Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets.
Addressing the problem of vascularization is of vital importance when engineering three-dimensional (3D) tissues. Endothelial cells are increasingly used in tissue-engineered constructs to obtain prevascularization and to enhance in vivo neovascularization. Rat bone marrow stromal cells were cultured in thermoresponsive dishes under osteogenic conditions with human umbilical vein endothelial cells (HUVECs) to obtain homotypic or heterotypic cell sheets (CSs). Cells were retrieved as sheets from the dishes after incubation at 20 °C. Monoculture osteogenic CSs were stacked on top of homotypic or heterotypic CSs, and subcutaneously implanted in the dorsal flap of nude mice for 7 days. The implants showed mineralized tissue formation under both conditions. Transplanted osteogenic cells were found at the new tissue site, demonstrating CS bone-inductive effect. Perfused vessels, positive for human CD31, confirmed the contribution of HUVECs for the neovascularization of coculture CS constructs. Furthermore, calcium quantification and expression of osteocalcin and osterix genes were higher for the CS constructs, with HUVECs demonstrating the more robust osteogenic potential of these constructs. This work demonstrates the potential of using endothelial cells, combined with osteogenic CSs, to increase the in vivo vascularization of CS-based 3D constructs for bone tissue engineering purposes.
1] Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan [2] 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Taipas, Guimarães, Portugal [3] ICVS/3Bs PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Rogério P
RP
Pirraco
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan.
Takanori
T
Iwata
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan.
Toshiyuki
T
Yoshida
1] 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Taipas, Guimarães, Portugal [2] ICVS/3Bs PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Alexandra P
AP
Marques
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan.
Masayuki
M
Yamato
1] 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Taipas, Guimarães, Portugal [2] ICVS/3Bs PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Rui L
RL
Reis
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan.
Teruo
T
Okano
D000818:Animals; D001854:Bone Marrow Cells; D017690:Cell Transplantation; D018920:Coculture Techniques / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology*; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells; D008297:Male; D059630:Mesenchymal Stem Cells; D051379:Mice; D008819:Mice, Nude; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis / Q000502:physiology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
D019408:Platelet Endothelial Cell Adhesion Molecule-1