The transfection efficiency of DNA electroporation was compared with that of non-electroporation methods including, liposome-DNA complexes and integrin-liposome-DNA complexes in different. In general, electroporation can be optimized to control survival rate within 30-40 % and to maximize transfection efficiency (Zhou et al. Electroporation has been successfully used to administer HPV DNA vaccine to mice as well as rhesus macaques, which has prompted its use in an ongoing phase I clinical trial such as VGX-3100, a vaccine that includes plasmids targeting E6 and E7 proteins of both HPV subtypes 16 and 18, for treatment of patients with CIN 2 or 3. DNA formulation with certain types of polymers has been found to enhance electroporation efficiency and, in some cases, reduce treatment-related toxicity. Disadvantages include potential cell damage and the nonspecific transport of molecules into and out of the cell. We will talk about the advantages of electrical energy , this being a source of energy that can be achieved thanks to the movement of charges rich in negative and positive electrons. A wide range of pulse patterns have been used both in vitro and in vivo. The effects of electroporation buffer composition on cell - Nature Figue 2 shows several important EP-mediated DNA vaccines used in clinical trials. Matej Reberek, Damijan Miklavi Advantages and Disadvantages of Another approach involves employing a method capable of improving the intramuscularly delivery of protein-based vaccine led to the slow release of the protein. Gold nanoparticles devoid of DNA coating were not taken up by cells during electroporation. Typically, in vivo electroporation is performed by first injecting DNA to the target tissue followed by electric pulses, with varied voltage, pulse duration and number of cycles, from two applied electrodes [Al-Dosari and Gao, 2009, Hao et al., 2012]. The authors have shown that if the voltage of the pulses exceeds a voltage threshold at 75100 V (equivalent to the breakdown threshold of 810 lipid bilayers in the SC), microchannels or local transport regions are created through the breakdown sites of the SC [Hui, 2013]. Bleomycin electrochemotherapy has been successfully applied to treat melanomas, head and neck squamous cell carcinomas, Kaposis sarcomas, as well as lung, breast, kidney, and bladder cancers. For Research Use Only. However, further optimization of DNA vaccine delivery is needed for this vaccine modality to ultimately be efficacious in humans [Hallengrd et al., 2012]. By making research easy to access, and puts the academic needs of the researchers before the business interests of publishers. Different non-viral approaches have been proposed for drug and gene delivery such as physical and chemical methods. The use of electroporation pulses enhancing the skin permeability to deliver anti-viral drugs is in the early stages of development. Our team is growing all the time, so were always on the lookout for smart people who want to help us reshape the world of scientific publishing. PDF Advantages and Disadvantages of Different Concepts of Electroporation Since every tissue is specific and has its own characteristics, there are no generally accepted optimal conditions of electroporation that are suitable for effective transfection. Not for use in diagnostic procedures. The effect of electroporation on DNA vaccine potency and gene delivery was studied using skin as a target tissue in larger animal species such as pig, macaque and sheep. The enhancement of luciferase activity was observed in both small and large animal species. Studies using electroporation were performed in vivo; however electroporation is sometimes harmful to differentiated adult cells [Anwer, 2011; Wang et al., 2012]. PMID: 25981214 DOI: 10.1016/j.diii.2015.04.007 Abstract Several ablation techniques are currently available. In vivo plasmid electroporation has also been used as either a primary or booster vaccination strategy that enhances cell-mediated immune responses. Using this approach, highly efficient gene transfer has already been achieved in muscle and liver as well as in tumors [Tamura and Sakata, 2003]. For instance, a number of studies have demonstrated long-term, complete tumor regression, using delivery of plasmids encoding IL-12 or IFN- as a single agent in melanoma and squamous cell carcinoma (SCC) [Heller and Heller, 2006]. The combination of iontophoresis and electroporation was found to be effective in delivering siRNA but not plasmid DNA into the corneal epithelium [Hao et al., 2010]. These techniques show a potential for drug and gene delivery. In addition, several DNA formulations have been described for