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Laurent J. Feldman - One of the best experts on this subject based on the ideXlab platform.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:1 - ArticleIn the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA.
Journal of biomedical materials research. Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethylamine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc.
Daniel Scherman - One of the best experts on this subject based on the ideXlab platform.
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Intramuscular Plasmid DNA electrotransfer: Biodistribution and degradation
Biochimica et biophysica acta, 2004Co-Authors: Michel Bureau, Johanne Seguin, S Naimi, R Torero Ibad, C Georger, E Arnould, L Maton, F Blanche, P Delaere, Daniel SchermanAbstract:Abstract We have studied radiolabelled Plasmid DNA biodistribution and degradation in the muscle at different times after injection, with or without electrotransfer using previously defined conditions. Radiolabelled Plasmid progressively left the muscle and was degraded as soon as 5 min after Plasmid injection, with or without electrotransfer. Autoradiography showed that the major part of injected radioactivity was detected in the interfibrilar space of a large proportion of the muscle. Large zones of accumulation of radioactivity, which seems to be contained in some fibres (more than 20 μm), were identified as soon as 5 min after electrotransfer. Such structures were never observed on slices of non-electrotransferred muscles. However, these structures were not frequent and probably lesional. The surprising fact is that despite the amount of intact Plasmid having been greatly reduced between 5 min and 3 h after injection, the level of transfection remains unchanged whether electric pulses were delivered 20 s or 3 h after injection. Such a behavior was similarly observed when injecting 0.3, 3 or 30 μg of Plasmid DNA. Moreover, the transfection level was correlated to the amount of Plasmid DNA injected. These results suggest that as soon as it is injected, Plasmid DNA is proportionally partitioned between at least two compartments. While a major part of Plasmid DNA is rapidly cleared and degraded, the electrotransferable pool of Plasmid DNA represents a very small part of the amount injected and belongs to another compartment where it is protected from endogenous DNAses.
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1038. Intramuscular Plasmid DNA Electrotransfer. Biodistribution, Degradation and Stability of the Transfectable Pool of Plasmid DNA
Molecular Therapy, 2004Co-Authors: Michel-francis Bureau, Naimi Sadoui, Johanne Seguin, Daniel SchermanAbstract:In the present work, we used Plasmid DNA radiolabelled with 3H by methylation (10) to study radiolabelled Plasmid DNA biodistribution and degradation in the muscle at different times after injection, with or without electrotransfer using previously defined conditions. Radiolabelled Plasmid progressively left the muscle and was degraded as soon as 5 minutes after Plasmid injection, with or without electrotransfer. Autoradiography showed that the major part of injected radioactivity was detected in the interfibrilar space of a large proportion of the muscle. Large zones of accumulation of radioactivity which seems to be into some fibres (more than 20 μm) were identified as soon as 5 minutes after electrotransfer. Such structures were never observed on slices of non-electrotransfered muscles. However, these structures were not frequent and probably lesional. The surprising fact is that despite the amount of intact Plasmid having been greatly reduced between 5 minutes and 3 hours after injection, the level of transfection remains unchanged whether electric pulses were delivered 20 sec, 1 hour, 2 hours or 3 hours after injection. Such a behavior was similarly observed when injecting 0.3, 3 or 30 μg of Plasmid DNA. Moreover, the transfection level was correlated to the amount of Plasmid DNA injected. These results suggest that as soon as it is injected, Plasmid DNA is proportionally partitioned between at least two compartments. While a major part of Plasmid DNA is rapidly cleared and degraded, the electrotransferable pool of Plasmid DNA represents a very small part of the amount injected and belongs to another compartment where it is protected from endogenous DNAses.
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Intracellular fate and nuclear targeting of Plasmid DNA.
