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Vol. 180, No. 2, 2005
Issue release date: 2005

Effects of Different Titanium Alloys and Nanosize Surface Patterning on Adhesion, Differentiation, and Orientation of Osteoblast-Like Cells

Monsees T.K. · Barth K. · Tippelt S. · Heidel K. · Gorbunov A. · Pompe W. · Funk R.H.W.
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To test nanosize surface patterning for application as implant material, a suitable titanium composition has to be found first. Therefore we investigated the effect of surface chemistry on attachment and differentiation of osteoblast-like cells on pure titanium prepared by pulsed laser deposition (TiPLD) and different Ti alloys (Ti6Al4V, TiNb30 and TiNb13Zr13). Early attachment (30 min) and alkaline phosphatase (ALP) activity (day 5) was found to be fastest and highest, respectively, in cells grown on TiPLD and Ti6Al4V. Osteoblasts seeded on TiPLD produced most osteopontin (day 10), whereas expression of this extracellular matrix protein was an order of magnitude lower on the TiNb30 surface. In contrast, expression of the corresponding receptor, CD44, was not influenced by surface chemistry. Thus, TiPLD was used for further experiments to explore the influence of surface nanostructures on osteoblast adhesion, differentiation and orientation. By laser-induced oxidation, we produced patterns of parallel Ti oxide lines with different widths (0.2–10 µm) and distances (2–20 and 1,000 µm), but a common height of only 12 nm. These structures did not influence ALP activity (days 5–9), but had a positive effect on cell alignment. Two days after plating, the majority of the focal contacts were placed on the oxide lines. The portion of larger focal adhesions bridging two lines was inversely related to the line distance (2–20 µm). In contrast, the portion of aligned cells did not depend on the line distance. On average, 43% of the cells orientated parallel towards the lines, whereas 34% orientated vertically. In the control pattern (1,000 µm line distance), cell distribution was completely at random. Because a significant surplus of the cells preferred a parallel alignment, the nanosize difference in height between Ti surface and oxide lines may be sufficient to orientate the cells by contact guiding. However, gradients in electrostatic potential and surface charge density at the Ti/Ti oxide interface may additionally influence focal contact formation and cell guidance.

