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<p>Nanoparticles of various sizes and materials are widely used for targeted intracellular delivery in mammalian cell models. However, endocytic uptake often leads to endosomes embedding, reducing delivery efficiency, especially for larger particles. Electroporation offers a direct means of accessing the cytosol, bypassing this limitation. Here, we employ a microchip featuring integrated capacitive microelectrodes to perform spatially selective, rapid and efficient electroporation of 200&#xa0;nm fluorescent nanoparticles into adherent CHO-K1 epithelial cells. Operating at low voltage amplitudes, the system minimizes cellular damage while maintaining high delivery efficiency. We confirm effective cytoplasmic delivery and preferential perinuclear localization of internalized nanoparticles, with minimal impact on cell viability. Through a biophysical two-compartment model of the cell-microelectrode system, we simulate and explain the dependency of delivery efficiency of nanoparticles from the characteristics of electroporation stimuli and from cell geometry. The approach represents a promising platform for applications such as nanoparticle-mediated drug delivery and high-throughput screening even with particles approaching or exceeding 100&#xa0;nm in diameter.</p>

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Fast and efficient intracellular delivery of large size nanoparticles into mammalian cells by in situ electroporation

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月16日

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  • Marta Maschietto,
  • Enzo Cancedda,
  • Daniele Andrean,
  • Stefano Vassanelli

摘要

Nanoparticles of various sizes and materials are widely used for targeted intracellular delivery in mammalian cell models. However, endocytic uptake often leads to endosomes embedding, reducing delivery efficiency, especially for larger particles. Electroporation offers a direct means of accessing the cytosol, bypassing this limitation. Here, we employ a microchip featuring integrated capacitive microelectrodes to perform spatially selective, rapid and efficient electroporation of 200 nm fluorescent nanoparticles into adherent CHO-K1 epithelial cells. Operating at low voltage amplitudes, the system minimizes cellular damage while maintaining high delivery efficiency. We confirm effective cytoplasmic delivery and preferential perinuclear localization of internalized nanoparticles, with minimal impact on cell viability. Through a biophysical two-compartment model of the cell-microelectrode system, we simulate and explain the dependency of delivery efficiency of nanoparticles from the characteristics of electroporation stimuli and from cell geometry. The approach represents a promising platform for applications such as nanoparticle-mediated drug delivery and high-throughput screening even with particles approaching or exceeding 100 nm in diameter.