India)

    2008-08-15

    In order to develop the hydrogels meant for the drug delivery, we have prepared psyllium-N-vinylpyrrolidone (NVP) based hydrogels by radiation induced crosslinking. Polymers were characterized with SEMs, FTIR and swelling studies. Swelling of the hydrogels was studied as a function of monomer concentration, total radiation dose, temperature, pH and [NaCl] of the swelling medium. The swelling kinetics of the hydrogels and release dynamics of anticancer model drug (5-fluorouracil) from the hydrogels have been carried out for the evaluation of swelling and drug release mechanism. It has been observed that diffusion exponent 'n' have 0.8, 0.9, 0.8 and gel characteristics constant 'k' have 9.22 x 10{sup -3}, 2.06 x 10{sup -3}, 11.72 x 10{sup -3} values for the release of drug from the drug loaded hydrogels in distilled water, pH 2.2 buffer and pH 7.4 buffer, respectively. The present study shows that the release of drug from the hydrogels occurred through Non-Fickian diffusion mechanism.

  18. Synthesis and characterization of psyllium-NVP based drug delivery system through radiation crosslinking polymerization

    Science.gov (United States)

    Singh, Baljit; Kumar, S.

    2008-08-01

    In order to develop the hydrogels meant for the drug delivery, we have prepared psyllium- N-vinylpyrrolidone (NVP) based hydrogels by radiation induced crosslinking. Polymers were characterized with SEMs, FTIR and swelling studies. Swelling of the hydrogels was studied as a function of monomer concentration, total radiation dose, temperature, pH and [NaCl] of the swelling medium. The swelling kinetics of the hydrogels and release dynamics of anticancer model drug (5-fluorouracil) from the hydrogels have been carried out for the evaluation of swelling and drug release mechanism. It has been observed that diffusion exponent ' n' have 0.8, 0.9, 0.8 and gel characteristics constant ' k' have 9.22 × 10 -3, 2.06 × 10 -3, 11.72 × 10 -3 values for the release of drug from the drug loaded hydrogels in distilled water, pH 2.2 buffer and pH 7.4 buffer, respectively. The present study shows that the release of drug from the hydrogels occurred through Non-Fickian diffusion mechanism.

  19. Thermosensitive In Situ Gel Based on Solid Dispersion for Rectal Delivery of Ibuprofen.

    Science.gov (United States)

    Liu, Yangdan; Wang, Xin; Liu, Youping; Di, Xin

    2017-07-21

    The objective of this study was to develop a thermosensitive in situ gel based on solid dispersions (SDs) for rectal delivery of ibuprofen (IBU). Thermosensitive (poloxamer 407) and mucoadhesive (hydroxypropylmethyl cellulose E5 and sodium alginate) polymers were used to prepare the in situ gel and the sol-gel transition temperature (T sol-gel) and gel strength were optimized. The in vitro release performance and in vivo pharmacokinetic properties of the in situ gel after their rectal administration to rabbits were investigated. Compared with the solid suppository, the cumulative release of the IBU SDs loaded in situ gel was significantly increased. The in vivo pharmacokinetics indicated that in situ gel had a higher peak plasma concentration (C max) and area under the curve (AUC(0-∞)) in plasma than the solid suppositories. Histopathology results showed that the IBU in situ gel given at a dose of 15 mg/kg did not produce any irritation. In conclusion, this study suggested that the in situ gel could be an effective rectal formulation for IBU.

  20. Hydrocolloid-based nutraceutical delivery systems: Effect of counter-ions on the encapsulation and release.

    Science.gov (United States)

    Polowsky, Patrick J; Janaswamy, Srinivas

    2015-01-01

    Nutraceuticals provide health benefits, especially for the prevention and treatment of chronic diseases such as diabetes, obesity, cardiovascular disease and cancer. Their incorporation in food supplements, functional foods and medicinal foods is a major technological challenge due to lower water solubility, instability during processing and storage conditions. Carriers that can effectively overcome these predicaments and protect them during product development, consumption and delivery are in high demand. Toward this end, our research approach is to entrap nutraceuticals in the ordered networks of hydrocolloids. We have examined the effect cations in regulating the encapsulated amounts and release characteristics. Iota-carrageenan and eugenol have been chosen as models of hydrocolloid and nutraceutical, respectively, in the presence of Na and Ca ions. The results suggest that carrageenan maintains its network organization even after encapsulating the eugenol molecules. Increased eugenol amounts are found in the Na carrageenan complex compared to the Ca complex, and the release rate is faster from the former but it is more controlled from the latter. These differences highlight the vital role of cations on the encapsulation efficiency and release profiles of hydrocolloid-based nutraceutical carriers. The outcome offers an elegant opportunity for developing novel and value-added food systems employing low-in-cost, nontoxic and heavily consumed food grade hydrocolloids.

