Abstract
Several studies focusing on bone tissue engineering demonstrated that given microstructuring of an implant surface has a strong effect on its interaction with cells, and their adhesion and differentiation. In the present study, geometrically structured titanium alloy surfaces are shown to be able to guide cell adhesion during differentiation in vitro. For this reason, using an electron beam texturing technique, TiAl6V4 surfaces were selectively targeted in the micrometer range. The effect of such textured titanium alloy surfaces on cell adhesion during osteogenic differentiation was analyzed for human mesenchymal stem cells (MSC), the natural precursor cells of bone tissue. Cytotoxicity, cell viability and differentiation were analyzed. Immunofluorescence stainings demonstrated that in contrast to MSC in an expansion medium, MSC in an osteogenic induction medium produce adhesion proteins such as ß3-integrins and thereby connect in an oriented way to the generated microstructures on titanium alloy surfaces. These results are of relevance for developing tailored titanium alloy implant surfaces which exhibit an improved cell response.
Acknowledgments
We thank the Department of Orthopaedic Surgery, University Clinic Aachen for providing bone spongiosa and Stephanie Brosig and Norina Labude, Institute of Pathology, RWTH Aachen University for isolation and expansion of human mesenchymal stem cells. We also thank Dr. Lüscher-Firzlaff, Institute of Biochemistry and Molecular Biology, RWTH Aachen University for the supply of PCR equipment and Manfred Bovi, Facility of Electron Microscopy, RWTH Aachen University Hospital for his help in the SEM analysis.
References
[1] Anselme K. Osteoblast adhesion on biomaterials. Biomaterials 2000; 21: 667–681.10.1016/S0142-9612(99)00242-2Search in Google Scholar
[2] Ball M, Grant DM, Lo WJ, Scotchford CA. The effect of different surface morphology and roughness on osteoblast-like cells. J Biomed Mater Res A 2008; 86: 637–647.10.1002/jbm.a.31652Search in Google Scholar
[3] Brammer KS, Choi C, Frandsen CJ, Oh S, Jin S. Hydrophobic nanopillars initiate mesenchymal stem cell aggregation and osteo-differentiation. Acta Biomater 2011; 7: 683–690.10.1016/j.actbio.2010.09.022Search in Google Scholar
[4] Brunette DM. Spreading and orientation of epithelial cells on grooved substrata. Exp Cell Res 1986; 167: 203–217.10.1016/0014-4827(86)90217-XSearch in Google Scholar
[5] Curtis AS, Gadegaard N, Dalby MJ, Riehle MO, Wilkinson CD, Aitchison G. Cells react to nanoscale order and symmetry in their surroundings. IEEE Trans Nanobiosci 2004; 3: 61–65.10.1109/TNB.2004.824276Search in Google Scholar
[6] Dalby MJ, Gadegaard N, Curtis AS, Oreffo RO. Nanotopographical control of human osteoprogenitor differentiation. Curr Stem Cell Res Ther 2007; 2: 129–138.10.2174/157488807780599220Search in Google Scholar
[7] Dalby MJ, Riehle MO, Johnstone H, Affrossman S, Curtis AS. Investigating the limits of filopodial sensing: a brief report using SEM to image the interaction between 10 nm high nano-topography and fibroblast filopodia. Cell Biol Int 2004; 28: 229–236.10.1016/j.cellbi.2003.12.004Search in Google Scholar
[8] Gittens R, McLachlan T, Olivares-Navarrete R, et al. The effects of combined micron-submicron-scale surface roughness and nanoscale features on cell proliferation and differentiation. Biomaterials 2011; 32: 3395–3403.10.1016/j.biomaterials.2011.01.029Search in Google Scholar
[9] Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 2005; 26: 5474–5491.10.1016/j.biomaterials.2005.02.002Search in Google Scholar
[10] Kumar A, Bhat V, Balakrishnan M, et al. Bioactivity and surface characteristics of titanium implants following various surface treatments: an in vitro study. J Oral Implant 2014; [Epub ahead of print].Search in Google Scholar
[11] Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone. Nature 2014; 507: 323–328.10.1038/nature13145Search in Google Scholar
[12] Le Blank K, Pittenger M. Mesenchymal stem cells: progress toward promise. Cytotherapy 2005; 7: 36–45.10.1016/S1465-3249(05)70787-8Search in Google Scholar
[13] Lehnert D, Wehrle-Haller B, David C, et al. Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion. J Cell Sci 2004; 117: 41–52.10.1242/jcs.00836Search in Google Scholar
[14] Lenhert S, Meier MB, Meyer U, Chi L, Wiesmann HP. Osteoblast alignment, elongation and migration on grooved polystyrene surfaces patterned by Langmuir-Blodgett lithography. Biomaterials 2005; 26: 563–570.10.1016/j.biomaterials.2004.02.068Search in Google Scholar
[15] Lim YW, Kwon SY, Sun DH, Kim HE, Kim YS. Enhanced cell integration to titanium alloy by surface treatment with microarc oxidation: a pilot study. Clin Orthop Relat Res 2009; 467: 2251–2258.10.1007/s11999-009-0879-6Search in Google Scholar
[16] Linder L, Carlsson A, Marsal L, Bjursten LM, Branemark PI. Clinical aspects of osseointegration in joint replacement. J Bone Joint Surg Br 1988; 70: 550–555.10.1302/0301-620X.70B4.3403596Search in Google Scholar
