Skip to main content

Advertisement

Log in

In vitro assays for adhesion and migration of osteoblastic cells (Saos-2) on titanium surfaces

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

The first event occurring at the boundary between a metal implant and living tissue is the attachment of cells onto the metal surface of the implant. The attachment characteristics of the metal in this situation are critical in determining its biocompatibility and usefulness as artificial bone and tooth implants. Using the human osteosarcoma cell line Saos-2, we attempted to establish simple and reliable methods for evaluating the attachment of cultured osteoblastic cells onto titanium samples that had been subjected to various surface treatments. Fluorescence actin imaging showed that cells cultured on titanium with hydrofluoric acid etching (HF-Ti) exhibited delayed spreading of their cytoplasm, as compared to cells cultured for the same length of time on nitrided titanium or physically polished titanium. The HF-Ti-cultured cells also exhibited poor assembly of focal contacts, as visualized by vinculin immunofluorescence. Furthermore, in motility assays based on an in vitro wound model, cells cultured on HF-Ti migrated more slowly than cells cultured on other titanium surfaces. These data suggest that Saos-2 cells attach less effectively to the HF-Ti surface. The methods described in this study should be useful for assessing the initial interactions of cultured cells with various materials, including metals.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ahmad M, Gawronski D, Blum J, Goldberg J, Gronowicz G (1999) Differential response of human osteoblast-like cells to commercially pure (cp) titanium grades 1 and 4. J Biomed Mater Res 46:121–131

    Article  PubMed  CAS  Google Scholar 

  • Anselme K, Linez P, Bigerelle M, Le Maguer D, Le Maguer A, Hardouin P, Hildebrand HF, Iost A, Leroy JM (2000) The relative influence of the topography and chemistry of TiAl6V4 surfaces on osteoblastic cell behaviour. Biomaterials 21:1567–1577

    Article  PubMed  CAS  Google Scholar 

  • Baxter LC, Frauchiger V, Textor M, Gwynn I ap, Richards RG (2002) Fibroblast and osteoblast adhesion and morphology on calcium phosphate surfaces. Eur Cell Mater 30:1–17

    Google Scholar 

  • Brighton CT, Albelda SM (1992) Identification of integrin cell-substratum adhesion receptors on cultured rat bone cells. J Orthop Res 10:766–773

    Article  PubMed  CAS  Google Scholar 

  • Brunette DM, Chehroudi B (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

    Article  PubMed  CAS  Google Scholar 

  • Degasne I, Basle MF, Demais V, Hure G, Lesourd M, Grolleau B, Mercier L, Chappard D (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

    Article  PubMed  CAS  Google Scholar 

  • Deligianni DD, Katsala N, Ladas S, Sotiropoulou D, Amedee J, Missirlis YF (2001) Effect of surface roughness of the titanium alloy Ti-6Al-4V on human bone marrow cell response and on protein adsorption. Biomaterials 22:1241–1251

    Article  PubMed  CAS  Google Scholar 

  • Gao SY, Li CY, Chen J, Pan L, Saito S, Terashita T, Saito K, Miyawaki K, Shigemoto K, Mominoki K, Matsuda S, Kobayashi N (2004) Rho-ROCK signal pathway regulates microtubule-based process formation of cultured podocytes-inhibition of ROCK promoted process elongation. Nephron Exp Nephrol 97:e49–e61

    Article  PubMed  CAS  Google Scholar 

  • Geckeler K, Wacker R, Martini F, Hack A, Aicher W (2003) Enhanced biocompatibility for SAOS-2 osteosarcoma cells by surface coating with hydrophobic epoxy resins. Cell Physiol Biochem 13:155–164

    Article  PubMed  CAS  Google Scholar 

  • Goto T, Yoshinari M, Kobayashi S, Tanaka T (2004) The initial attachment and subsequent behavior of osteoblastic cells and oral epithelial cells on titanium. Biomed Mater Eng 14:537–544

    PubMed  Google Scholar 

  • Groessner-Schreiber B, Neubert A, Muller WD, Hopp M, Griepentrog M, Lange KP (2003) Fibroblast growth on surface-modified dental implants: an in vitro study. J Biomed Mater Res [A] 64:591–599

