title: The effect of human osteoblasts on proliferation and neo-vessel formation of human umbilical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds.
abstract: Angiogenesis is a key element in early wound healing and is considered important for tissue regeneration and for directing inflammatory cells to the wound site. The improvement of vascularization by implementation of endothelial cells or angiogenic growth factors may represent a key solution for engineering bone constructs of large size. In this study, we describe a long-term culture environment that supports the survival, proliferation, and in vitro vasculogenesis of human umbilical vein endothelial cells (HUVEC). This condition can be achieved in a co-culture model of HUVEC and primary human osteoblasts (hOB) employing polyurethane scaffolds and platelet-rich plasma in a static microenvironment. We clearly show that hOB support cell proliferation and spontaneous formation of multiple tube-like structures by HUVEC that were positive for the endothelial cell markers CD31 and vWF. In contrast, in a monoculture, most HUVEC neither proliferated nor formed any apparent vessel-like structures. Immunohistochemistry and quantitative PCR analyses of gene expression revealed that cell differentiation of hOB and HUVEC was stable in long-term co-culture. The three-dimensional, FCS-free co-culture system could provide a new basis for the development of complex tissue engineered constructs with a high regeneration and vascularization capacity.
authors.affiliation: Department of Trauma Surgery, Johannes Gutenberg University School of Medicine, Langenbeckstrasse 1, D-55101 Mainz, Germany. hofmann.trauma-surgery@gmx.net
authors.forename: Alexander
authors.initials: A
authors.lastname: Hofmann
authors.forename: Ulrike
authors.initials: U
authors.lastname: Ritz
authors.forename: Sophie
authors.initials: S
authors.lastname: Verrier
authors.forename: David
authors.initials: D
authors.lastname: Eglin
authors.forename: Mauro
authors.initials: M
authors.lastname: Alini
authors.forename: Sabine
authors.initials: S
authors.lastname: Fuchs
authors.forename: C James
authors.initials: CJ
authors.lastname: Kirkpatrick
authors.forename: Pol Maria
authors.initials: PM
authors.lastname: Rommens
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006801:Humans; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic; D010006:Osteoblasts / Q000166:cytology*; D010641:Phenotype; D011140:Polyurethanes / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D054457:Tissue Scaffolds; D014471:Umbilical Veins / Q000166:cytology*
chemical_list: D015415:Biomarkers; D011140:Polyurethanes