  1. A novel amphiphilic nano hydrogel using ketene based polyester with polyacrylamide for controlled drug delivery system.

    Science.gov (United States)

    Swarnalatha, S; Gopi, R; Ganesh Kumar, A; Selvi, P K; Sekaran, G

    2008-09-01

    A ketene based Low molecular weight polymer (LMKP) having ester functional group was prepared using glycine through surface initiated anionic polymerization. NMR, ATR-FTIR & SEC were used to characterize the LMKP. The LMKP and acrylamide (AAm) were co-polymerised in methyl ethyl ketone to yield semi-IPN nanohydrogels (NHG). Benzoyl peroxide (BPO) was used as an initiator and N,N-methylene bisacrylamide (MBA) as crosslinking agent. Formation of NHG was confirmed through frequency shift in LMKP and poly acrylamide (PAAm) in FTIR spectroscopy. Photon correlation spectroscopy reveals that the sizes of the NHG were in the range of 140-225 nm and Transmission Electron Micrograph (TEM) also confirms the nano dimension of NHG. Biocompatibility of the NHG was confirmed through the cytotoxicity analysis. The swelling and diffusion behaviour of NHG, prepared under various formulations, were evaluated. The swelling pattern of NHG was studied at different pH conditions. The drug delivery capacity of NHG was investigated using ciprofloxacin as a model drug. The drug release kinetics of NHG suggested their anomalous (non-fickian) behaviour.

  2. Development of a new controlled pesticide delivery system based on neem leaf powder

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Baljit, E-mail: baljitsinghhpu@yahoo.com [Department of Chemistry, Himachal Pradesh University, Shimla 171005 (India); Sharma, D.K.; Kumar, Ramesh; Gupta, Atul [Department of Chemistry, Himachal Pradesh University, Shimla 171005 (India)

    2010-05-15

    In order to minimize the agro-environmental pollution and health hazards caused by pesticides, in the present study, the neem leaf powder '(NLP)', a bio-pesticide, has been exploited to develop the pesticide delivery devices. The presence of neem in the formulations along with the pesticide may enhance the potential of these systems due to its inherent pesticidal activity. We have prepared the NLP and alginate based beads by using CaCl{sub 2} as crosslinker. To study the effect of composition of the beads on the release dynamics of fungicide (thiram), beads were prepared by varying the amount of NLP and crosslinker. The beads formed were characterized with Fourier transform infrared spectroscopy (FTIR), scanning electron micrograph (SEM), energy dispersion analysis by X-rays (EDAX), thermogravimetric analysis and swelling study. Formulation characteristics such as entrapment efficiency, bead size, percentage equilibrium swelling of the beads and diffusion mechanism for thiram release have been evaluated. Maximum (78.33 {+-} 2.89)% swelling has occurred in the beads prepared with 1.5% NLP, 2.5% alginate and 0.1 M crosslinker solution. In most of the formulations the values for the diffusion exponent 'n' have been obtained >1 and hence the release of fungicides occurred through Case II diffusion mechanism.

  3. Nanotechnology-based drug delivery systems for Alzheimer's disease management: Technical, industrial, and clinical challenges.