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  1. Bagambisa, F.B., H.F. Kappert, W. Schilli (1994) Cellular and molecular biological events at the implant interface. J Craniomaxillofac Surg 22: 12–17.
  2. Baxter, L.C., V. Frauchiger, M. Textor, I. ap Gwynn, R.G. Richards (2002) Fibroblast and osteoblast adhesion and morphology on calcium phosphate surfaces. Eur Cell Mater 4: 1–17.
  3. Becker, D., U. Geiβler, U. Hempel, S. Bierbaum, D. Scharnweber, H. Worch, K.-W. Wenzel (2002) Proliferation and differentiation of rat calvarial osteoblasts on type I collagen-coated titanium alloy. J Biomed Mater Res 59: 516–527.
  4. Bellows, C.G., J.E. Aubin, J.N. Heersche, M.E. Antosz, (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38: 143–154.
  5. Bellows, C.G., J.N. Heersche, J.E. Aubin (1999) Aluminium accelerates osteoblastic differentiation but is cytotoxic in long-term rat calvaria cell cultures. Calcif Tissue Int 65: 59–65.
  6. Boyan, B.D., R. Batzer, K. Kiesewetter, Y. Lie, D.L. Cochran, S. Szmuckler-Moncler, D.D. Dean, Z. Schwartz (1998) Titanium surface roughness alters responsiveness of MG63 osteoblastic-like cells to 1α,25-(OH)2D3. J Biomed Mater Res 39: 77–85.
  7. Breme, J., V. Biehl, A. Hoffmann (2000) Tailor-made composites based on titanium for medical devices. Adv Eng Mater 2: 270–275.
  8. Bretaudier, J. P., T. Spillman (1984) Alkaline phosphatases; in Bergmeyer, H. U. (ed): Methods of Enzymatic Analysis. Weinheim, Verlag Chemie, pp 75–86.
  9. Brunette, D.M. (1986) Spreading and orientation of epithelial cells on grooved substrata. Exp Cell Res 167: 203–217.
  10. Brunette, D.M., B. Cheroudi (1999) The effects of the surface topography of micromachined titanium substrata on cell behavior in vitro and in vivo. J Biomech Eng 121: 49–57.
  11. Burridge, K., M. Chrzanowska-Wodnicka (1996) Focal adhesions, contractility, and signaling. Annu Rev Cell Biol 12: 463–518.
  12. Carvalho, R.S., J. L. Schaffer, L.C. Gerstenfeld (1998) Osteoblasts induce osteopontin expression in response to attachment on fibronectin: demonstration of a common role for integrin receptors in the signal transduction process of cell attachment and mechanical stimulation. J Cell Biochem 70: 376–390.
  13. Clark, P., P. Connolly, A.S. Curtis, J.A. Dow, C. D. Wilkinson (1991) Cell guidance by ultrafine topography. J Cell Sci 99: 73–77.
  14. Curtis, A.S., N. Gadegaard, M.J. Dalby, M.O. Riehle, C.D. Wilkinson, G. Aitchison (2004) Cells react to nanoscale order and symmetry in their surroundings. IEEE Trans Nanobiosci 3: 61–65.
  15. Dalby, M.J., S.J. Yarwood, M.O. Riehle, H.J.H. Johnstone, S. Affrossman, A.S.G. Curtis (2002) Increasing fibroblast response to materials using nanotopography: morphological and genetic measurements of cell response to 13 nm high polymer demixed islands. Exp Cell Res 276: 1–9.
  16. Dalby, M.J., M.O. Riehle, H. Johnstone, S. Affrossman, A.S.G. Curtis (2004) 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 28: 229–236.
  17. Dalby, M.J., M.O. Riehle, D.S. Sutherland, H. Agheli, A.S.G. Curtis (2005) Morphological and microarray analysis of human fibroblasts cultured on nanocolumns produced by colloidal lithography. Eur Cell Mater 9: 1–8.
  18. Degasne, I., M.F. Basle, V. Demais, G. Hure, M. Lesourd, B. Grolleau, L. Mercier, D. Chappard (1999) Effects of roughness, fibronectin and vitronectin on attachment, spreading, and proliferation of human osteoblast-like cells (Saos-2) on titanium surfaces. Calcif Tissue Int 64: 499–507.
  19. De Santis, D., C. Guerriero, P.F. Nocini, A. Ungersbock, G. Richards, P. Gotte, U. Armato (1996) Adult human cells from jaw bones cultured on plasma-sprayed or polished surfaces of titanium or hydroxyapatite discs. J Mater Sci Mater Med 7: 21–28.
  20. Eisenbarth, E., P. Linez, V. Biehl, D. Velten, J. Breme, H. Hildebrand (2002) Cell orientation and cytoskeleton organization on ground titanium surfaces. Biomol Eng 19: 233–237.
  21. Erskine, L., C.D. McCaig (1997) Integrated interactions between chondroitin sulphate proteoglycans and weak dc electric fields regulate nerve growth cone guidance in vitro. J Cell Sci 110: 1957–1965.
  22. Ferrier, J., S.M. Ross, J. Kanehisa, J.E. Aubin (1986) Osteoclasts and osteoblasts migrate in opposite directions in response to a constant electrical field. J Cell Physiol 129: 283–288.
  23. Geissler, U., U. Hempel, C. Wolf, D. Scharnweber, H. Worch, K.W. Wenzel (2000) Collagen type I-coating of Ti6Al4V promotes adhesion of osteoblasts. J Biomed Mater Res 51: 752–760.
  24. Gerischer, H. (1989) Models for the discussion of the photo-electrochemical response of oxide layers on metals. Corr Sci 29: 257–266.
  25. Gorbunov, A.A. , H. Eichler, W. Pompe, B. Huey (1996) Lateral self-limitation in the laser-induced oxidation of ultrathin metal films. Appl Phys Lett 69: 2816–2818.
  26. Gorbunov, A.A., W. Pompe, H. Eichler, B. Huey, D.A. Bonell (1997) Nanostructuring of laser-deposited Ti films by self-limited oxidation. J Am Ceram Soc 80: 1663–1667.
  27. Gorbunov, A.A, A. Levin, D.C. Meyer, A. Tselev, P. Paufler, W. Pompe, E. Wieser (2002) Formation of unusual intermetallic phases by vacuum PLD. Appl Surf Sci 197–198: 392–397.
  28. Greenwood, J.A., J.E. Murphy-Ullrich (1998) Signaling of de-adhesion in cellular regulation and motility. Microsc Res Tech 43: 420–432.
  29. Grössner-Schreiber, B., M. Griepentrog, I. Haustein, W.D. Müller, K.P. Lange, H. Briedigkeit, U.B. Göbel (1991) Plaque formation on surface modified dental implants. An in vitro study. Clin Oral Implants Res 12: 543–551.
  30. Hatano, K., H. Inoue, T. Kojo, T. Matsunaga, T. Tsujisawa, Y. Uchiyama (1999) Effect of surface roughness on proliferation and alkaline phosphatase expression of rat calvarial cells cultured on polystyrene. Bone 25: 439–445.
  31. Hendrich, C., U. Noth, U. Stahl, F. Merklein, C.P. Rader, N. Schutze, R. Thull, R.S. Tuan, J. Eulert (2002) Testing of skeletal implant surfaces with human fetal osteoblasts. Clin Orthop 394: 278–289.
  32. Kieswetter, K., Z. Schwartz, D. D. Dean, B.D. Boyan (1996a) The role of implant surface characteristics in the healing of bone. Crit Rev Oral Biol Med 7: 329–345.
  33. Kieswetter, K., Z. Schwartz, T.W. Hummert, D.L. Cochran, J. Simpson, D.D. Dean, B.D. Boyan (1996b) Surface roughness modulates the local production of growth factors and cytokines by osteoblast-like MG-63 cells. J Biomed Mater Res 32: 55–63.
  34. Könönen, M., M. Hormia, J. Kwilahti, J. Hautaniemi, I Thesleff (1992) Effect of surface processing on the attachment, orientation and proliferation of human gingival fibroblasts on titanium. J Biomed Mater Res 26: 1325–1341.
  35. Long, M., H.J. Rack (1998) Titanium alloys in total joint replacement – a materials science perspective. Biomaterials 19: 1621–1639.
  36. Lu, X., Y. Leng (2003) Quantitative analysis of osteoblast behavior on microgrooved hydroxyapatite and titanium substrata. J Biomed Mater Res A 66: 677–687.

