PROJECT SUMMARY – Core C New pre-clinical models of both the airway and gastrointestinal epithelium, especially those that adequately reflect relevant human 3D physiology and disease pathophysiology, are desperately needed to elucidate disease mechanisms and identify avenues for treatment. The overall objective of the Engineering MicroEnvironment Core (EMEC) is to provide the group of Biomimetic Collaborative Research Center (BCRC) investigators with biomaterial and fluidic chamber platforms and additional enabling technologies to improve human gastrointestinal and lung systems for the studies proposed in Projects 1-3 and the Human Biomimetic Scientific Core (HBSC, Core B). These biomimetic systems are designed to replicate key aspects of the epithelial cells’ 3D physiological and physical environment. These platforms will utilize the biomaterial and tissue engineering technologies that we established during our original NAMSED funding, and will also build upon these technologies to expand our capabilities to answer questions about the role of the host mucus layer, cell physical microenvironment, and cell communities in intestinal and lung infections. The service component of the EMEC will be to provide engineering tools, including (1) preparing “TransWell Trough” systems to apply flow to co- cultures of anatomically-distinct epithelial cells, (2) fabricating tissue engineering/biomaterial platforms to support intestinal or lung epithelial cell cultures, (3) fabricating millifluidic perfusion chambers (mPC) for flow across intestinal epithelial cells ± pathogens, (4) fabricating and maintaining calibrated stocks of oxygen-sensing hydrogel-based microparticles, (5) 3D printing of molds and other components of the culture systems being fabricated, (6) quantifying tissue and biofluid mechanical behavior to prepare in vitro models with physiologically faithful material properties, (7) computational modeling of fluid dynamics and oxygen transport in culture systems, and (8) transferring technology through training group members and personnel at other funded U19s. The development component of EMEC will enhance the previously tested culture systems to mimic the complexity of the 3D host environment in the proposed studies, through (1) developing a modification of the TransWell Trough model with dual flow, (2) modifying the hydrogels to enable 3D encapsulation of immune and neural cells for co-culture studies, (3) developing a modified mPC system to grow the epithelial cells atop a biomimetic hydrogel surface, and (4) developing customized mucosal mimics to facilitate screening of host mucus-pathogen interactions. Providing these platforms, tools, and services through a central core will save time, effort, and costs, accelerate the rate of discovery, and enable comparison of results across Projects whenever possible. The EMEC will be consultative and responsive to needs of the individual Projects, which may change as the research proceeds and as the over...