    Science.gov (United States)

    Wen, Ming Ming; El-Salamouni, Noha S; El-Refaie, Wessam M; Hazzah, Heba A; Ali, Mai M; Tosi, Giovanni; Farid, Ragwa M; Blanco-Prieto, Maria J; Billa, Nashiru; Hanafy, Amira S

    2017-01-10

    Alzheimer's disease (AD) is a neurodegenerative disease with high prevalence in the rapidly growing elderly population in the developing world. The currently FDA approved drugs for the management of symptomatology of AD are marketed mainly as conventional oral medications. Due to their gastrointestinal side effects and lack of brain targeting, these drugs and dosage regiments hinder patient compliance and lead to treatment discontinuation. Nanotechnology-based drug delivery systems (NTDDS) administered by different routes can be considered as promising tools to improve patient compliance and achieve better therapeutic outcomes. Despite extensive research, literature screening revealed that clinical activities involving NTDDS application in research for AD are lagging compared to NTDDS for other diseases such as cancers. The industrial perspectives, processability, and cost/benefit ratio of using NTDDS for AD treatment are usually overlooked. Moreover, active and passive immunization against AD are by far the mostly studied alternative AD therapies because conventional oral drug therapy is not yielding satisfactorily results. NTDDS of approved drugs appear promising to transform this research from 'paper to clinic' and raise hope for AD sufferers and their caretakers. This review summarizes the recent studies conducted on NTDDS for AD treatment, with a primary focus on the industrial perspectives and processability. Additionally, it highlights the ongoing clinical trials for AD management. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Ionically cross-linkable hyaluronate-based hydrogels for injectable cell delivery.

    Science.gov (United States)

    Park, Honghyun; Woo, Eun Kyung; Lee, Kuen Yong

    2014-12-28

    Although hyaluronate is an attractive biomaterial for many biomedical applications, hyaluronate hydrogels are generally formed using chemical cross-linking reagents that may cause unwanted side effects, including toxicity. We thus propose to design and prepare ionically cross-linkable hyaluronate compounds that can form gels in the presence of counter-ions. This study is based on the hypothesis that introduction of alginate to hyaluronate backbones (hyaluronate-g-alginate) could allow for gel formation in the presence of calcium ions. Here, we demonstrated ease of formation of cross-linked structures with calcium ions without additional chemical cross-linking reagents in hyaluronate-g-alginate (HGA) gels. The mechanical properties of HGA gels were regulated through changes in polymer composition and calcium concentration. We also confirmed that HGA gels could be useful in regenerating cartilage in a mouse model following subcutaneous injection into the dorsal region with primary chondrocytes. This finding was supported by histological and immunohistochemical analyses, glycosaminoglycan quantification and chondrogenic marker gene expression. This approach to the design and tailoring of ionically cross-linkable biomedical polymers may be broadly applicable to the development of biomaterials, especially in the drug delivery and tissue engineering fields. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Water based scale-up of CPO-27 synthesis for nitric oxide delivery.

    Science.gov (United States)

    Cattaneo, Damiano; Warrender, Stewart J; Duncan, Morven J; Castledine, Richard; Parkinson, Nigel; Haley, Ian; Morris, Russell E

    2016-01-14

    The applicability of water-based reflux and room temperature synthesis processes for the production of CPO-27 MOFs, suitable for NO delivery applications, is investigated. NO adsorption, storage and release performance of products obtained under reflux conditions are comparable to those of equivalent samples synthesised from traditional solvothermal methods at small scale. Products obtained from room temperature processes show lower NO release capability, although the quantities that are released are still more than adequate for biomedical applications. Results also reveal differences for the first time in NO uptake, storage and release depending on whether Zn, Ni or Mg is employed. The results indicate that while the crystallinity of CPO-27(Zn) and CPO-27(Mg) is not affected by moving to lower temperature methods, the crystallinity of CPO-27(Ni) is reduced. Particle morphology and size is also affected. The low temperature processes are successfully demonstrated at 20 L and 100 L scale and the main problems encountered during scale-up are outlined. The 100 L scale is in itself an appropriate production scale for some niche biomedical products. Indeed, results indicate that this synthesis approach is suitable for commercial production of MOFs for this application field. We also confirm that BET surface area from nitrogen adsorption at 77 K is not a good indicator for successful adsorption of NO.

  6. Long-term antibiotic delivery by chitosan-based composite coatings with bone regenerative potential

    Science.gov (United States)

    Ordikhani, F.; Simchi, A.

