Abstract
To provide an alternative treatment option for patients with end-stage lung disease, we aim for biohybrid lung development (BHL) based on hollow fiber membrane (HFM) technology used in extracorporeal membrane oxygenators. For long-term BHL application, complete hemocompatibility of all blood-contacting surfaces is indispensable and can be achieved by their endothelialization. Indeed, albumin/heparin (AH) coated HFM enables initial endothelialization, but as inexplicable cell loss under flow conditions was seen, we assessed an alternative HFM coating using fibronectin (FN). Therefore, endothelial cell (EC) adherence and viability on both coated HFM were analyzed by fluorescence-based staining. Functional leukocyte and thrombocyte adhesion assays were performed to evaluate hemocompatibility, also in comparison to blood plasma coated HFM as a clinically relevant control. To assess monolayer resistance and EC behavior under clinically relevant flow conditions, a mock circulation setup was established, which also facilitates imitation of lung-disease specific blood gas settings. Besides quantification of flow-associated cell loss, endothelial responses towards external stimuli, like flow exposure or TNFα stimulation, were analyzed by qRT-PCR, focusing on inflammation, thrombus formation and extracellular matrix production. Under static conditions, both coated HFM enabled the generation of a viable, confluent, non-inflammatory and anti-thrombogenic monolayer. However, by means of homogenous FN coating, cell retention and physiologic gene regulation towards an improved hemocompatible-and extracellular matrix producing phenotype, was significantly superior compared to the inhomogeneous AH coating. In summary, our adaptable in-house FN coating secures the endothelial requirements for long-term BHL application and may promote monolayer establishment on all other blood contacting surfaces of the BHL (e.g., cannulae).
Highlights
Suffering from end-stage lung diseases (ELD), lung transplantation (LTx) is still the only curative therapy option
To provide an alternative treatment option for patients with end-stage lung disease, we aim for biohybrid lung development (BHL) based on hollow fiber membrane (HFM) technology used in extracorporeal membrane oxygenators
Fluorescence microscopy images revealed that both, AH (Figure 2A) and FN coated HFM (Figure 2B), enabled the generation of a viable and confluent endothelial monolayer, comprising endothelial cell (EC), which were interconnected via the cell junctional protein VE-cadherin (Figure 2C,D)
Summary
Suffering from end-stage lung diseases (ELD), lung transplantation (LTx) is still the only curative therapy option. ECMO application is limited to a few weeks only, as insufficient hemocompatibility of all artificial blood-contacting surfaces results in the adhesion of proteins and subsequent thrombocytes, initiating the formation of blood clots, which significantly impede the blood flow through the device [3], thereby decreasing its gas exchange efficiency [4,5]. Patients have to receive a meticulously controlled anti-coagulation therapy, which bears a high risk for severe and lethal hemorrhage [4]. These measures can only delay but not prevent the above-mentioned adverse blood-material interactions until device failure becomes inevitable. It is clear that there is an urgent need to develop a long-lasting alternative to ECMO, which can be used as bridge-to-lung transplantation (LTx) and as a final destination therapy, comparable to left ventricular assist devices for terminal heart failure [8]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
