Figure 1.

Model optimization. (a) Optimization of basement membrane substitute. LY permeation across bEnd3 monolayer grown on the top of polycarbonate transwell coated with 150 or 300 μg/ml rat-tail collagen type I, 30 or 60 μg/ml fibronectin, and a mixture of 150 μg/ml rat-tail collagen and 30 μg/ml fibronectin. LY permeation was represented as permeation coefficient (Pc). Compared to other tested coatings, in the presence of cells, fibronectin at 30 μg/ml offered the lowest Pc value indicative of restricted transport due to formation of a tight monolayer, and in the absence of cells fibronectin did not act as a barrier to LY transport. Data represented as mean±SEM for 9 replicates from 3 independent experiments. * P<0.05, ** P<0.01 values compared to control, # P<0.05 compared to collagen at 150 μg/ml.(b) Time-dependent changes in bEnd3 cells growth (open circles) and LY permeation across the bEnd3 cells-based BBB model (open triangles). Growth pattern of bEnd3 cells after seeding indicates formation of confluent monolayer on day 3 post-seeding. On the other hand, LY permeation started to decrease from day 1 and continue to decrease until day 4 where it remains contestant until day 6. LY permeation change indicates formation of tight junction between cells in confluent bEnd3 monolayer. Data represented as mean±SEM for 6 replicates from 2 independent experiments (* p < 0.05).