    2014-10-01

    Composite coatings with bone-bioactivity and drug-eluting capacity are considered as promising materials for titanium bone implants. In this work, drug-eluting chitosan-bioactive glass coatings were fabricated by a single-step electrophoretic deposition technique. Drug-loading and -releasing capacity of the composite coatings were carried out using the vancomycin antibiotic. Uniform coatings with a thickness of ∼55 μm containing 23.7 wt% bioactive glass particles and various amounts of the antibiotic (380-630 μg/cm2) were produced. The coatings were bioactive in terms of apatite-forming ability in simulated body fluid and showed favorable cell adhesion and growth. In vitro biological tests also indicated that the composite coatings had better cellular affinity than pristine chitosan coatings. The in vitro elution kinetics of the composite coating revealed an initial burst release of around 40% of the drug within the first elution step of 1 h and following by a continuous eluting over 4 weeks, revealing long-term drug-delivering potential. Antibacterial tests using survival assay against Gram-positive Staphylococcus aureus bacteria determined the effect of vancomycin release on reduction of infection risk. Almost no bacteria were survived on the coatings prepared from the EPD suspension containing ≥0.5 g/l vancomycin. The developed chitosan-based composite coatings with bone bioactivity and long-term drug-delivery ability may be potentially useful for metallic implants to reduce infection risk.

  7. Bioadhesive okra polymer based buccal patches as platform for controlled drug delivery.

    Science.gov (United States)

    Kaur, Gurpreet; Singh, Deepinder; Brar, Vivekjot

    2014-09-01

    In the present investigation, polysaccharide from the Okra fruits (Hibiscus esculentus) was extracted, characterized and explored for its mucoadhesive potential. Mucoadhesive films of okra polymer (OP) were prepared by solvent casting method based on 3(2) factorial design. For these studies, OP (2.0%, 2.5%, 3.0%, w/v) and glycerol (plasticizer) (0.25%, 0.50%, 0.75%, v/v) were taken as independent variables while tensile strength, mucoadhesive strength, contact angle, swelling index and residence time as dependent variables. The developed films were evaluated for their physicochemical, mechanical and electrical properties. The formulated films were found to be smooth, flexible, and displayed adequate mucoadhesive and tensile strength. Their near neutral pH and negative hemolytic studies indicated their non-irritability and biocompatible nature with biological tissues. The formulation comprising of 3% OP and 0.5% glycerol (F8) was found to exhibit optimum mechanical properties. Further, optimized film was loaded with zolmitriptan (model drug) to determine its drug release profiles. In vitro and ex vivo drug release studies demonstrated a controlled release of zolmitriptan over a period of 8h in simulated salivary fluid (SSF) pH 6.8, with the correlation coefficient values indicating its non-Fickian kinetics. Thus, OP can be used as a promising biomaterial for controlled drug delivery. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Rationale and Considerations for the Internet-Based Delivery of Parent-Child Interaction Therapy

    Science.gov (United States)

    Comer, Jonathan S.; Furr, Jami M.; Cooper-Vince, Christine; Madigan, Ryan J.; Chow, Candice; Chan, Priscilla; Idrobo, Fabio; Chase, Rhea M.; McNeil, Cheryl B.; Eyberg, Sheila M.

    2014-01-01

    Given the enormous individual, familial, and societal costs associated with early disruptive behavior disorders, transformative efforts are needed to develop innovative options for overcoming traditional barriers to effective care and for broadening the availability of supported interventions. This paper presents the rationale and key considerations for a promising innovation in the treatment of early-onset disruptive behavior disorders—that is, the development of an Internet-based format for the delivery of Parent-Child Interaction Therapy (PCIT) directly to families in their own homes. Specifically, we consider traditional barriers to effective care, and discuss how technological innovations can overcome problems of treatment availability, accessibility, and acceptability. We then detail our current Internet-delivered PCIT treatment program (I-PCIT), which is currently being evaluated across multiple randomized clinical trials relative to waitlist comparison, and to traditional in-office PCIT. Embedded video clips of children treated with I-PCIT are used to illustrate novel aspects of the treatment. PMID:26120268

  9. Formation of thermally reversible optically transparent emulsion-based delivery systems using spontaneous emulsification.

