title: Two- and three-dimensional co-culture models of soft tissue healing: pericyte-endothelial cell interaction.
abstract: The demographic change in western countries towards an older population is being shadowed by an increased appearance of chronic diseases influencing soft tissue healing in a negative manner. Although various promising therapeutic approaches are available for treating chronic wounds, no in vitro model exists that successfully allows the analysis of interacting cells and of the effect of therapeutic drugs within a wound. Granulation tissue assures wound stability, neo-angiogenesis and revascularization finally leading to functional soft tissue repair. As one of the first steps in developing a model for human granulation tissue, we examined microvascular endothelial cells and pericytes in conventional 2D and in 3D spheroid co-cultures. We determined which parameters could be used in a standardized manner and whether the cultures were responsive to hypoxia and to erythropoietin supplementation. The read-out parameters of cell migration, cell density, rate of apoptotic cells, spatial cell distribution in the spheroid and spheroid volume were shown to be excellent analytic measures. In addition, quantification of hypoxia-related genes identified a total of 13 genes that were up-regulated in spheroids after hypoxia. As these parameters delivered reliable results in the present approach and as the general morphological distribution of pericytes and endothelial cells within the spheroid occurred in a typical manner, we believe that this basic in vitro model will serve for the future study of diverse aspects of soft tissue healing.
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Jennewein
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Monika
authors.initials: M
authors.lastname: Bubel
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Guthörl
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Metzger
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Weigert
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Pohlemann
authors.affiliation: Department of Trauma, Hand and Reconstructive Surgery, Saarland University, Kirrberger Strasse, Building 57, 66421, Homburg, Germany. martin.oberringer@uks.eu.
authors.forename: Martin
authors.initials: M
authors.lastname: Oberringer
mesh_terms: D017209:Apoptosis; D002450:Cell Communication; D002452:Cell Count; D018920:Coculture Techniques / Q000379:methods*; D020405:Dermis / Q000098:blood supply; D042783:Endothelial Cells / Q000166:cytology*; D005786:Gene Expression Regulation; D006097:Granulation Tissue / Q000378:metabolism / Q000473:pathology; D006801:Humans; D055806:Microvessels / Q000166:cytology; D008954:Models, Biological; D020286:Pericytes / Q000166:cytology*; D016133:Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D014945:Wound Healing
keywords: Granulation tissue; Hypoxia; Microvascular endothelial cells; Pericytes; Wound healing