Tissue engineered scaffolds must have an organized and repeatable microstructure which enables cells to assemble in an ordered matrix that allows adequate nutriental perfusion. In this work, to evaluate the reciprocal cell interactions of endothelial and osteoblast-like cells, human osteoblast-like cells (MG63) and Human Umbilical Vein Endothelial Cells (HUVEC) were co-seeded onto 3D geometrically controlled porous poly(ε-caprolactone) (PCL) and cultured by means of a rotary cell culture system (RCCS-4DQ). In our dynamic co-culture system, the lack of significant enhancement of osteoblast ALP activity and ECM production indicated that the microgravity conditions of the rotary system affected the cells by favoring their proliferation and cellular cross-talk. These results emphasize how osteoblasts increase endothelial cell proliferate and endothelial cells amplify the growth of osteoblasts but decrease their differentiation. This dynamic seeding of osteoblasts and endothelial cells onto a 3D polymeric scaffold may represent a unique approach for studying the mechanisms of interaction of endothelial and osteoblast cells as well as achieve a functional hybrid in which angiogenesis, furnished by neo-vascular organization of endothelial cells may further support osteoblasts growth. Furthermore, this in vitro model may be useful in examining the applicability of novel material structures for tissue engineering.
Hutmacher, D.W. (2001). Scaffold Design and Fabrication Technologies for Engineering Tissues-State of the Art and Future Perspectives , J. Biomater. Sci. Polym. Ed., 12: 107-24.
2.
Cima, L.G., Vacanti, J.P., Vacanti, C., Ingber, D., Mooney, D. and Langer, R. (1991). Tissue Engineering by Cell Transplantation Using Degradable Polymer Substrates, J. Biomech. Eng., 113: 143-151.
Ciapetti, G. , Ambrosio, L., Savarino , L., Granchi, D., Cenni, E., Baldini, N., et al. (2003 ). Osteoblast Growth and Function in Porous Poly e-caprolactone Matrices for Bone Repair: A Preliminary Study, Biomaterials , 24: 3815-3824.
5.
Williams, J.M. , Adewunmi, A., Schek , R.M., Flanagan, C.L., Krebsbach, P.H., Feinberg, S.E., et al. ( 2005). Bone Tissue Engineering Using Polycaprolactone Scaffolds Fabricated Via Selective Laser Sintering, Biomaterials, 26: 4817-4827.
6.
Li, W., Tuli, R., Huang, X., Laquerriere, P. and Tuan, R.S. (2005). Multilineage Differentiation of Human Mesenchymal Stem Cells in a Three-dimensional Nanofibrous Scaffold, Biomaterials , 26: 5158-5166.
7.
Unger, R.E., Sartoris, A., Peters, K., Motta, A., Migliaresi, C., Kunkel, M., et al. (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.
8.
Decker, B., Bartles, H. and Decker, S. (1995). Relationships Between Endothelial Cells, Pericytes, and Osteoblasts During Bone Formation in the Sheep Femur Following Implantation of Tricalciumphosphate-ceramic, Anat. Rec., 242: 310-320.
9.
Villars, F., Bordenave, L., Bareille, R. and Amedee, J. (2000). Effect of Human Endothelial Cells on Human Bone Marrow Stromal Cell Phenotype: Role of VEGF?J. Cell. Biochem. , 79: 679-685.
10.
Carrington, J.L. and Reddi, A.H. (1991). Parallels Between Development of Embryonic and Matrix-Induced Endochondral Bone, Bioassay , 13: 403-408.
11.
Collin-Osdoby, P. (1994). Role of Vascular Endothelial Cells in Bone Biology, J. Cell. Biochem., 55: 304-309.
12.
Wang, D.S., Miura, M., Demura, H. and Sato, K. (1997). Anabolic Effects of 1,25-dihydroxyvitamin D3 on Osteoblasts are Enhanced by Vascular Endothelial Growth Factor Produced by Osteoblasts and by Growth Factors Produced by Endothelial Cells, Endocrinology, 138: 2953-2962.
13.
Botchwey, E.A. , Dupree, M.A., Pollack , S.R., Levine, E.M. and Laurencin , C.T. (2003). Tissue Engineered Bone: Measurement of Nutrient Transport in Three-dimensional Matrices, J. Biomed. Mater. Res. A., 67: 357-367.
14.