    Science.gov (United States)

    Saberi, Amir Hossein; Fang, Yuan; McClements, David Julian

    2015-12-28

    Transparent emulsion-based delivery systems suitable for encapsulating lipophilic bioactive agents can be fabricated using low-energy spontaneous emulsification methods. These emulsions are typically fabricated from non-ionic surfactants whose hydrophilic head groups are susceptible to dehydration upon heating. This phenomenon may promote emulsion instability due to enhanced droplet coalescence at elevated temperatures. Conversely, the same phenomenon can be used to fabricate optically transparent emulsions through the phase inversion temperature (PIT) method. The purpose of the current study was to examine the influence of oil phase composition and surfactant-to-oil ratio on the thermal behavior of surfactant-oil-water systems containing limonene, medium chain triglycerides (MCT), and Tween 60. Various types of thermal behavior (turbidity versus temperature profiles) were exhibited by these systems depending on their initial composition. For certain compositions, thermoreversible emulsions could be formed that were opaque at high temperatures but transparent at ambient temperatures. These systems may be particularly suitable for the encapsulation of bioactive agents in applications where optical clarity is important.

  10. Engineering of Δ9-tetrahydrocannabinol delivery systems based on surface modified-PLGA nanoplatforms.

    Science.gov (United States)

    Martín-Banderas, Lucía; Muñoz-Rubio, Inmaculada; Álvarez-Fuentes, Josefa; Durán-Lobato, Matilde; Arias, José L; Holgado, Ma Ángeles; Fernández-Arévalo, Mercedes

    2014-11-01

    The objective of this work is to develop a nanoplatform that can potentiate the oral administration of Δ9-tetrahidrocannabinol, a highly lipophilic active agent with very promising antiproliferative and antiemetic activities. To that aim, colloidal carriers based on the biodegradable and biocompatible poly(D,L-lactide-co-glycolide) were investigated. Such delivery systems were prepared by nanoprecipitation, and nanoparticle engineering further involved surface modification with a poly(ethylene glycol), chitosan, or poly(ethylene glycol)-chitosan shells to assure the greatest uptake by intestinal cells and to minimize protein adsorption. Characterization of the nanoplatforms included particle geometry (size and shape), electrophoretic properties (surface charge). Δ9-tetrahydrocannabinol vehiculization capabilities (loading and release), blood compatibility, and cellular uptake and cytotoxicity. Results were satisfactorily used to define the optimum engineering conditions to formulate surface modified nanoparticles for the efficient oral administration of Δ9-tetrahydrocannabinol. To the best of our knowledge, this is the first time that biocompatible polymeric nanoparticles have been formulated for Δ9-tetrahydrocannabinoldelivery. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Water soluble biocompatible vesicles based on polysaccharides and oligosaccharides inclusion complexes for carotenoid delivery.

    Science.gov (United States)

    Polyakov, Nikolay E; Kispert, Lowell D

    2015-09-05

    Since carotenoids are highly hydrophobic, air- and light-sensitive hydrocarbon compounds, developing methods for increasing their bioavailability and stability towards irradiation and reactive oxygen species is an important goal. Application of inclusion complexes of "host-guest" type with polysaccharides and oligosaccharides such as arabinogalactan, cyclodextrins and glycyrrhizin minimizes the disadvantages of carotenoids when these compounds are used in food processing (colors and antioxidant capacity) as well as for production of therapeutic formulations. Cyclodextrin complexes which have been used demonstrated enhanced storage stability but suffered from poor solubility. Polysaccharide and oligosaccharide based inclusion complexes play an important role in pharmacology by providing increased solubility and stability of lipophilic drugs. In addition they are used as drug delivery systems to increase absorption rate and bioavailability of the drugs. In this review we summarize the existing data on preparation methods, analysis, and chemical reactivity of carotenoids in inclusion complexes with cyclodextrin, arabinogalactan and glycyrrhizin. It was demonstrated that incorporation of carotenoids into the "host" macromolecule results in significant changes in their physical and chemical properties. In particular, polysaccharide complexes show enhanced photostability of carotenoids in water solutions. A significant decrease in the reactivity towards metal ions and reactive oxygen species in solution was also detected. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Faraday instability-based micro droplet ejection for inhalation drug delivery

    Science.gov (United States)

    Tsai, C.S.; Mao, R.W.; Lin, S.K.; Zhu, Y.; Tsai, S.C.