[17] Lüthen F, Lange R, Becker P, Rychly J, Beck U, Nebe JG. The influence of surface roughness of titanium on ß1- and ß3-integrin adhesion and the organization of fibronectin in human osteoblastic cells. Biomaterials 2005; 26: 2423–2440.10.1016/j.biomaterials.2004.07.054Search in Google Scholar
[18] Monsees TK, Barth K, Tippelt S, et al. Effects of different titanium alloys and nanosize surface patterning on adhesion, differentiation, and orientation of osteoblast-like cells. Cells Tissues Organs 2005; 180: 81–95.10.1159/000086749Search in Google Scholar
[19] Neuss S, Apel C, Buttler P, et al. Assessment of stem cell/biomaterial combinations for stem cell-based tissue engineering. Biomaterials 2008; 29: 302–313.10.1016/j.biomaterials.2007.09.022Search in Google Scholar
[20] Neuss S, Blomenkamp I, Stainforth R, et al. The use of a shape-memory poly(ε-caprolactone) dimethacrylate network as a tissue engineering scaffold. Biomaterials 2009; 30: 1697–1705.10.1016/j.biomaterials.2008.12.027Search in Google Scholar
[21] Neuss S, Stainforth R, Salber J, et al. Long-term survival and bipotent terminal differentiation of human mesenchymal stem cells (hMSC) in combination with a commercially available three-dimensional collagen scaffold. Cell Transplant 2008; 17: 977–986.10.3727/096368908786576462Search in Google Scholar
[22] Oh S, Brammer KS, Li YS, et al. Stem cell fate dictated solely by altered nanotube dimension. Proc Natl Acad Sci USA 2009; 106: 2130–2135.10.1073/pnas.0813200106Search in Google Scholar
[23] Olivares-Navarrete R, Hyzy SL, Pan Q, et al. Osteoblast maturation on microtextured titanium involves paracrine regulation of bone morphogenic protein signaling. J Biomed Mater Res A 2014; doi: 10.1002/jbm.a.35308 Epub ahead of print.10.1002/jbm.a.35308Search in Google Scholar
[24] Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143–147.10.1126/science.284.5411.143Search in Google Scholar
[25] Puckett S, Pareta R, Webster TJ. Nano rough micron patterned titanium for directing osteoblast morphology and adhesion. Int J Nanomedicine 2008; 3: 229–241.Search in Google Scholar
[26] Puleo DA, Holleran LA, Doremus RH, Bizios R. Osteoblast responses to orthopedic implant materials in vitro. J Biomed Mater Res 1991; 25: 711–723.10.1002/jbm.820250603Search in Google Scholar
[27] Rice JM, Hunt JA, Gallagher JA, Hanarp P, Sutherland DS, Gold J. Quantitative assessment of the response of primary derived human osteoblasts and macrophages to a range of nanotopography surfaces in a single culture model in vitro. Biomaterials 2003; 24: 4799–4818.10.1016/S0142-9612(03)00381-8Search in Google Scholar
[28] Santander S, Alcaine C, Lyahyai J, et al. In vitro osteoinduction of human mesenchymal stem cells in biomimetic surface modified titanium alloy implants. Dent Mater J 2014; 33: 305–312.10.4012/dmj.2012-015-rSearch in Google Scholar
[29] Schneider RK, Neuss S, Knüchel R, Perez-Bouza A. Mesenchymal stem cells for bone tissue engineering. Pathologe 2010; 31: 138–146.10.1007/s00292-010-1329-7Search in Google Scholar
[30] Schneider RK, Puellen A, Kramann R, et al. The osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. Biomaterials 2010; 31: 467–480.10.1016/j.biomaterials.2009.09.059Search in Google Scholar
[31] Schultz H. Elektronenstrahlschweißen. 2nd ed. Düsseldorf: DVS-Verlag 2000.Search in Google Scholar
[32] Sumanasinghe RD, Osborne JA, Loboa EG. Mesenchymal stem cell-seeded collagen matrices for bone repair: effects of cyclic tensile strain, cell density, and media conditions on matrix contraction in vitro. J Biomed Mater Res A 2009; 88: 778–786.10.1002/jbm.a.31913Search in Google Scholar
[33] Thomas G, Vincent R, Matthews G, Dance B, Grant PS. Interface topography and residual stress distributions in coatings for fusion armour applications. Mat Sci Eng A Struct 2008; 477: 35–42.10.1016/j.msea.2007.05.120Search in Google Scholar
[34] Wang JH, Grood ES, Florer J, Wenstrup R. Alignment and proliferation of MC3T3-E1 osteoblasts in microgrooved silicone substrata subjected to cyclic stretching. J Biomech 2000; 33: 729–735.10.1016/S0021-9290(00)00013-0Search in Google Scholar
[35] Wang JH, Liu YZ, Yin LJ, et al. BMP9 and COX-2 form an important regulatory loop in BMP9-induced osteogenic differentiation of mesenchymal stem cells. Bone 2013; 57: 311–321.10.1016/j.bone.2013.08.015Search in Google Scholar
[36] Weiss P. Interaction between cells. Rev Mod Phys 1959; 31: 11–20.10.1103/RevModPhys.31.11Search in Google Scholar
[37] Yang J, Wang J, Yuan T, et al. The enhanced effect of surface microstructured porous titanium on adhesion and osteoblastic differentiation of mesenchymal stem cells. J Mater Sci Mater Med 2013; 24: 2235–2246.10.1007/s10856-013-4976-4Search in Google Scholar
[38] Zhang W, Li Z, Huang Q, et al. Effects of a hybrid micro/nanorod topography-modified titanium implant on adhesion and osteogenic differentiation in rat bone marrow mesenchymal stem cells. Int J Nanomedicine 2013; 8: 257–265.Search in Google Scholar
[39] Zhao G, Zinger O, Schwartz Z, Wieland M, Landolt D, Boyan BD. Osteoblast-like cells are sensitive to submicron-scale surface structure. Clin Oral Implant Res 2006; 17: 258–264.10.1111/j.1600-0501.2005.01195.xSearch in Google Scholar
©2015 by De Gruyter