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Cellular vesicles (CVs) show promise for drug delivery, with hCMEC/D3-derived CVs demonstrating superior blood-brain barrier penetration and brain localization in vivo. Proteomics identified components influencing organotropism.

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Drug Delivery Systems

Background:

  • Cellular vesicles (CVs) are emerging as potential alternatives to exosomes for targeted drug delivery.
  • CVs can be engineered from various cell sources, including HEK-293, B16F10 melanoma, and hCMEC/D3 cells.

Purpose of the Study:

  • To characterize CVs for morphology, cytotoxicity, and cell uptake.
  • To evaluate the brain-targeting potential of CVs in vitro and in vivo.
  • To investigate the impact of cell origin and culture conditions on CV properties and brain localization.

Main Methods:

  • CVs were prepared using liposome technology methods and characterized by size, ζ-potential, and calcein loading/latency.
  • Dehydration-rehydration method was optimized for calcein loading.
  • In vitro blood-brain barrier (BBB) model (hCMEC/D3) and in vivo/ex vivo studies were used to assess brain targeting.
  • Proteomics was employed to analyze CV composition.

Main Results:

  • CVs ranged from 135-285 nm with negative ζ-potential.
  • The dehydration-rehydration method yielded optimal calcein loading and latency.
  • Pegylation improved CV integrity and reduced liver uptake, correlating with biodistribution predictions.
  • hCMEC/D3-derived CVs exhibited the highest in vitro BBB permeability and in vivo brain localization.
  • Different culture media for hCMEC/D3 cells affected CV interaction with brain cells and localization, linked to proteomic differences.

Conclusions:

  • CVs derived from hCMEC/D3 cells show significant potential for brain-targeted drug delivery.
  • CV integrity and biodistribution can be predicted using calcein leakage assays.
  • Proteomic analysis offers a method to identify CV components responsible for organotropism, enabling targeted delivery optimization.