Karande, T.S. , Ong, J.L. and Agrawal, C.M. (2004). Diffusion in Musculoskeletal Tissue Engineering Scaffolds: Design Issues Related to Porosity, Permeability, Architecture, and Nutrient Mixing, Ann. Biomed. Eng., 32: 1728-1743.
15.
Anselme, K. (2000). Osteoblast Adhesion to Materials, Biomaterials, 21: 667-681.
16.
Torricelli, P., Fini, M., Giavaresi , G., Borsari, V., Carpi, A., Nicolini, A., et al. (2003 ). Comparative Interspecies Investigation on Osteoblast Cultures: Data on Cell Viability and Synthetic Activity, Biomed. Pharmacother. , 57(1): 57-62.
17.
Gibson, L.J. and Ashby, M.F. (1997). Cellular Solids: Structure and Properties, 2nd edn, Cambridge University Press, Cambridge, UK.
18.
Wenger, A., Stahl, A., Weber, H., Finkenzeller, G., Augustin, H.G., Stark, G.B. and Kneser, U. (2004). Modulation of in vitro Angiogenesis in a Three-dimensional Spheroidal Coculture Model for Bone Tissue Engineering, Tissue Eng., 10: 1536-1547.
19.
Unger, R.E., Krump-Konvalinkova, V., Peters, K. and Kirkpatrick, C.J. (2002). In vitro Expression of the Endothelial Phenotype: Comparative study of Primary Isolated Cells and Cell Lines, Including the Novel Cell Line HPMEC-ST1.6R, Microvasc. Res., 64: 384-397.
20.
Muller, A.M. , Hermanns, M.I., Skrzynski, C., Nesslinger, M., Muller, K.M. and Kirkpatrick , C.J. (2002). Expression of the Endothelial Markers PECAM-1, vWf, and CD34 in vivo and in vitro, Exp. Mol. Pathol., 72: 221-229.
21.
Hyukjin, L., Hyun, J.C. and Tae, G.P. (2007). Perspectives On: Local and Sustained Delivery of Angiogenic Growth Factors, J. Bioactive. Comp. Polymers , 22: 89-114.
22.
Konstantinov, S.M., Mindova, M.M., Gospodinov , P.T. and Genova, P.I. (2004). Three-Dimensional Bioreactor Cultures: A Useful Dynamic Model for the Study of Cellular Interactions, Ann. NY Acad. Sci., 1030: 103-115.
23.
Ingber, D.E. (1997). Integrins, Tensegrity, and Mechanotransduction, Gravit. Space Biol. Bull., 10: 49-55.
24.
Ingber, D.E. (1999). How Cells (might) Sense Microgravity, FASEB J., 13: S3-15.
25.
Ingber, D.E. (2002). Mechanical Signaling and the Cellular Response to Extracellular Matrix in Angiogenesis and Cardiovascular Physiology, Circ. Res., 91: 877-887.
26.
Carlsson, S.I. , Bertilaccio, M.T., Ballabio , E. and Maier, J.A. (2003). Endothelial Stress by Gravitational Unloading: Effects on Cell Growth and Cytoskeletal Organization, Biochim. Biophys. Acta, 1642: 173-179.
27.
Carmeliet, G. and Bouillon, R. (1999). The Effect of Microgravity on Morphology and Gene Expression of Osteoblasts in vitro , FASEB J., 13: S129-S134.
28.
Jones, A.R. , Clark, C.C. and Brighton, C.T. (1995). Microvessel Endothelial Cells and Pericytes Increase Proliferation and Repress Osteoblast Phenotypic Markers in Rat Calvarial Bone Cell Cultures, J. Orthop. Res. , 13: 553-561.
29.
Villars, F. , Guillotin, B., Amedee , T., Dutoya, S., Bordenave, L., Bareille, R., et al. (2002 ). Effect of HUVEC on Human Osteoprogenitor Cell Differentiation Needs Heterotypic Gap Junction Communication, Am. J. Physiol. Cell Physiol., 282: C775-85.
30.
Mattioli-Belmonte, M., Lucarini, G., Virgili , C.L., Biagini, G., Detomaso, L., Favia, P., et al. (2005). Mesenchymal Stem Cells on Plasma-Deposited Acrylic Acid Coatings: An In Vitro Investigation to Improve Biomaterial Performance in Bone Reconstruction, J. Bioactive Comp. Polymers, 20: 343-360.