Cell biology and toxicology, 1999Co-Authors: Carole Neves, Daniel Scherman, Virginie Escriou, Gerardo Byk, Pierre WilsAbstract:One of the major steps limiting nonviral gene transfer efficiency is the entry of Plasmid DNA from the cytoplasm into the nucleus of the transfected cells. The nuclear localization signal (NLS) of the SV40 large T antigen is known to efficiently induce nuclear targeting of proteins. We have developed two chemical strategies for covalent coupling of NLS peptides to Plasmid DNA. One method involves a site-specific labeling of Plasmid DNA by formation of a triple helix with an oligonucleotide–NLS peptide conjugate. After such modification with one NLS peptide per Plasmid molecule, Plasmid DNA remained fully active in cationic lipid-mediated transfection. In the other method, we randomly coupled 5–115 p-azidotetrafluorobenzyllissamine–NLS peptide molecules per Plasmid DNA by photoactivation. Oligonucleotide–NLS and Plasmid–lissamine–NLS conjugates interacted specifically with the NLS-receptor importin α. Plasmid–lissamine–NLS conjugates were not detected in the nucleus, after cytoplasmic microinjection. Plasmids did not diffuse from the site of injection and Plasmid–lissamine–NLS conjugates appeared to be progressively degraded in the cytoplasm. The process of Plasmid DNA sequestration/degradation stressed in this study might be as important in limiting the efficiency of nonviral gene transfer as the generally recognized entry step of Plasmid DNA from the cytoplasm into the nucleus
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efficient purification of Plasmid DNA for gene transfer using triple helix affinity chromatography
Gene Therapy, 1997Co-Authors: Pierre Wils, Virginie Escriou, A Warnery, F Lacroix, Delphine Lagneaux, M Ollivier, Joel Crouzet, Jeanfrancois Mayaux, Daniel SchermanAbstract:Plasmid DNA used for nonviral therapeutic gene transfer or nucleic acid vaccination has to be highly purified, devoid of contaminating components such as bacterial proteins, endotoxins, or bacterial chromosomal DNA. We have developed a new affinity chromatography technique for Plasmid DNA purification: triple-helix affinity chromatography (THAC). This technique is based on the sequence-specific interaction of an oligonucleotide forming a triple-helix with Plasmid DNA. The oligonucleotide was covalently linked to a chromatographic matrix, thus providing a reusable affinity support. By inserting a suitable homopurine sequence in the Plasmid DNA, it is possible to obtain a triple-helix interaction that will only be stable at mild acidic pH and that will dissociate in alkaline conditions. A crude lysate from a recombinant E. coli, or a pre-purified Plasmid DNA, is thus applied at acidic pH on to a THAC column. After extensive washing of the column, purified Plasmid DNA is eluted using an alkaline buffer. The binding conditions of the Plasmid DNA on to the column have been optimized, as well as the hybridization sequence and the linker group between the matrix and the third strand oligonucleotide. The THAC technique makes it possible to purify in one step supercoiled Plasmid DNA, and to significantly reduce the level of contaminating RNA, endotoxins and chromosomal DNA. In particular, a 100-fold reduction of chromosomal DNA contamination over that obtained with conventional techniques can be achieved through a single additional THAC step. Further improvements of THAC technology are possible, and we anticipate that this technique can be scaled up for integration into a full commercial-scale DNA production process.
Aurélie San Juan - One of the best experts on this subject based on the ideXlab platform.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:1 - ArticleIn the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA.
Journal of biomedical materials research. Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethylamine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc.
Grégory Ducrocq - One of the best experts on this subject based on the ideXlab platform.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:1 - ArticleIn the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA.
Journal of biomedical materials research. Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethylamine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc.
Didier Letourneur - One of the best experts on this subject based on the ideXlab platform.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:1 - ArticleIn the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA.
Journal of biomedical materials research. Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethylamine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies.
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Tubular cationized pullulan hydrogels as local reservoirs for Plasmid DNA
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Aurélie San Juan, Grégory Ducrocq, Hanna Hlawaty, Isabelle Bataille, Erwann Guénin, Didier Letourneur, Laurent J. FeldmanAbstract:In the present study, we measured the ability of various cationized pullulan tubular hydrogels to retain Plasmid DNA, and tested the ability of retained Plasmid DNA to transfect vascular smooth muscle cells (VSMCs). Cationized pullulans were obtained by grafting at different charge densities ethylamine (EA) or diethylaminoethyl-amine (DEAE) on the pullulan backbone. Polymers were characterized by elemental analysis, acid-base titration, size exclusion chromatography, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance. The complexation of cationized pullulans in solution with Plasmid DNA was evidenced by fluorescence quenching with PicoGreen. Cationized pullulans were then chemically crosslinked with phosphorus oxychloride to obtain tubular cationized pullulan hydrogels. Native pullulan tubes did not retain loaded Plasmid DNA. In contrast, the ability of cationized pullulan tubes to retain Plasmid DNA was dependent on both the amine content and the type of amine. The functional integrity of Plasmid DNA in cationized pullulan tubes was demonstrated by in vitro transfection of VSMCs. Hence, cationized pullulan hydrogels can be designed as tubular structures with high affinity for Plasmid DNA, which may provide new biomaterials to enhance the efficiency of local arterial gene transfer strategies. (c) 2007 Wiley Periodicals, Inc.