    External Resources

  37. Martin, J.Y., Z. Schwartz, T.W. Hummert, D.M. Schraub, J. Simpson, J. Lankford Jr., D.D. Dean, D.L. Cochran, D.D. Boyan (1995) Effect of titanium surface roughness on proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63). J Biomed Mater Res 29: 389–401.
  38. Martines, E., K. McGhee, C. Wilkinson, A. Curtis (2004) A parallel-plate flow chamber to study initial cell adhesion on a nanofeatured surface. IEEE Trans Nanobioscience 3: 90–95.

    External Resources

  39. McManus, A.J., R.H. Doremus, R.W. Siegel, R. Bizios (2005) Evaluation of cytocompatibility and bending modus of nanoceramic/polymer composites. J Biomed Mater Res A 72: 98–106.

    External Resources

  40. Mustafa, K., J. Wroblewski, K. Hultenby, B. Silva Lopez, K. Arvidson (2000) Effects of titanium surfaces blasted with TiO2 particles on the initial attachment of cells derived from human mandibular bone. A scanning electron microscopic and histomorphometric analysis. Clin Oral Implants Res 11: 116–128.
  41. Oakley, C., D. M. Brunette (1993) The sequence of alignment of microtubules, focal contacts and actin filament in fibroblasts spreading on smooth and grooved titanium substrata. J Cell Sci 106: 343–354.
  42. Ohgaki, M., T. Kizuki, M. Katsura, K. Yamashita (2001) Manipulation of selective cell adhesion and growth by surface charges of electrically polarized hydroxyapatite. J Biomed Mater Res 57: 366–373.
  43. Okumura, A., M. Goto, T. Goto, M. Yoshinari, S. Masuko, T. Katusuki, T. Tanaka (2001) Substrate affects the initial attachment and subsequent behavior of human osteoblastic cells (Saos-2). Biomaterials 22: 2263–2271.
  44. Ponta, H., L. Sherman, P.A. Herrlich (2003) CD44: from adhesion molecules to signaling regulators. Nat Rev Mol Cell Biol 4: 33–45.
  45. Price, R.L., K. Ellison, K.M. Haberstroh, T.J. Webster (2004) Nanometer surface roughness increases select osteoblast adhesion on carbon nanofiber compacts. J Biomed Mater Res A 70: 129–138.

