Abstract
The blood brain barrier (BBB) is an anatomical structure composed of endothelial cells, basement membrane and glia; preventing drugs and chemicals from entering the brain. Our aim is to engineer an in vitro BBB model in order to facilitate neurological drug development that will ultimately benefit patients. Tissue engineering approaches are useful for the generation of an in vitro BBB model. Our experimental approach is to mimic the anatomical structure of the BBB on polyester terphthalate (PET) cell culture inserts. Endothelial cells derived from brain capillaries, different peripheral blood vessels and epithelial cells (MDCK) were cultured on the apical side of the filter. Different concentrations of thrombin were applied on compact monolayer of MDCK. Physiological functions of this BBB model was evaluated by measuring the transendothelial resistance (TEER) using an EndOhM (TM) electrode. The epithelial cytoskeletal organization was observed by staining with BBZ-phalloidin. Epithelial monolayer formation and later distortion by thrombin was confirmed by fluorescence microscopy. Measurements of the TEER generated values up to 2020 ohm/cm(2). A dose response of thrombin was observed, showing the permeability changes in the epithelial cells (MDCK). A relationship between permeability values TEER and cytoskeletal organization was observed.