    Article  Google Scholar 

  • Gutmanas EY, Gotman I (2004) PIRAC Ti nitride coated Ti-6AI-4V head against UHMWPE acetabular cup-hip wear simulator study. J Mater Sci Mater Med 15:327–330

    Article  PubMed  CAS  Google Scholar 

  • Huang HH, Ho CT, Lee TH, Lee TL, Liao KK, Chen FL (2004) Effect of surface roughness of ground titanium on initial cell adhesion. Biomol Eng 21:93–97

    Article  PubMed  CAS  Google Scholar 

  • Kawahara H, Soeda Y, Niwa K, Takahashi M, Kawahara D, Araki N (2004) In vitro study on bone formation and surface topography from the standpoint of biomechanics. J Mater Sci Mater Med 15:1297–1307

    Article  PubMed  CAS  Google Scholar 

  • Keller JC (1998) Tissue compatibility to different surfaces of dental implants: in vitro studies. Implant Dent 7:331–337

    Article  PubMed  CAS  Google Scholar 

  • Knabe C, Klar F, Fitzner R, Radlanski RJ, Gross U (2002) In vitro investigation of titanium and hydroxyapatite dental implant surfaces using a rat bone marrow stromal cell culture system. Biomaterials 23:3235–3245

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi N, Mominoki K, Wakisaka H, Shimazaki Y, Matsuda S (2001) Morphogenetic activity of extracellular matrices on cultured podocytes. Laminin accelerates podocyte process formation in vitro. Ital J Anat Embryol 106 (2 Suppl 1):423–430

    PubMed  CAS  Google Scholar 

  • Lange R, Luthen F, Beck U, Rychly J, Baumann A, Nebe B (2002) Cell-extracellular matrix interaction and physico-chemical characteristics of titanium surfaces depend on the roughness of the material. Biomol Eng 19:255–261

    Article  PubMed  CAS  Google Scholar 

  • Leven RM, Virdi AS, Sumner DR (2004) Patterns of gene expression in rat bone marrow stromal cells cultured on titanium alloy discs of different roughness. J Biomed Mater Res [A] 70:391–401

    Article  Google Scholar 

  • Nobes CD, Hall A (1999) Rho GTPases control polarity, protrusion, and adhesion during cell movement. J Cell Biol 144:1235–1244

    Article  PubMed  CAS  Google Scholar 

  • Okumura A, Goto M, Goto T, Yoshinari M, Masuko S, Katsuki T, Tanaka T (2001) Substrate affects the initial attachment and subsequent behavior of human osteoblastic cells (Saos-2). Biomaterials 22:2263–2271

    Article  PubMed  CAS  Google Scholar 

  • Owen GR, Meredith DO, Gwynn I ap, Richard RG (2005) Focal adhesion quantification—a new assay of material biocompatibility? Eur Cell Mater 9:85–96

    PubMed  CAS  Google Scholar 

  • Scarano A, Piattelli M, Vrespa G, Caputi S, Piattelli A (2003) Bacterial adhesion on titanium nitride-coated and uncoated implants: an in vivo human study. J Oral Implantol 29:80–85

    Article  PubMed  Google Scholar 

  • ter Brugge PJ, Dieudonne S, Jansen JA (2002) Initial interaction of U2OS cells with noncoated and calcium phosphate coated titanium substrates. J Biomed Mater Res 61:399–407

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The English in this paper was checked by two professional readers, both native speakers of English.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Naoto Kobayashi.

Additional information

S.G. is the recipient of a grant awarded to foreign students by the government of Japan. This study was supported by the Integrated Center for Science (INCS) at Ehime University.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, Cy., Gao, Sy., Terashita, T. et al. In vitro assays for adhesion and migration of osteoblastic cells (Saos-2) on titanium surfaces. Cell Tissue Res 324, 369–375 (2006). https://doi.org/10.1007/s00441-005-0153-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-005-0153-5

Keywords

Navigation