A novel in vitro blood-brain barrier model using 3D bioprinter: A pilot study

Abstract

3D bioprinters allow the creation of functional tissues using various cell lines. The blood-brain barrier (BBB) is crucial for maintaining homeostasis in the brain, acting as a protective interface between the cerebral blood vessels and the brain parenchyma. Advancements in tissue engineering have led to the development of in vitro BBB models. However, model accuracy and reproducibility still require improvements. Thus, we hypothesized that producing BBB models using a 3D bioprinter is a novel method to overcome these limitations. We tested the maintainability of habitable conditions for cells and the durability of the 3D-bioprinted BBB model. We printed four different BBB models using a bio-ink composed of Matrigel base and CPA47 endothelial cells: single- and double-layered square and circular scaffolds. The double-layered square scaffold fostered optimal BBB functions as it most effectively maintained a fixed shape and initial cell positions. When A172 glioblastoma cells were placed in the empty square spaces of the square BBB scaffold, most cells were alive and healthy after 96 hours. Although A172 cells remained in their initial cell positions until the first 48 hours, we observed movement across the Matrigel barrier when checked at 96 hours. At 48 hours, A172 cells showed no movement from their initial positions; however, by 96 hours, we observed clear movement of A172 cells across the Matrigel barrier. Due to its relative cost-effectiveness and high reproducibility, our BBB model has several applications, especially for studying brain cancer metastasis past the BBB and developing potential corresponding clinical therapeutics.