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<p>Cell permeabilization techniques are widely used to enable intracellular delivery of therapeutic molecules. Although these methods show promise for drug delivery and gene therapy, controlling and quantifying molecular transport at the nanoscale remains a significant challenge due to the structural complexity of mammalian cells. Multivesicular vesicles (MVVs), consisting of a large outer vesicle encapsulating multiple smaller vesicles, offer a simplified biomimetic platform that captures key aspects of cellular compartmentalization of mammalian cells. In this study, we employed finite element method (FEM)-based simulations using COMSOL Multiphysics to investigate passive molecular transport through permeabilized MVVs. The model incorporated biologically relevant pore sizes and three commonly used fluorescent probes of increasing molecular size: Calcein, Texas-red dextran 3000 (TRD-3k), and TRD-10k. The results reveal size-dependent accumulation kinetics. Additionally, transport efficiency was enhanced with increased pore diameter, indicating that both molecular and structural parameters influence delivery outcomes. Importantly, the simulations demonstrate that compartmentalized vesicle structures cause transport to vary across space and time. The model successfully matches experimental results seen in permeabilized vesicles and expands them to more complex nested vesicle systems. These findings highlight the role of vesicle architecture in shaping transport outcomes and provide a computational framework for exploring compartmentalization effects relevant to intracellular delivery strategies.</p>

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Simulation of intracellular delivery through permeabilized multivesicular vesicles

  • 期刊论文

  • 发表日期：

    2026年1月21日

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  • Shah Sajnin Anna,
  • Shahariar Emon,
  • Md. Asaduzzaman,
  • Shovon Saha,
  • Md. Atikur Rahman,
  • Mohammad Abu Sayem Karal,
  • Md Lokman Hossen,
  • Samiron Kumar Saha,
  • Hiromitsu Takaba,
  • Md. Akhtaruzzaman,
  • Md. Khorshed Alam

摘要

Cell permeabilization techniques are widely used to enable intracellular delivery of therapeutic molecules. Although these methods show promise for drug delivery and gene therapy, controlling and quantifying molecular transport at the nanoscale remains a significant challenge due to the structural complexity of mammalian cells. Multivesicular vesicles (MVVs), consisting of a large outer vesicle encapsulating multiple smaller vesicles, offer a simplified biomimetic platform that captures key aspects of cellular compartmentalization of mammalian cells. In this study, we employed finite element method (FEM)-based simulations using COMSOL Multiphysics to investigate passive molecular transport through permeabilized MVVs. The model incorporated biologically relevant pore sizes and three commonly used fluorescent probes of increasing molecular size: Calcein, Texas-red dextran 3000 (TRD-3k), and TRD-10k. The results reveal size-dependent accumulation kinetics. Additionally, transport efficiency was enhanced with increased pore diameter, indicating that both molecular and structural parameters influence delivery outcomes. Importantly, the simulations demonstrate that compartmentalized vesicle structures cause transport to vary across space and time. The model successfully matches experimental results seen in permeabilized vesicles and expands them to more complex nested vesicle systems. These findings highlight the role of vesicle architecture in shaping transport outcomes and provide a computational framework for exploring compartmentalization effects relevant to intracellular delivery strategies.