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Full-Text Articles in Dentistry
Enhanced Gene Delivery In Vitro And In Vivo By Improved Transferrin-Lipoplexes, Conchita Tros De Ilarduya, M. A. Arangoa, M. J. Moreno-Aliaga, Nejat Düzgüneş
Enhanced Gene Delivery In Vitro And In Vivo By Improved Transferrin-Lipoplexes, Conchita Tros De Ilarduya, M. A. Arangoa, M. J. Moreno-Aliaga, Nejat Düzgüneş
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Cationic liposomes and the complexes they form with DNA (lipoplexes) constitute the most promising alternative to the use of viral vectors for gene therapy. One of the limitations to their application in vivo, however, is the inhibition of gene delivery by serum. In a previous study, we demonstrated that transferrin (Tf)-lipoplexes were superior to plain lipoplexes in transfecting HeLa cells in the presence of high concentrations of serum. With the goal of obtaining efficient gene expression in vivo, we evaluated the efficacy of Tf-lipoplexes (containing DOTAP and cholesterol) in transfecting primary hepatocytes and adipocytes in the presence of high serum …
Efficient Gene Transfer By Transferrin Lipoplexes In The Presence Of Serum, Conchita Tros De Ilarduya, Nejat Düzgüneş
Efficient Gene Transfer By Transferrin Lipoplexes In The Presence Of Serum, Conchita Tros De Ilarduya, Nejat Düzgüneş
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Cationic lipids are being used increasingly as reagents for gene delivery both in vitro and in vivo. One of the limitations to the application of cationic lipid-DNA complexes (lipoplexes) in vivo is the inhibition of gene delivery by serum. In this study, we have shown that transferrin (Tf)-lipoplexes, which had transferrin adsorbed at their surface via electrostatic interactions, are much more effective than plain lipoplexes in transfecting cells in the presence of relatively high concentrations (up to 60%) of fetal bovine serum (FBS). Serum even enhanced transfection by Tf-lipoplexes composed of 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP)/dioleoylphosphatidylethanolamine (DOPE)/pCMVLacZ at high lipid/DNA (+/-) charge …
Human Serum Albumin Enhances Dna Transfection By Lipoplexes And Confers Resistance To Inhibition By Serum, Sérgio Simões, Vladimir Slepushkin, Pedro Pires, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
Human Serum Albumin Enhances Dna Transfection By Lipoplexes And Confers Resistance To Inhibition By Serum, Sérgio Simões, Vladimir Slepushkin, Pedro Pires, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
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Cationic liposome-DNA complexes ('lipoplexes') are used as gene delivery vehicles and may overcome some of the limitations of viral vectors for gene therapy applications. The interaction of highly positively charged lipoplexes with biological macromolecules in blood and tissues is one of the drawbacks of this system. We examined whether coating cationic liposomes with human serum albumin (HSA) could generate complexes that maintained transfection activity. The association of HSA with liposomes composed of 1,2-dioleoyl-3-(trimethylammonium) propane and dioleoylphosphatidylethanolamine, and subsequent complexation with the plasmid pCMVluc greatly increased luciferase expression in epithelial and lymphocytic cell lines above that obtained with plain lipoplexes. The …
Interaction Of Cationic Liposomes And Their Dna Complexes With Monocytic Leukemia Cells, Pedro Pires, Sérgio Simões, Shlomo Nir, Rogério Gaspar, Nejat Düzgüneş, Maria C. Pedroso De Lima
Interaction Of Cationic Liposomes And Their Dna Complexes With Monocytic Leukemia Cells, Pedro Pires, Sérgio Simões, Shlomo Nir, Rogério Gaspar, Nejat Düzgüneş, Maria C. Pedroso De Lima