in vivo gene electroporation. A consistent finding is that lipo-or amphiphilic drugs traverse the cell membrane without electroporation, while an enhancement in cytotoxicity is found with drugs that, under normal circumstances, do not pass the cell membrane easily. Open Access is an initiative that aims to make scientific research freely available to all. Most of the studies using electroporation have involved local delivery into the target organ but a few have studied local electroporation following systemic (intravenous) delivery of the plasmid. Personally, I prefer chemical transformation. Electroporation is a transient phenomenon that increases the per- meability of cell plasma . Publishing on IntechOpen allows authors to earn citations and find new collaborators, meaning more people see your work not only from your own field of study, but from other related fields too. Electroporation - Advantages and Drawbacks for Delivery of Drug, Gene The in vitro and in vivo studies using electroporation have been further described as following: In vitro electroporation: Electroporation can be used to transfer a range of genetic materials into cells including DNA, RNA and oligonucleotides. In vivo electroporation as compared to other gene transfer methods, such as viral vectors, has several advantages: a) various types of DNA constructs (or RNAi vectors) are readily introduced to the cells without limitation of DNA size; b) more than two different DNA constructs can be introduced into the same cells [Matsuda and Cepko, 2007]. Assay cells for gene expression or silencing. This is an important consideration for ex vivo gene delivery especially to hematopoietic cells. The 1 mL electroporation chamber enables efficient process development and scales directly to commercial manufacturing using the 525 mL cartridge. An equal variety of electrodes have been developed for in vivo use, based on the nature of the tissue being treated [Wells, 2010]. An electrical pulse at an optimized voltage and only lasting a few microseconds to a millisecond is discharged through the cell suspension. Many small-molecule drugs have been successfully delivered through the skin by electroporation. This review examines the advantages and disadvantages of physical non-viral methods (i.e., microinjection, ballistic gene delivery, electroporation, sonoporation, laser irradiation, magnetofection, and electric field-induced molecular vibration), with particular attention given to electroporation because of its versatility, with further special . The researchers used the electric pulses to increase transdermal transport of D-aminolevulinic acid (ALA), a protoporphyrin IX (PpIX)-precursor for the photodynamic therapy of superficial skin cancer and cutaneous metastases of internal malignancies. Electric shocks are used as a mechanism for introducing new DNA into a host cell by creating new pores in the plasma membrane of the host cell. It was suggested that hydrophobic sections of poloxamer 188 molecules are incorporated into the edges of pores and that their hydrophilic parts act as brushy pore structures. Apart from the convenience and non-invasiveness, the skin also provides a reservoir that sustains delivery over a period of days [Hui, 2013]. However, optimization of pulse and field strength parameters is still required to balance electroporation efficiency and cell viability. Milder electroporation conditions, although less toxic, are transfectionally inefficient. The electroporation delivery was found to be superior to all other test methods. Using two different beta-blockers, the researchers showed that lipophilicity and positive charges affected the electrotransport of drugs [Escobar-Chvez et al., 2009]. Application of short electrical pulses can be used to enhance gene delivery and DNA vaccination in large animals led to improved cellular and humoral immune responses. The presence of BCD and HPCD enhanced the total transport of the permeants piroxicam and carboxyfluorescein (CF), respectively, from both permeant solutions and suspensions. At the Institut Gustave-Roussy, France, the fist clinical trial of ECT with bleomycin in eight patients with recurrent or progressive head and neck squamous cell carcinoma was published in 1991. Electroporation is a very popular and highly efficient method of transfection. Advantages and Disadvantages of Different Concepts of Electroporation Pulse Generation. Geraci F. Extracellular membrane vesicles as a mechanism of cell-to-cell communication: advantages and disadvantages. Electroporation is sensitive to salt - you can lose precious samples if excess salt is carried over into the cuvette. This variation in tumor