Modelling Inflammatory Endothelial Dysfunction: A Human In Vitro Platform for Translational Research

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Systemic inflammation presents a significant challenge to the long-term function of biohybrid implants. While endothelialisation of biohybrid implants has been shown to improve device hemocompatibility, its feasibility under the influence of patients’ inflammatory status remains largely unexplored. To investigate this, we developed a controlled in vitro model which allows to study endothelial dysfunction under inflammatory stress. Endothelial cells were cultured on polydimethylsiloxane under physiological shear stress and exposed to lipopolysaccharide (LPS)-activated peripheral blood mononuclear cells (PBMCs), simulating inflammatory conditions. Endothelial morphology and confluence was assessed using immunohistochemistry and scanning electron microscopy. Leukocyte adhesion was evaluated directly as well as indirectly, using flow cytometry to analyse cell adhesion molecules. Quantitative PCR was used for gene expression analysis of inflammatory mediators. Notably, neither LPS nor PBMCs alone induced endothelial disruption, whereas their combination significantly impaired endothelial confluence: Inflammatory activation led to substantial loss of endothelial confluence, increased leukocyte adhesion, and elevated expression of adhesion molecules ICAM-1, VCAM-1, and E-selectin. Gene expression analysis highlights the upregulation of inflammatory mediators, such as IL-6, IL-8, IL-10, and MCP-1. This study underscores the challenges of implementing endothelialisation in biohybrid devices, particularly in patients with systemic inflammation. By considering translational hurdles, this work contributes to the development of clinically viable biohybrid constructs and highlights the importance of considering inflammatory dynamics when designing next-generation implants.

Authors 11

1. Maria Cheremkhina Aachen Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering

   RWTH Aachen University

   Affiliation as printed

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University, Forckenbeckstraße 55, 52074 Aachen, Germany

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University

2. Aaron Babendreyer Aachen Institute of Molecular Pharmacology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Molecular Pharmacology, University Hospital RWTH Aachen, Wendlingweg 2, 52074 Aachen, Germany

   Institute of Molecular Pharmacology, University Hospital RWTH Aachen

3. Christopher T. Neullens Aachen Division of Pediatric Immunology Department of Pediatrics

   RWTH Aachen University

   Affiliation as printed

   Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany

   Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University

4. Susanne Krapp Aachen Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering

   RWTH Aachen University

   Affiliation as printed

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University, Forckenbeckstraße 55, 52074 Aachen, Germany

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University

5. Kim Ohl Aachen Division of Pediatric Immunology Department of Pediatrics

   RWTH Aachen University

   Affiliation as printed

   Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany

   Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University

6. Stephan Ruetten Aachen Institute of Pathology, Electron Microscopy Facility Division of Pediatric Immunology Department of Pediatrics

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Pathology, Electron Microscopy Facility, RWTH Aachen University Hospital, Aachen, Germany

   Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University

7. Andreas Ludwig Aachen Institute of Molecular Pharmacology Institute of Pathology, Electron Microscopy Facility

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Molecular Pharmacology, University Hospital RWTH Aachen, Wendlingweg 2, 52074 Aachen, Germany

   Institute of Pathology, Electron Microscopy Facility, RWTH Aachen University Hospital

8. Christian Gabriel Cornelissen Aachen Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering Department of Pneumology and Internal Intensive Care Medicine Medical Clinic V Institute of Molecular Pharmacology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University, Forckenbeckstraße 55, 52074 Aachen, Germany

   Department of Pneumology and Internal Intensive Care Medicine, Medical Clinic V, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany

   Institute of Molecular Pharmacology, University Hospital RWTH Aachen

9. Stefan Jockenhoevel Aachen Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering Department of Pneumology and Internal Intensive Care Medicine Medical Clinic V

   Maastricht University · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University, Forckenbeckstraße 55, 52074 Aachen, Germany

   Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University

   Department of Pneumology and Internal Intensive Care Medicine, Medical Clinic V, University Hospital RWTH Aachen

10. Klaus Tenbrock Aachen Division of Pediatric Immunology Department of Pediatrics Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering

    RWTH Aachen University

    Affiliation as printed

    Division of Pediatric Immunology, Department of Pediatrics, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany

    Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University

11. Anja Lena Thiebes corresponding Aachen Department of Biohybrid and Medical Textiles (BioTex) Institute of Applied Medical Engineering

    Maastricht University · RWTH Aachen University

    Affiliation as printed

    Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University, Forckenbeckstraße 55, 52074 Aachen, Germany

    Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus

    Department of Biohybrid & Medical Textiles (BioTex), Institute of Applied Medical Engineering, Helmholtz Institute Aachen, RWTH Aachen University