Influence of Aerosolization on Endothelial Cells for Efficient Cell Deposition in Biohybrid and Regenerative Applications

• Published in: Micromachines (Basel) Vol. 14; no. 3; p. 575
• Main Authors: Cheremkhina, Maria, Klein, Sarah, Babendreyer, Aaron, Ludwig, Andreas, Schmitz-Rode, Thomas, Jockenhoevel, Stefan, Cornelissen, Christian G, Thiebes, Anja Lena
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 28-02-2023; MDPI
• Subjects: Aerosols; Apoptosis; biohybrid lung; cell aerosolization; cell atomization; cell seeding; Endothelial cells; Endothelium; EndOxy; Extracorporeal membrane oxygenation; Flow velocity; Gas exchange; Gene expression; human umbilical vein endothelial cells (HUVECs); Kinases; Lungs; Membranes; Microfluidics; Necrosis; Shear stress; Software; Spraying; Substrates; Surgical apparatus & instruments; Survival; Umbilical cord; Veins & arteries; Von Willebrand factor; Germany; Austria; cell seeding; cell atomization; biohybrid lung; cell aerosolization; human umbilical vein endothelial cells (HUVECs); EndOxy
• ISSN: 2072-666X; 2072-666X
• DOI: 10.3390/mi14030575

The endothelialization of gas exchange membranes can increase the hemocompatibility of extracorporeal membrane oxygenators and thus become a long-term lung replacement option. Cell seeding on large or uneven surfaces of oxygenator membranes is challenging, with cell aerosolization being a possible solution. In this study, we evaluated the endothelial cell aerosolization for biohybrid lung application. A Vivostat
system was used for the aerosolization of human umbilical vein endothelial cells with non-sprayed cells serving as a control. The general suitability was evaluated using various flow velocities, substrate distances and cell concentrations. Cells were analyzed for survival, apoptosis and necrosis levels. In addition, aerosolized and non-sprayed cells were cultured either static or under flow conditions in a dynamic microfluidic model. Evaluation included immunocytochemistry and gene expression via quantitative PCR. Cell survival for all tested parameters was higher than 90%. No increase in apoptosis and necrosis levels was seen 24 h after aerosolization. Spraying did not influence the ability of the endothelial cells to form a confluent cell layer and withstand shear stresses in a dynamic microfluidic model. Immunocytochemistry revealed typical expression of CD31 and von Willebrand factor with cobble-stone cell morphology. No change in shear stress-induced factors after aerosolization was reported by quantitative PCR analysis. With this study, we have shown the feasibility of endothelial cell aerosolization with no significant changes in cell behavior. Thus, this technique could be used for efficient the endothelialization of gas exchange membranes in biohybrid lung applications.