response could be due to differences in the state of tumor necrosis, tumor conductivity, or matrix complexity between the different tumors [Anwer, 2008]. The success of in vitro delivery by electroporation has led to the development of in vivo applications [Takei et al., 2008]. An electroporation driven plasmid DNA vaccination strategy was studied in animal models for treatment of prostate cancer. The proprietary tip technology of Neon NxT Electroporation System enables scientists to reduce sample transfer loss while simplifying their workflow. Irreversible Electroporation. Hence, the use of targeted ligands is an attractive approach to improve target specificity of electroporation, but its in vivo application has not been fully established. Indeed, the effect of calcium electroporation is independent of calcium compound [Frandsen et al., 2014]. Electroporation-based immunization (especially, EP-mediated DNA vaccine) has been effective in a number of species including mice, rats, rabbits, non-human primates, pigs and sheep. In vivo use of electroporation is done by injecting naked DNA followed by electric pulses from electrodes that are located in situ in the target tissues. Importantly, these antibody responses were sustained for 25 weeks after vaccination [van Drunen Littel-van den Hurk et al., 2008]. Calcium electroporation offers several advantages over standard treatment options: calcium is inexpensive and may readily be applied without special precautions mentioned about cytostatic drugs. Steve Haltiwanger, in Electroporation-Based Therapies for Cancer, 2014. The thresholds for different cells in a heterogeneous tissue would thus vary [Hui, 2008]. 1. Electroporation has proven useful both in vitro, in vivo and in patients, where drug delivery to malignant tumors has been performed. [5] This can cause problems in studies where gene expression has to be controlled to ensure accurate and precise results. Non-viral vectors are safe in human body and easy for use. We are a community of more than 103,000 authors and editors from 3,291 institutions spanning 160 countries, including Nobel Prize winners and some of the worlds most-cited researchers. Electroporation of IL-5 plasmid DNA into mouse tibialis muscle produced 20 ng IL-5/mL while the non-electroporated delivery generated only 0.2 ng IL-5/mL in the blood. The further improvements of electrodes including shape or arrangement of electrodes and electric conditions, by which more efficient and reliable gene transfer is achieved, are important especially in clinical trials. How long can gene expression be maintained after electrotransfer? Plasmid DNA vaccination using skin electroporation (EP) is a promising method able to elicit robust humoral and CD8+T-cell immune responses while limiting invasiveness of delivery [Daemi et al., 2012]. The process of electroporation is fairly simplistic. The technical feasibility of in vivo DNA targeting by electroporation has not been fully established. Apply electrical pulse to cells in the presence of specialized buffer and nucleic acids. The results of the different regimens are comparable, but there may be more necrosis with the higher doses and a better chance to conserve normal tissue with the lower doses [Gehl, 2008]. These pulses generated transient pores in cell membranes followed by intracellular electrophoretic DNA movement. The tumor cell line treated with electroporation showed efficient drug delivery suggesting further cell death. The reports showed an enhanced transport of human luteinizing hormone releasing hormone through heat-stripped human epidermis by electroporation. Frontiers | Engineering NK Cells for CAR TherapyRecent Advances in The magnitude of liposome enhancement was dependent on the degree of lipid saturation but independent of polar head group [Anwer, 2008]. Optimization of the approach indicated that a four-dose regimen provided highest tumor protection. It was suggested that hydrophobic sections of poloxamer 188 molecules are incorporated into the edges of pores and their hydrophilic parts act as brushy pore structures. Engineering exosomes as refined biological nanoplatforms for drug Needle-free non-adjuvant skin immunization by electroporation has been reported [Hui, 2008]. Larger molecules, including heparin, polylysine, antisense polynucleotides, lactalbumin, and IgG, have been delivered by transdermal electroporation with proper enhancers. Brief introduction to this section that descibes Open Access especially from an IntechOpen perspective, Want to get in touch? An electroporation method was established to involve a constant voltage and plate and fork type electrodes and use it for in vivo delivery