The effect of non-invasive dermal electroporation on skin barrier function and skin permeation in combination with different dermal formulations
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The aim of this research was to investigate the effect of non-invasive dermal electroporation (EP) on the barrier function of the skin and on the permeation of a model macromolecule in combination with different dermal formulations. Skin samples were treated with non-invasive dermal EP treatment for 2 min. Firstly, the effect of EP on the barrier function of the skin was examined on mouse skin in vivo by measuring transepidermal water loss (TEWL). Then, the effect of EP on human skin permeation was investigated ex vivo under a fluorescence microscope in combination with different dermal formulations. The human skin was treated with a solution, a hydrogel, and a film-forming system (FFS) containing 4 kDa fluorescein isothiocyanate-dextran (FITC-dextran) with or without EP. The increased fluorescence intensity shows the presence of FITC-dextran in the skin layers. The results showed, that the TEWL values increased rapidly after the treatment, and it took approximately 5 min to be restored. The results of permeation experiments showed that just slight permeation of FITC-dextran could be noticed from any formulation without EP; however, the permeation from the solution and the FFS increased highly in combination with EP. The EP decreased the barrier function of the skin reversibly and the structure of SC was restored in a short time after the treatment. FITC-dextran, as a macromolecule, can just slightly permeate into the skin with passive diffusion. EP could increase the permeation rate of FITC-dextran remarkably compared to the control treatments; however, the composition of the formulations has a great influence on the permeation.Keywords:
Transepidermal water loss
Barrier function
Fluorescein isothiocyanate
Human skin
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1) The globulin fractions of antisera representing bacterial, viral, mycotic agents and antiglobulin fractions were labeled with 2 derivatives of fluorescein amine by 3 methods. 2) Fluorescein isothiocyanate was shown to be superior in stability, ease of conjugation, and degree of fluorescence. This direct method of adding the dye to a dilute buffered antiserum eliminates the need for organic reagents which may denature the protein.
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Artificial reduction of abnormal transepidermal water loss (TEWL) is considered to improve skin diseases associated with a defective barrier function. Treatment of the skin with moisturizers is also known to influence skin barrier function. Whether or not differences in occlusion between creams contribute to their effects on the skin barrier function is unknown.To investigate the long-term effects of a semipermeable membrane on the skin barrier function in normal skin. In addition, the occlusive properties of two creams were studied.The study was randomized, controlled and evaluator-blind using measurement of TEWL and skin susceptibility to sodium lauryl sulphate as indicators of skin barrier function.Coating of the skin with a silicone membrane for 23 h per day for 3 weeks improved skin barrier function, whereas no significant changes were found after using the membrane for 8 h per day.Differences between creams in terms of their effect on skin barrier function cannot be solely explained by their occlusive properties.
Transepidermal water loss
Skin Barrier
Barrier function
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Attempts to increase the sensitivity of fluorescein-based fluorescence immunoassays by using multiple labelling have generally been unsuccessful because of concentration quenching. We have labelled antibodies to human immunoglobulin G with multiple fluorescein fluorophores attached by means of a disulphide linkage: this linkage can be rapidly and easily broken by treatment with dithiothreitol, allowing fluorescein to be released from the antibody and measured in free solution. Application of this technique to a fluorescence labelled immunosorbent assay for antibodies to the human immunodeficiency virus gave an approximately 20-fold increase in signal compared with an equivalent assay using fluorescein isothiocyanate.
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Objective: To build a fluorescein labeling method to detect the bingding of aminopeptidase N inhibitors with tumor cells. Methods: Aminopeptidase N inhibitors LYRM03 and Bestatin were labeled with fluorescein isothiocyanate(FITC), and fluorescein-labeled compounds of FITC-LYRM03 and FITC-Bestatin were purified. The relationship between binding activities to tumor cells and inhibitory activities of aminopeptidase N inhibitors was detected by using fluorescence microscope and flow cytometer. Results: Compounds of LYRM03 and Bestatin had inhibitory activities to aminopeptidase N of tumor cells. Fluorescein-labeled FITC-LYRM03 and FITC-Bestatin had different binding activities to tumor cells. Conclusion: There was consistency between binding activities of tumor cells and inhibitory activities of aminopeptidase N inhibitors.
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Nanoparticles with a diameter of between 100 and 800 nm can be made from gelatin or albumin using a desolvation and hardening technique. Fluorescein isothiocyanate (FITC) can be bound to the surface of such nanoparticles. The conjugated gelatin nanoparticles were phagocytozed by some experimental tumour lines.
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Abstract Fluorescein isothiocyanate has been used to label normal or tumor cells in order to study their migration in vivo. This requires that any transfer of fluorescence to neighbouring cells be carefully ruled out. The aim of the present report is to demonstrate the possibility of a transfer of fluorescein or fluorescein‐bound molecules between untreated and labeled cells. When normal rat thymocytes were co‐incubated with labeled cells (about 5 × 10 6 fluorescein molecules/cell) under continuous agitation or exposed to supernatants of these labeled cells, they bound an average of 10 4 fluorescein molecules. When the incubation was done on cell pellets after centrifugation, this transfer was increased tenfold. Hence, intercellular molecule exchange may occur in the absence of any specific interaction.
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