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Cationic liposomes complexed with DNA have been used extensively as non-viral vectors for the intracellular delivery of reporter or therapeutic genes in culture and in vivo. We examined the relationship between the characteristics of the lipoplexes, their mode of interaction with monocytic THP-1 cells and their ability to transfect these cells. We determined the size and ζ potential of cationic liposomes (composed of 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP) and its mixtures with neutral lipids), and lipoplexes at different (+/-) charge ratios. As the (+/-) charge ratio of the lipoplexes decreased to (1/1), a significant reduction in ζ potential and an increase in …
Transfection Of Human Macrophages By Lipoplexes Via The Combined Use Of Transferrin And Ph-Sensitive Peptides, Sérgio Simões, Vladimir Slepushkin, Elizabeth Pretzer, Paul Dazin, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
Transfection Of Human Macrophages By Lipoplexes Via The Combined Use Of Transferrin And Ph-Sensitive Peptides, Sérgio Simões, Vladimir Slepushkin, Elizabeth Pretzer, Paul Dazin, Rogério Gaspar, Maria C. Pedroso De Lima, Nejat Düzgüneş
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The crucial function of macrophages in a variety of biological processes and pathologies render these cells important targets for gene therapeutic interventions. Commonly used synthetic gene delivery vectors have not been successful in transfecting these non-dividing cells. A combination strategy involving cationic liposomes to condense and carry DNA, transferrin to facilitate cellular uptake, and the pH-sensitive peptide GALA to promote endosome destabilization, resulted in significant expression of a luciferase gene. Transfection of macrophages was dependent on the degree of differentiation of the cells. The quaternary complexes of cationic liposomes, DNA, transferrin, and GALA exhibited a net negative charge, which may …
Delivery Of An Anti-Hiv-1 Ribozyme Into Hiv-Infected Cells Via Cationic Liposomes, Krystyna Konopka, John J. Rossi, Piotr Swiderski, Vladimir A. Slepushkin, Nejat Düzgüneş
Delivery Of An Anti-Hiv-1 Ribozyme Into Hiv-Infected Cells Via Cationic Liposomes, Krystyna Konopka, John J. Rossi, Piotr Swiderski, Vladimir A. Slepushkin, Nejat Düzgüneş
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Cationic liposome-mediated intracellular delivery of a fluorescein-labeled chimeric DNA-RNA ribozyme targeted to the HIV-1 5' LTR was investigated, using THP-1, THP-1/HIV-1(IIIB) or HeLa/LAV cells. Different fluorescence patterns were observed when the cells were exposed to Lipofectamine, Lipofectin or DMRIE:DOPE (1:1) complexed to the ribozyme. With Lipofectamine intense cell-associated fluorescence was found. Incubation with Lipofectin resulted in less intense diffuse fluorescence, while with DMRIE an intense but sporadic fluorescence was observed. Differentiated THP-1/HIV-1(IIIB) cells were more susceptible to killing by liposome-ribozyme complexes than THP-1 cells. Under non-cytotoxic conditions (a 4-h treatment) complexes of 5, 10 or 15 μM Lipofectin or DOTAP:DOPE …
Human Immunodeficiency Virus Type-1 (Hiv-1) Infection Increases The Sensitivity Of Macrophages And Thp-1 Cells To Cytotoxicity By Cationic Liposomes, Krystyna Konopka, Elizabeth Pretzer, Philip L. Felgner, Nejat Düzgüneş
Human Immunodeficiency Virus Type-1 (Hiv-1) Infection Increases The Sensitivity Of Macrophages And Thp-1 Cells To Cytotoxicity By Cationic Liposomes, Krystyna Konopka, Elizabeth Pretzer, Philip L. Felgner, Nejat Düzgüneş
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Cationic liposomes may be valuable for the delivery of anti-sense oligonucleotides, ribozymes, and therapeutic genes into human immunodeficiency virus type 1 (HIV-1)-infected and uninfected cells. We evaluated the toxicity of three cationic liposomal preparations, Lipofectamine, Lipofectin, and 1,2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide (DMRIE) reagent, to HIV-infected and uninfected cells. Monocyte/macrophages were infected with HTV-1(BaL) and treated with liposomes in medium containing 20% fetal bovine serum (FBS) for 4 h or 24 h at 37°C. Uninfected monocytic THP-1 cells and chronically infected THP-1/HIV-1(IIIB) cells were treated with phorbol 12-myristate 13-acetate (PMA) and exposed to liposomes in the presence of 10% FBS. Toxicity was …