of siRNA. Electroporation was introduced in the 1960s and comprises the application of controlled electric fields to facilitate cell permeabilization. As for all transfection methods, electroporation has its advantages and disadvantages. Contact our London head office or media team here. There have also been limited reports of successful protein delivery by this mechanism [Sharei et al., 2013]. How? Its based on principles of collaboration, unobstructed discovery, and, most importantly, scientific progression. Electroporation - Davidson The initial study of in vivo EP was the delivery of chemotherapeutic agents to solid tumors. Figure 1. Non-viral vectors are attractive tools in gene therapy and vaccine delivery [Draghia-Akli et al., 2005]. Comparison of chemical transformation and electroporation. A number of approaches for enhancing the potency of DNA vaccines have developed over the past few years such as: a) Optimization of DNA constructs; b) Development of new DNA manufacturing processes and formulations; c) Augmentation of immune responses with novel encoded molecular adjuvants; and d) Improvement of in vivo DNA delivery strategies including electroporation [Sardesai and Weiner, 2011]. A higher interaction of positively charged lipid-DNA complexes with negatively charged cell surfaces could be one of the underlying mechanisms in the lipid enhancement of the electroporation. Lipofection - an overview | ScienceDirect Topics The type of a nucleic acid and the type of the transfected cell generally affect the efficiency of electroporation [Stroh et al., 2010]. This disturbs the phospholipid bilayer of the membrane and results in the formation of temporary pores. For example, local electroporation targeted plasmid delivery to the liver was effective for liver, kidney and spleen but was not successful for skeletal muscle or skin [Wells, 2010]. Notably, a long interval (20 days) of electroporation was enough to obtain a satisfactory effect. 5 Princes Gate Court, The purpose of this study was to investigate the feasibility of using irreversible electroporation as a liver-directed ablation technique for the treatment of hepatocellular carcinoma (HCC) in the N1-S1 rodent model. Much intensive research has gone into the development of safe and efficient methods for the delivery of therapeutic genes [Tamura and Sakata, 2003]. Physical Non-Viral Gene Delivery Methods for Tissue Engineering - Springer Electroporation is temporary destabilization of the cell membrane targeted tissue by insertion of a pair of electrodes into it so that DNA molecules in the surrounding . Recently, peptides and mini-gene vaccines are of particular interest since several epitopes of tumor-associated antigens have been employed as therapeutic and prophylactic cancer vaccines. A number of studies have demonstrated the feasibility of targeted delivery of oligonucleotides, small interfering RNA (siRNA), plasmid DNA, and viral vectors to the corneal cells in vivo, specifically stromal keratocytes and corneal epithelial cells, via intrastromal injection, iontophoresis, electroporation, and gene gun. In vivo electroporation depends on electric pulses to drive gene transfer. Although the principle of electroporation is applicable to all cell types, its efficiency depends on the electrical properties of the cells. The system was designed for electroporation of mammalian cells, but some customers have found it to be successful for other cell types such as insect cell cultures and parasites. Electroporation is a widely recognized method of gene delivery into mammalian tissues. The advantages of this electroporation device are: The design of the electroporation chamber distributes the current equally among the cells and maintains a stable pH throughout the chamber; these key benefits increase cell viability dramatically. Advantages and Disadvantages of Different Concepts of Electroporation Pulse Generation M. Reberek, . Furthermore, the electrode and patch design is an important issue to reduce the discomfort of the electrical treatment in humans [Escobar-Chvez et al., 2009]. Altogether, the use of suitable electric pulses could trigger changes in the cytoskeleton organization and cell adhesiveness, led to the enhancement of anti-tumor effects [Pehlivanova et al., 2012]. This method was used to examine whether introduction and expression of PPAR gene could differentiate skeletal muscle satellite cells to adipocytes in vivo [Bonamassa and Liu, 2010]. Furthermore, complete responses have been observed in a fibrosarcoma model after delivery of a plasmid encoding GM-CSF and B7.1 [Heller and Heller, 2006].
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