title: A novel culture device for the evaluation of three-dimensional extracellular matrix materials.
abstract: Cell-matrix interactions in a three-dimensional (3D) extracellular matrix (ECM) are of fundamental importance in living tissue, and their in vitro reconstruction in bioartificial structures represents a core target of contemporary tissue engineering concepts. For a detailed analysis of cell-matrix interaction under highly controlled conditions, we developed a novel ECM evaluation culture device (EECD) that allows for a precisely defined surface-seeding of 3D ECM scaffolds, irrespective of their natural geometry. The effectiveness of EECD was evaluated in the context of heart valve tissue engineering. Detergent decellularized pulmonary cusps were mounted in EECD and seeded with endothelial cells (ECs) to study EC adhesion, morphology and function on a 3D ECM after 3, 24, 48 and 96 h. Standard EC monolayers served as controls. Exclusive top-surface-seeding of 3D ECM by viable ECs was demonstrated by laser scanning microscopy (LSM), resulting in a confluent re-endothelialization of the ECM after 96 h. Cell viability and protein expression, as demonstrated by MTS assay and western blot analysis (endothelial nitric oxide synthase, von Willebrand factor), were preserved at maintained levels over time. In conclusion, EECD proves as a highly effective system for a controlled repopulation and in vitro analysis of cell-ECM interactions in 3D ECM.
authors: Department of Cardiovascular Surgery, Düsseldorf University Hospital, Germany; Institute for Experimental Surgery, Düsseldorf University Hospital, Germany; Department of Cardiac Surgery, University Hospital Heidelberg, Germany.
authors: Payam
authors: P
authors: Akhyari
authors: Heiko
authors: H
authors: Ziegler
authors: Patricia
authors: P
authors: Gwanmesia
authors: Mareike
authors: M
authors: Barth
authors: Soeren
authors: S
authors: Schilp
authors: Joern
authors: J
authors: Huelsmann
authors: Stefanie
authors: S
authors: Hoffmann
authors: Julia
authors: J
authors: Bosch
authors: Gesine
authors: G
authors: Kögler
authors: Artur
authors: A
authors: Lichtenberg
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D004727:Endothelium / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D012756:Sheep; D034421:Sus scrofa