Influence of glioma cells on a new co-culture in vitro blood-brain barrier model for characterization and validation of permeability

Int J Pharm. 2015 Jul 25;490(1-2):94-101. doi: 10.1016/j.ijpharm.2015.05.027. Epub 2015 May 14.

Abstract

The blood-brain barrier plays an important role in protecting the brain from injury and diseases, but also restrains the delivery of potential therapeutic drugs for the treatment of brain illnesses, such as tumors. Glioma is most common cancer type of central nervous system in adults and the most lethal in children. The treatment is normally poor and ineffective. To better understand the ability of drug delivery systems to permeate this barrier, a blood-brain barrier model using human brain endothelial cells and a glioma cell line is herein proposed. The consistent trans-endothelial electrical values, immunofluorescence and scanning electronic microscopy showed a confluent endothelial cell monolayer with high restrictiveness. Upon inclusion of glioma cell line, the trans-endothelial electrical resistance decreased, with consequent increase of apparent permeability of fluorescein isothiocyanate dextran used as model drug, revealing a reduction of the barrier robustness. In addition, it was demonstrated a cell shape modification in the co-culture, with loss of tight junctions. The microenvironment of co-cultured model presented significant increase of of CCL2/MCP-1 and IL-6 production, correlating with the modulation of permeation. The results encourage the use of the proposed in vitro model as a screening tool when performing drugs permeability for the treatment of disorders among the central nervous system.

Keywords: Blood–brain barrier; Co-culture; In vitro model; Permeability studies; Trans-endothelial electrical resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood-Brain Barrier / metabolism
  • Blood-Brain Barrier / pathology*
  • Blood-Brain Barrier / physiology*
  • Brain / metabolism
  • Brain / pathology
  • Brain / physiology
  • Cell Line
  • Cell Line, Tumor
  • Cell Membrane Permeability / physiology*
  • Chemokine CCL2 / metabolism
  • Coculture Techniques / methods
  • Dextrans / metabolism
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology
  • Endothelial Cells / physiology
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / metabolism
  • Glioma / metabolism
  • Glioma / pathology*
  • Humans
  • Interleukin-6 / metabolism
  • Permeability
  • Tight Junctions / metabolism

Substances

  • CCL2 protein, human
  • Chemokine CCL2
  • Dextrans
  • Interleukin-6
  • fluorescein isothiocyanate dextran
  • Fluorescein-5-isothiocyanate