Recapitulation of 3D Cylindrical Human Blood-Brain Barrier in vitro by Brain and Blood Vessel Tissue Specific Bioinks and 3D Bioprinting Technology

Abstract

Neuroinflammation-associated diseases have affected the health and quality of life of people globally. The major regulator of neuroinflammation is the blood-brain barrier (BBB), which controls the molecular transport from brain vasculature to the central nervous system. Since the BBB is a 3D complex structure consisting of various cell types, 2D cell culture models or animal models are difficult to use for in-depth mechanistic studies of neuroinflammation. Therefore, it is necessary to recapitulate tissue-specific intrinsic function and 3D cylindrical structure of the human BBB in vitro. Previous researches demonstrated that decellularized extracellular matrix (dECM) has advantage in providing tissue specific micro-environmental cues[1,2]. In addition, previous study developed cylindrical Bio-Blood-Vessels using coaxial-based 3D cell printing strategy[3]. Here, we conducted characterization and validation of brain-derived dECM (BdECM) and blood vessel-derived dECM (VdECM), and coaxial-based cylindrical fiber printing process optimization. Proteomic analysis revealed that BdECM and VdECM have target tissue specific ECM proteins. Furthermore, it is shown that the mixture of BdECM and VdECM can provide more stable environment for BBB-constructing cells. Finally, we fabricated cylindrical BBB tissue using coaxial-based printing process and validated its potential barrier function. Later, this study will be the basis for developing BBB-on-a-Chip investigating the mechanism of neuroinflammation.