Development of a bioreactor system for pre‐endothelialized cardiac patch generation with enhanced viscoelastic properties by combined collagen I compression and stromal cell culture

Journal of Tissue Engineering and Regenerative Medicine, vol. 14, pp. 1749–1762

Abstract

Treatment of terminal heart failure still poses a significant clinical problem. Cardiac tissue engineering could offer autologous solutions for the replacement of nonfunctional myocardial tissue. So far, soft matrix construction and missing large-scale prevascularization prevented the application of sizeable cardiac repair patches. We developed a novel bioreactor system for semi-automatic compression of a collagen I hydrogel applying 16 times higher pressure than in previous studies. Resistance towards compression stress was investigated for multiple cardiac-related cell types. For scaffold prevascuarization, a tubular cavity was imprinted during the compaction process. Primary cardiac-derived endothelial cells (ECs) were isolated from human left atrial appendages (HLAAs) and characterized by fluorescence-activated cell sorting (FACS) and immunocytology. EC were then seeded into the preformed channel with dermal fibroblasts as interstitial cell component of the fully cellularized patch. After 8 days of constant perfusion culture within the same bioreactor, scaffold dynamic modulus and cell viability were analyzed. Endothelial proliferation and vessel maturation were examined by immunohistochemistry and transmission electron microscopy. Our design allowed for scaffold production and dynamic culture in a one-stop-shop model. Enhanced compression and cell-mediated matrix remodeling induced a significant increase in scaffold stiffness while ensuring excellent cell survival. For the first time, we could isolate HLAA-derived EC with proliferative potential. ECs within the central channel proliferated during flow culture, continuously expressing endothelial markers (CD31) and displaying basal membrane synthesis (collagen IV, ultrastructural analysis). After 7 days of culture, a complete endothelial monolayer could be observed. Covering cells aligned themselves in flow direction and developed mature cell-cell contacts.

Authors 7

1. Carolin Krziminski

   Universitätsklinikum Würzburg

   Affiliation as printed

   Chair of Tissue Engineering and Regenerative Medicine Wuerzburg University Hospital Wuerzburg Germany

   Chair of Tissue Engineering and Regenerative Medicine, Wuerzburg University Hospital, Wuerzburg, Germany

2. Sebastian Kammann Aachen Fraunhofer Institute for Production Technology (IPT) Aachen Germany Fraunhofer Institute for Production Technology

   Fraunhofer Institute for Production Technology IPT · Universitätsklinikum Würzburg

   Affiliation as printed

   Chair of Tissue Engineering and Regenerative Medicine Wuerzburg University Hospital Wuerzburg Germany

   Fraunhofer Institute for Production Technology (IPT) Aachen Germany

   Chair of Tissue Engineering and Regenerative Medicine, Wuerzburg University Hospital, Wuerzburg, Germany

   Fraunhofer Institute for Production Technology (IPT), Aachen, Germany

3. Jan Hansmann

   Technical University of Applied Sciences Würzburg-Schweinfurt · Fraunhofer Institute for Silicate Research

   Affiliation as printed

   Faculty Electrical Engineering University for Applied Sciences Wuerzburg‐Schweinfurt Schweinfurt Germany

   Translational Center Regenerative Therapies Fraunhofer Institute for Silicate Research (ISC) Wuerzburg Germany

   Faculty Electrical Engineering, University for Applied Sciences Wuerzburg-Schweinfurt, Schweinfurt, Germany

   Translational Center Regenerative Therapies, Fraunhofer Institute for Silicate Research (ISC), Wuerzburg, Germany

4. Frank Edenhofer

   Universität Innsbruck · University of Würzburg · Otto-von-Guericke-Universität Magdeburg

   Affiliation as printed

   Institute of Anatomy and Cell Biology University of Wuerzburg Wuerzburg Germany

   Institute of Molecular Biology University of Innsbruck Innsbruck Austria

   Research Center Dynamic Systems: Systems Engineering Otto‐von‐Guericke‐University Magdeburg Germany

   Institute of Anatomy and Cell Biology, University of Wuerzburg, Wuerzburg, Germany

   Institute of Molecular Biology, University of Innsbruck, Innsbruck, Austria

   Research Center Dynamic Systems: Systems Engineering, Otto-von-Guericke-University, Magdeburg, Germany

5. Gudrun Dandekar

   Universitätsklinikum Würzburg · Fraunhofer Institute for Silicate Research

   Affiliation as printed

   Chair of Tissue Engineering and Regenerative Medicine Wuerzburg University Hospital Wuerzburg Germany

   Translational Center Regenerative Therapies Fraunhofer Institute for Silicate Research (ISC) Wuerzburg Germany

   Chair of Tissue Engineering and Regenerative Medicine, Wuerzburg University Hospital, Wuerzburg, Germany

   Translational Center Regenerative Therapies, Fraunhofer Institute for Silicate Research (ISC), Wuerzburg, Germany

6. Heike Walles

   Universität Innsbruck · Universitätsklinikum Würzburg · Otto-von-Guericke-Universität Magdeburg

   Affiliation as printed

   Chair of Tissue Engineering and Regenerative Medicine Wuerzburg University Hospital Wuerzburg Germany

   Institute of Molecular Biology University of Innsbruck Innsbruck Austria

   Research Center Dynamic Systems: Systems Engineering Otto‐von‐Guericke‐University Magdeburg Germany

   Chair of Tissue Engineering and Regenerative Medicine, Wuerzburg University Hospital, Wuerzburg, Germany

   Institute of Molecular Biology, University of Innsbruck, Innsbruck, Austria

   Research Center Dynamic Systems: Systems Engineering, Otto-von-Guericke-University, Magdeburg, Germany

7. Marcus Leistner corresponding

   Universitätsmedizin Göttingen · Universitätsklinikum Würzburg · University of Göttingen · Otto-von-Guericke-Universität Magdeburg

   Affiliation as printed

   Chair of Tissue Engineering and Regenerative Medicine Wuerzburg University Hospital Wuerzburg Germany

   Department of Thoracic, Cardiac and Vascular Surgery University Medical Center Goettingen Goettingen Germany

   Research Center Dynamic Systems: Systems Engineering Otto‐von‐Guericke‐University Magdeburg Germany

   Chair of Tissue Engineering and Regenerative Medicine, Wuerzburg University Hospital, Wuerzburg, Germany

   Department of Thoracic, Cardiac and Vascular Surgery, University Medical Center Goettingen, Goettingen, Germany

   Research Center Dynamic Systems: Systems Engineering, Otto-von-Guericke-University, Magdeburg, Germany