Abstract
Osteogenesis requires close co-operation with angiogenesis to create vascularized bone tissue. In this study, an indirect co-culture model using osteoblasts (OBs), primary endothelial cells (ECs) and Matrigel interlayer was established to understand the impact of each cell type on the other. ECs synergistically enhanced osteoblastic gene expression by OBs, while OBs were capable of supporting tubule-like structures formed by ECs on Matrigel, enhancing mean tubule length from 146.5 ± 23.5 μm in ECs alone to 192 ± 28.6 μm in co-culture (p < 0.05). Similar improvements were noted in terms of tubule number. An applicability study of the co-culture model to bone tissue engineering, performed on a biopolymer fibrous membrane, showed substantially enhanced deposition of calcified nodules. These results demonstrate the efficacy of co-culture with ECs to improve osteogenesis for bone tissue engineering.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Buser D, Schenk RK, Steinemann S, Fiorellini JP, Fox CH, Stich H (1991) Influence of surface characteristics on bone integration of titanium implants. A histomorphometric study in miniature pigs. J Biomed Mater Res 25:889–902
Donos N, Retzepi M, Wall I, Hamlet S, Ivanovski S (2011) In vivo gene expression profile of guided bone regeneration associated with a microrough titanium surface. Clin Oral Implants Res 22:390–398
Fujio M, Yamamoto A, Ando Y, Shohara R, Kinoshita K, Kaneko T, Hibi H, Ueda M (2011) Stromal cell-derived factor-1 enhances distraction osteogenesis-mediated skeletal tissue regeneration through the recruitment of endothelial precursors. Bone 49:693–700
Gerstenfeld LC, Cullinane DM, Barnes GL, Graves DT, Einhorn TA (2003) Fracture healing as a post-natal developmental process: molecular, spatial, and temporal aspects of its regulation. J Cell Biochem 88:873–884
Gotfredsen K, Wennerberg A, Johansson C, Skovgaard LT, Hjorting-Hansen E (1995) Anchorage of TiO2-blasted, HA-coated, and machined implants: an experimental study with rabbits. J Biomed Mater Res 29:1223–1231
Grellier M, Granja PL, Fricain JC, Bidarra SJ, Renard M, Bareille R, Bourget C, Amedee J, Barbosa MA (2009) The effect of the co-immobilization of human osteoprogenitors and endothelial cells within alginate microspheres on mineralization in a bone defect. Biomaterials 30:3271–3278
Hofmann A, Ritz U, Verrier S, Eglin D, Alini M, Fuchs S, Kirkpatrick CJ, Rommens PM (2008) 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. Biomaterials 29:4217–4226
Kim SS, Park MS, Cho SW, Kang SW, Ahn KM, Lee JH, Kim BS (2010) Enhanced bone formation by marrow-derived endothelial and osteogenic cell transplantation. J Biomed Mater Res A 92:246–253
Luo T, Zhang W, Shi B, Cheng X, Zhang Y (2012) Enhanced bone regeneration around dental implant with bone morphogenetic protein 2 gene and vascular endothelial growth factor protein delivery. Clin Oral Implants Res 23:467–473
Maes C, Carmeliet G, Schipani E (2012) Hypoxia-driven pathways in bone development, regeneration and disease. Nat Rev Rheumatol 8(6):358–366
Matsubara H, Hogan DE, Morgan EF, Mortolock DP, Einhorn TA, Gerstenfeld LC (2012) Vascular tissues are a primary source of BMP2 expression during bone formation induced by distraction osteogenesis. Bone 51(1):168–180
Samee M, Kasugai S, Kondo H, Ohya K, Shimokawa H, Kuroda S (2008) Bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) transfection to human periosteal cells enhances osteoblast differentiation and bone formation. J Pharmacol Sci 108:18–31
Shah AR, Shah SR, Oh S, Ong JL, Wenke JC, Agrawal CM (2011) Migration of co-cultured endothelial cells and osteoblasts in composite hydroxyapatite/polylactic acid scaffolds. Ann Biomed Eng 39:2501–2509
Song X, Liu S, Qu X, Hu Y, Zhang X, Wang T, Wei F (2011) BMP2 and VEGF promote angiogenesis but retard terminal differentiation of osteoblasts in bone regeneration by up-regulating Id1. Acta Biochim Biophys Sin (Shanghai) 43:796–804
Steinbrech DS, Mehrara BJ, Saadeh PB, Chin G, Dudziak ME, Gerrets RP, Gittes GK, Longaker MT (1999) Hypoxia regulates VEGF expression and cellular proliferation by osteoblasts in vitro. Plastic Reconst Sur 104:738–747
Unger RE, Sartoris A, Peters K, Motta A, Migliaresi C, Kunkel M, Bulnheim U, Rychly J, Kirkpatrick CJ (2007) Tissue-like self-assembly in cocultures of endothelial cells and osteoblasts and the formation of microcapillary-like structures on three-dimensional porous biomaterials. Biomaterials 28:3965–3976
Zelzer E, McLean W, Ng YS, Fukai N, Reginato AM, Lovejoy S, D’Amore PA, Olsen BR (2002) Skeletal defects in VEGF (120/120) mice reveal multiple roles for VEGF in skeletogenesis. Development 129:1893–1904
Acknowledgments
This study was supported by grants from Priority Research Centers Program (2009-0093829) and WCU program (R31-2008-000-100069-0), National Research Foundation, South Korea.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Tuvd Dariima and Guang-Zhen Jin contributed equally to this study.
Rights and permissions
About this article
Cite this article
Dariima, T., Jin, GZ., Lee, EJ. et al. Cooperation between osteoblastic cells and endothelial cells enhances their phenotypic responses and improves osteoblast function. Biotechnol Lett 35, 1135–1143 (2013). https://doi.org/10.1007/s10529-013-1170-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-013-1170-1