ENDOXY IN FLAME: ENDOTHELIAL AND IMMUNE CELL INTERACTIONS DURING BIOHYBRID LUNG APPLICATION

• Published in: International journal of artificial organs 2023-07, Vol.46 (7), p.434
• Main Authors: Cheremkhina, M, Krapp, S, Neullens, C, Ohl, K, Tenbrock, K, Jockenhoevel, S, Cornelissen, C G, Thiebes, A L
• Format: Article
• Language: English
• Subjects: Adhesion; Blood; CD31 antigen; Cell adhesion; Cell adhesion molecules; Cell interactions; Cell size; Cytology; Dynamical systems; E-selectin; Endothelial cells; Extracorporeal membrane oxygenation; Flow cytometry; Gas exchange; Immune system; Inflammation; Intercellular adhesion molecule 1; Leukocytes (mononuclear); Lipopolysaccharides; Lungs; Membranes; Microfluidics; Morphology; Oxygenation; Peripheral blood mononuclear cells; Polydimethylsiloxane; Shear stress; Umbilical vein; Vascular cell adhesion molecule 1; Von Willebrand factor; Wall shear stresses
• ISSN: 0391-3988; 1724-6040

Objectives: Extracorporeal membrane oxygenation (ECMO) therapy faces challenges such as limited hemocompatibility and an increased inflammatory reaction. One promising solution to these challenges is endothelialization of the gas exchange membrane. This study aims to investigate interaction between endothelial cells (ECs) and immune cells for biohybrid lung application, which has been poorly investigated so far. Methods: Human umbilical vein endothelial cells (HUVECs) were seeded on polydimethylsiloxane gas exchange membranes and cultured in a microfluidic system with wall shear stress of 20 dyn/cm2. After 24-hour pre-cultivation, peripheral blood mononuclear cells (PBMCs, 1.5 x 106 cells/mL) activated with lipopolysaccharides (LPS, 100 ng/mL) were added to the dynamic system for 24 hours. Cultures without LPS and/or PBMCs served as controls. Cell layer confluency was analyzed by immunohistochemical staining with CD31/PECAM-1 and von Willebrand factor. Flow cytometry analysis was performed on HUVECs to determine expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and E-selectin. Leukocyte adhesion assay was conducted to determine the number of adhered PBMCs. Results: Treatment with LPS-activated PBMCs caused a reduction of cell layer confluency and a change in HUVEC morphology towards a prolonged, aligned cell shape, as opposed to controls with non-activated PBMCs without LPS. Here, HUVECs showed typical cobblestone morphology. Flow cytometry analysis revealed significantly increased expression of VCAM-1, ICAM-1 and E-Selectin for HUVECs treated with LPS-activated PBMCs in contrast to controls. Increased adherence of PBMCs on HUVECs layer was observed after LPS-treatment. Conclusions: In conclusion, the study provides valuable insights into the interaction between ECs and blood immune cells during inflammation, which is relevant for biohybrid lung application. The results raise the question whether gas exchange membrane endothelialization is a feasible way to overcome current challenges of ECMO systems. Further studies are necessary to investigate this interaction and develop strategies to optimize the effectiveness of endothelialization for biohybrid lung application.