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Article Dans Une Revue Bioelectrochemistry Année : 2024

Fibroblasts transfection by electroporation in 3D reconstructed human dermal tissue

Résumé

The understanding of the mechanisms involved in DNA electrotransfer in human skin remains modest and limits the clinical development of various biomedical applications, such as DNA vaccination. To elucidate some mechanisms of DNA transfer in the skin following electroporation, we created a model of the dermis using a tissue engineering approach. This model allowed us to study the electrotransfection of fibroblasts in a threedimensional environment that included multiple layers of fibroblasts as well as the self-secreted collagen matrix. With the aim of improving transfection yield, we applied electrical pulses with electric field lines perpendicular to the reconstructed model tissue. Our results indicate that the fibroblasts of the reconstructed skin tissue can be efficiently permeabilized by applied millisecond electrical pulses. However, despite efficient permeabilization, the transfected cells remain localized only on the surface of the microtissue, to which the plasmid was deposited. Second harmonic generation microscopy revealed the extensive extracellular collagen matrix around the fibroblasts, which might have affected the mobility of the plasmid into deeper layers of the skin tissue model. Our results show that the used skin tissue model reproduces the structural barriers that might be responsible for the limited gene electrotransfer in the skin.
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hal-04459793 , version 1 (15-02-2024)

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Géraldine Albérola, Elisabeth Bellard, Jelena Kolosnjaj-Tabi, Jorgan Guard, Muriel Golzio, et al.. Fibroblasts transfection by electroporation in 3D reconstructed human dermal tissue. Bioelectrochemistry, 2024, pp.108670. ⟨10.1016/j.bioelechem.2024.108670⟩. ⟨hal-04459793⟩
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