Biomedical subjects

Justin Teissie

Publications and source records attributed to Justin Teissie.

4 recordsLinked to original sources

PubMed2006

New anti angiogenesis developments through electro-immunization: optimization by in vivo optical imaging of intradermal electro gene transfer.

Direct application of high voltage electric pulses of milliseconds duration to the skin of a mouse enhances in vivo intradermal delivery of injected therapeutic molecules such as DNA. The efficacy of gene transfer and expression is dependent on electrical parameters. DNA electrotransfer in tissues increases the associated DNA expression vaccine potency. This protocol is called "electro-immunization". In the present study, we report a new strategy for optimizing electro-immunization. In vivo fluorescence imaging was used to detect the expression of a fluorescent protein (DsRed) and therefore allowed rapid optimization of the protocol. In vivo electrogenetransfer in the skin was well tolerated and DsRed expression was followed for over 2 weeks. Expression was voltage dependent under our conditions. Parameters were selected giving the highest level of expression. Under these optimized conditions, electrotransfer of a plasmid encoding VEGF was evaluated for its immune response as a gene therapy of interest involved in anti-angiogenic strategies. Anti VEGF 165 antibodies in sera of mice were evaluated by ELISA and compared to those obtained after conventional immunization. Comparable titres of antibodies were obtained in both groups. An IgG2a predominance was found in mice immunized with the plasmid whereas a IgG1 predominance was observed in mice immunized classically. Skin electro-immunization is therefore shown as a good route for DNA immunization for anti-angiogenesis concern.

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PubMed2003

Finite-element modeling of needle electrodes in tissue from the perspective of frequent model computation.

Information about electric field distribution in tissue is very important for effective electropermeabilization. In heterogeneous tissues with complex geometry, finite-element (FE) models provide one of alternative sources of such information. In the present study, modeling of needle electrode geometry in the FE model was investigated in order to determine the most appropriate geometry by considering the need for frequent FE model computation present in electroporation models. The 8-faceted needle electrode geometry proposed--determined on a model with a single needle electrode pair by means of criteria function--consisted of the weighted sum of relative difference between measured and computed total current, the relative difference in CPU time spent on solving model, and the relative difference in cross section surface of electrodes. Such electrode geometry was further evaluated on physical models with needle arrays by comparison of computed total current and measured current. The agreement between modeled and measured current was good (within 9% of measurement), except in cases with very thin gel. For voltage above 50 V, a linear relationship between current and voltage was observed in measurements. But at lower voltages, a nonlinear behavior was detected resulting from side (electrochemical) effects at electrode-gel interface. This effect was incorporated in the model by introducing a 50-V shift which reduced the difference between the model and the measurement to less than 3%. As long as material properties and geometry are well described by FE model, current-based validation can be used for a rough model validation. That is a routine assay compared with imaging of electric field, which is otherwise employed for model validation. Additionally, current estimated by model, can be preset as maximum in electroporator in order to protect tissue against damage.

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PubMed2002

Cell hybridization by electrofusion on filters.

Electric field pulses induce permeabilization and associated fusogenicity in cell membranes. Electrofusion of cells is usually performed in two steps: the first is the creation of close intercellular contacts; the second is an application of electric pulses that induces membrane fusion. Very large cell contacts can be obtained by a filter aspiration method. A cell monolayer is created by controlled suction on biocompatible filter. No spontaneous fusion results. Just after filtration, electrofusion is obtained by field pulses applied parallel to the filter. Cell viability is not strongly affected and cells recover their spherical shape in the minute time range after filtration. The electrical parameters, the cell density, and the flow rate control fusion. Fusion is obtained with cells of different origins with very different adhesion properties. Hybrid cells are easily formed. This approach appears to be a very efficient method for cell hybridization with an easy-to-use protocol.

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PubMed2002

Direct visualization at the single-cell level of electrically mediated gene delivery.

Electropermeabilization is one of the nonviral methods successfully used to transfer genes into living cells in vitro and in vivo. Although this approach shows promise in the field of gene therapy, very little is known about the basic processes supporting DNA transfer. The present investigation studies this process at the single-cell level by using digitized fluorescence microscopy. Permeabilization is a prerequisite for gene transfer. Its assay by propidium-iodide (PI) penetration shows that it occurs at the sides of the cell membrane facing the two electrodes, whereas fluorescently labeled plasmids only interact with the electropermeabilized side of the cell facing the cathode. The plasmid interaction with the electropermeabilized part of the cell surface results in the formation of localized aggregates. These membrane-associated spots are formed only when pulses with a longer duration than a critical value are applied. These complexes are formed within 1 s after the pulses and cannot be destroyed by pulses of reversed polarities. They remain at the membrane level up to 10 min after pulsing. Although freely accessible to DNA dye (TOTO-1) 1 min after the pulses, they are fully protected when the addition takes place 10 min after. They diffuse in the cytoplasm 30 min after pulses and are present around the nucleus 24 h later.

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