    2014-01-01

    We report here the technology and the underlying science of a new device for inhalation (pulmonary) drug delivery which is capable of fulfilling needs unmet by current commercial devices. The core of the new device is a centimeter-size clog-free silicon-based ultrasonic nozzle with multiple Fourier horns in resonance at megahertz (MHz) frequency. The dramatic resonance effect among the multiple horns and high growth rate of the MHz Faraday waves excited on a medicinal liquid layer together facilitate ejection of monodisperse droplets of desirable size range (2–5 µm) at low electrical drive power (<1.0 W). The small nozzle requiring low drive power has enabled realization of a pocket-size (8.6 × 5.6 × 1.5 cm3) ultrasonic nebulizer. A variety of common pulmonary drugs have been nebulized using the pocket-size unit with desirable aerosol sizes and output rate. These results clearly provide proof-of-principle for the new device and confirm its potential for commercialization. PMID:25045720

  13. Multidrug Delivery Systems Based on Human Serum Albumin for Combination Therapy with Three Anticancer Agents.

    Science.gov (United States)

    Qi, Jinxu; Zhang, Yao; Gou, Yi; Lee, Philbert; Wang, Jun; Chen, Shifang; Zhou, Zuping; Wu, Xiaoyang; Yang, Feng; Liang, Hong

    2016-09-06

    When administering several anticancer drugs within a single carrier, it is important to regulate their spatial distribution so as to avoid possible mutual interference and to thus enhance the drugs' selectivity and efficiency. To achieve this, we proposed to develop human serum albumin (HSA)-based multidrug delivery systems for combination anticancer therapy. We used three anticancer agents (an organic drug [5-fluorouracil, or 5FU], a metallic agent [2-benzoylpyridine thiosemicarbazide copper II, or BpT], and a gene agent [AS1411]) to treat liver cancer and confirm our hypothesis. The structure of the HSA-palmitic acid (PA)-5FU-BpT complex revealed that 5FU and BpT, respectively, bind to the IB and IIA subdomains of HSA. Our MALDI-TOF-MS spectral data show that one AS1411 molecule is conjugated to Cys-34 of the HSA-5FU-BpT complex via a linker. Compared with unregulated three-drug combination therapy, the HSA-5FU-BpT-AS1411 complex enhances cytotoxicity in Bel-7402 cells approximately 7-fold in vitro; however, in normal cells it does not raise cytotoxicity levels. Importantly, our in vivo results demonstrate that the HSA-5FU-BpT-AS1411 complex is superior to the unregulated three-drug combination in enhancing targeting ability, inhibiting liver tumor growth, and causing fewer side effects.

  14. Reputation-Based Colluder Detection Schemes for Peer-to-Peer Content Delivery Networks

    National Research Council Canada - National Science Library

    ABDULLAH, Ervianto; FUJITA, Satoshi

    2013-01-01

    Recently Peer-to-Peer Content Delivery Networks (P2P CDNs) have attracted considerable attention as a cost-effective way to disseminate digital contents to paid users in a scalable and dependable manner...

  15. Self-assembled peptide-based nanostructures: Smart nanomaterials toward targeted drug delivery.

    Science.gov (United States)

    Habibi, Neda; Kamaly, Nazila; Memic, Adnan; Shafiee, Hadi

    2016-02-01

    Self-assembly of peptides can yield an array of well-defined nanostructures that are highly attractive nanomaterials for many biomedical applications such as drug delivery. Some of the advantages of self-assembled peptide nanostructures over other delivery platforms include their chemical diversity, biocompatibility, high loading capacity for both hydrophobic and hydrophilic drugs, and their ability to target molecular recognition sites. Furthermore, these self-assembled nanostructures could be designed with novel peptide motifs, making them stimuli-responsive and achieving triggered drug delivery at disease sites. The goal of this work is to present a comprehensive review of the most recent studies on self-assembled peptides with a focus on their "smart" activity for formation of targeted and responsive drug-delivery carriers.

  16. The use of liquid self-emulsifying drug delivery systems based on ...

    African Journals Online (AJOL)

    emulsifying drug delivery systems (LSEDDS) containing griseofulvin. The LSEDDS were evaluated using the following parameters: phase separation, globule size, viscosity, solubility of griseofulvin and partition coefficient. The release profile of ...

  17. Facility-based delivery and maternal and early neonatal mortality in ...

    African Journals Online (AJOL)

    AJRH Managing Editor

    including such things as cost, distance to facilities, perceived need ..... system, allowing traditional and contemporary ..... women's choice of place of delivery in rural Malawi-- .... incentives: early results from a quasi-experimental study. BMC Int ...