    External Resources

  46. Puleo, D.A., L.A. Holleran, R.H. Doremus, R. Bizios (1991) Osteoblast responses to orthopedic implant materials in vitro. J Biomed Mater Res 25: 711–723.
  47. Rae, T. (1986) The biological response to titanium and titanium-aluminium-vanadium alloy particles. Biomaterials 7: 30–40.
  48. Robinson, K. R. (1985) The responses of cells to electric fields: a review. J Cell Biol 101: 2023–2027.
  49. Roehlecke, C., M. Witt, M., Kasper, E. Schulze, C. Wolf, A. Hofer, R.W. Funk (2001) Synergistic effect of titanium alloy and collagen type I on cell adhesion, proliferation and differentiation of osteoblast-like cells. Cells Tissues Organs 168: 178–187.
  50. Roessler, S., R. Zimmermann, D. Scharnweber, C. Werner, H. Worch (2002) Characterization of oxide layers on Ti6Al4V and titanium by streaming potential and streaming current measurements. Colloids Surf B Biointerfaces 26: 387–395.
  51. Rosa, A.L., M.M. Beloti (2003) Rat bone marrow cell response to titanium and titanium alloy with different surface roughness. Clin Oral Implants Res 14: 43–48.
  52. Ross, F.P., J. Chappel, J.I. Alvarez, D. Sander, W.T. Butler, M.C. Farach-Carson, K.A. Mintz, P.G. Robey, S.L. Teitelbaum, D.A. Cheresh (1993) Interaction between the bone matrix proteins osteopontin and bone sialoprotein and the osteoclast integrin alpha v beta 3 potentiate bone resorption. J Biol Chem 5: 9901–9907.

    External Resources

  53. Scutt, A., A. Berg, H. Mayer, H. (1992) A semiautomated 96-well plate assay for collagen synthesis. Anal Biochem 203: 209–294.

    External Resources

  54. Schwartz, Z., C.H. Lohmann, J. Oefinger, L.F. Bonewald, D.D. Dean, D.D. Boyan (1999) Implant surface characteristics modulate differential behavior of cells in the osteoblastic lineage. Adv Dent Res 13: 38:48.

    External Resources

  55. Soboyejo, W.O., B. Nemetski, S. Allameh, N. Marcantonio, C. Mercer, J. Ricci (2002) Interactions between MC3T3-E1 cells and textured Ti6Al4V surfaces. J Biomed Mater Res 62: 56–72.
  56. Teixeira, A.I., G.A. Abrahams, P.J. Bertics, C.J. Murphy, P.F. Nealey (2003) Epithelial contact guidance on well-defined micro- and nanostructured substrates. J Cell Sci 116: 1881–1892.
  57. Thompson, G.J., D.A. Puleo (1996) Ti-6Al-4V ion solution inhibition of osteogenic cell phenotype as a function of differentiation timecourse in vitro. Biomaterials 17: 1949–1954.
  58. Walboomers, X.F., H.J. Croes, L.A. Ginsel, J.A. Jansen (1998) Growth behavior of fibroblasts on microgrooved polysterene. Biomaterials 19: 1861–1868.
  59. Walboomers, X.F., J.A. Jansen (2001) Cell and tissue behavior on micro-grooved surfaces. Odontology 89: 2–11.
  60. Wang, E., M. Zaho, J.V. Forrester, C.D. McCaig, C.D. (2000a) Re-orientation and faster, directed migration of lens epithelial cells in a physiological electric field. Exp Eye Res 71: 91–98.
  61. Wang, J.H.C., E.S. Grood, J. Florer, R. Wenstrup (2000b) Alignment and proliferation of MC3T3-E1 osteoblasts in microgrooved silicone substrate subjected to cyclic stretching. J Biomech 33: 729–735.
  62. Weber, G.F., S. Ashkar, M.J. Glimcher, H. Cantor (1996) Receptor-ligand interaction between CD44 and osteopontin (Eta-1). Science 271: 509–512.
  63. Webster, T.J., C. Ergun, R.H. Doremus, R.W. Siegel, R. Bizios (2001) Enhanced osteoclast-like cell functions on nanophase ceramics. Biomaterials 22: 1327–1333.
  64. Webster, T.J., J.U. Ejiofor (2004) Increased osteoblast adhesion on nanophase metals: Ti, Ti6Al4V, and CoCrMo. Biomaterials 25: 4731–4739.
  65. Webster, T.J., E.L. Hellenmeyer, R.L. Price (2005) Increased osteoblast functions on theta + delta nanofiber alumina. Biomaterials 26: 953–960.
  66. Zohar, R., S. Cheifetz, C.A. McCulloch, J. Sodek (1998) Analysis of intracellular osteopontin as a marker of osteoblastic cell differentiation and mesenchymal cell migration. Eur J Oral Sci 106(suppl 1): 401–407.

    External Resources

  67. Zreiqat, H., T.N. Crotti, C.R. Howlett, M. Capone, B. Markovic, D.R. Haynes (2003) Prosthetic particles modify the expression of bone-related proteins by human osteoblastic cells in vitro. Biomaterials 24: 337–346.

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