Towards Biohybrid Lung Development? Inflammatory Conditions Disrupt Endothelial Layer Integrity on Gas Exchange Membranes

Abstract

AbstractNext to limited hemocompatibility, inflammation and sepsis are frequent complications during extracorporeal membrane oxygenation. Endothelialization of gas exchange membranes has been proposed to overcome these limitations and its general feasibility was demonstrated. However, these investigations focused on application of biohybrid devices under standard culture conditions neglecting patients’ inflammatory status in future application. In this study, we investigate endothelial layer behavior on gas exchange membranes under inflammatory conditions in a microfluidic model system using immunocytochemistry, scanning electron microscopy, flow cytometry and qPCR. While co-culture with peripheral blood mononuclear cells (PBMCs) does not change endothelial layer integrity, confluence of endothelial cells is substantially reduced during inflammation via LPS-activated PBMCs. Cell adhesion molecules were increasingly expressed under inflammatory conditions, consistent with an increased leukocyte adhesion. An upregulation of several genes linked to inflammation is observed: ICAM-1, VCAM-1, E-Selectin, IL6, IL8, IL10, and MCP-1. Our findings suggest that endothelial cells may struggle to maintain layer integrity within a biohybrid device when exposed to inflammatory conditions. This raises the question of whether endothelialization is an effective advancement of current technologies considering inflammation in patients. Yet the presented setup qualifies as sepsis in vitro model replicating the physiological vascular leak phenomenon to aid future investigations in biohybrid lung research.