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In vivo engineering of a human vasculature for bone tissue engineering applications.

Steffens L, Wenger A, Stark GB, Finkenzeller G - J. Cell. Mol. Med. (2009)

Bottom Line: In both assays, the development of a complex three-dimensional network of perfused human neovessels could be detected.After subcutaneous implantation into immunodeficient mice, the newly formed human vasculature was stabilized by the recruitment of murine smooth muscle alpha-actin-positive mural cells and anastomoses with the mouse vasculature.We conclude that this endothelial cell spheroid system can be used to create a network of functional perfused blood vessels in vivo.

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Affiliation: Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Freiburg, Germany.

ABSTRACT
The neovascularization of three-dimensional voluminous tissues, such as bone, represents an important challenge in tissue engineering applications. The formation of a preformed vascular plexus could maintain cell viability and promote vascularization after transplantation. We have developed a three-dimensional spheroidal coculture system consisting of human primary endothelial cells and human primary osteoblasts (hOBs) to improve angiogenesis in bone tissue engineering applications. In this study, we investigated the survival and vascularization of the engineered implants in vivo. Endothelial cell spheroids were cocultured with hOBs in fibrin and seeded into scaffolds consisting of processed bovine cancellous bone (PBCB). The cell-seeded scaffolds were evaluated for their angiogenic potential in two different in vivo assays: the chick embryo chorioallantoic membrane (CAM) model and the severe combined immunodeficiency disorder (SCID) mouse model. In both assays, the development of a complex three-dimensional network of perfused human neovessels could be detected. After subcutaneous implantation into immunodeficient mice, the newly formed human vasculature was stabilized by the recruitment of murine smooth muscle alpha-actin-positive mural cells and anastomoses with the mouse vasculature. We conclude that this endothelial cell spheroid system can be used to create a network of functional perfused blood vessels in vivo. The finding that this process takes place with high efficacy in the presence of co-implanted primary osteoblasts and in an osteoconductive environment provided by the PBCB scaffold, suggests that this system may be suitable for improving vascularization in bone tissue engineering.

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Macroscopic image of the osteoconductive PBCB scaffold.
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fig01: Macroscopic image of the osteoconductive PBCB scaffold.

Mentions: Discs of PBCB (Fig. 1) were seeded with fibrin gel-immobilized hOBs and HUVEC-spheroids. After 24 hrs in vitro, cell-seeded scaffolds were inspected by light microscopy revealing integrity of the spherical cell cluster configuration (Fig. 2A).


In vivo engineering of a human vasculature for bone tissue engineering applications.

Steffens L, Wenger A, Stark GB, Finkenzeller G - J. Cell. Mol. Med. (2009)

Macroscopic image of the osteoconductive PBCB scaffold.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4516493&req=5

fig01: Macroscopic image of the osteoconductive PBCB scaffold.
Mentions: Discs of PBCB (Fig. 1) were seeded with fibrin gel-immobilized hOBs and HUVEC-spheroids. After 24 hrs in vitro, cell-seeded scaffolds were inspected by light microscopy revealing integrity of the spherical cell cluster configuration (Fig. 2A).

Bottom Line: In both assays, the development of a complex three-dimensional network of perfused human neovessels could be detected.After subcutaneous implantation into immunodeficient mice, the newly formed human vasculature was stabilized by the recruitment of murine smooth muscle alpha-actin-positive mural cells and anastomoses with the mouse vasculature.We conclude that this endothelial cell spheroid system can be used to create a network of functional perfused blood vessels in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Freiburg, Germany.

ABSTRACT
The neovascularization of three-dimensional voluminous tissues, such as bone, represents an important challenge in tissue engineering applications. The formation of a preformed vascular plexus could maintain cell viability and promote vascularization after transplantation. We have developed a three-dimensional spheroidal coculture system consisting of human primary endothelial cells and human primary osteoblasts (hOBs) to improve angiogenesis in bone tissue engineering applications. In this study, we investigated the survival and vascularization of the engineered implants in vivo. Endothelial cell spheroids were cocultured with hOBs in fibrin and seeded into scaffolds consisting of processed bovine cancellous bone (PBCB). The cell-seeded scaffolds were evaluated for their angiogenic potential in two different in vivo assays: the chick embryo chorioallantoic membrane (CAM) model and the severe combined immunodeficiency disorder (SCID) mouse model. In both assays, the development of a complex three-dimensional network of perfused human neovessels could be detected. After subcutaneous implantation into immunodeficient mice, the newly formed human vasculature was stabilized by the recruitment of murine smooth muscle alpha-actin-positive mural cells and anastomoses with the mouse vasculature. We conclude that this endothelial cell spheroid system can be used to create a network of functional perfused blood vessels in vivo. The finding that this process takes place with high efficacy in the presence of co-implanted primary osteoblasts and in an osteoconductive environment provided by the PBCB scaffold, suggests that this system may be suitable for improving vascularization in bone tissue engineering.

Show MeSH
Related in: MedlinePlus