Ti-6Al-4V-based nanotubes were prepared on a Ti-6Al-4V surface by anodic oxidation on 10 V, 20 V, and 30 V samples. The 10 V, 20 V, and 30 V samples and a control smooth Ti-6Al-4V sample were evaluated in terms of their chemical composition, diameter distribution, and cellular response. The surfaces of the 10 V, 20 V, and 30 V samples consisted of nanotubes of a relatively wide range of diameters that increased with the voltage. Saos-2 cells had a similar initial adhesion on all nanotube samples to the control Ti-6Al-4V sample, but it was lower than on glass. On day 3, the highest concentrations of both vinculin and talin measured by enzyme-linked immunosorbent assay and intensity of immunofluorescence staining were on 30 V nanotubes. On the other hand, the highest concentrations of ALP, type I collagen, and osteopontin were found on 10 V and 20 V samples. The final cellular densities on 10 V, 20 V, and 30 V samples were higher than on glass. Therefore, the controlled anodization of Ti-6Al-4V seems to be a useful tool for preparing nanostructured materials with desirable biological properties.

Department of Biomaterials and Tissue Engineering Institute of Physiology Czech Academy of Sciences Prague Czech Republic

Department of Metals and Corrosion Engineering University of Chemistry and Technology Prague Czech Republic

Zobrazit více v PubMed

Candel JJ, Amigo V. Recent advances in laser surface treatment of titanium alloys. J Laser Appl. 2011;23(2):022005.

Guo J, Padilla RJ, Ambrose W, De Kok IJ, Cooper LF. The effect of hydrofluoric acid treatment of TiO2 grit blasted titanium implants on adherent osteoblast gene expression in vitro and in vivo. Biomaterials. 2007;28(36):5418–5425. PubMed

Chu CL, Hu T, Zhou J, et al. Effects of H2O2 pretreatment on surface characteristics and bioactivity of NaOH-treated NiTi shape memory alloy. Trans Nonferrous Metal Soc China. 2006;16(6):1295–1300.

Kim HW, Koh YH, Li LH, Lee S, Kim HE. Hydroxyapatite coating on titanium substrate with titania buffer layer processed by sol-gel method. Biomaterials. 2004;25(13):2533–2538. PubMed

Lu X, Zhao Z, Leng Y. Biomimetic calcium phosphate coatings on nitric-acid-treated titanium surfaces. Mater Sci Eng C Mater Biol Appl. 2007;27(4):700–708.

Tamilselvi S, Raghavendran HB, Srinivasan P, Rajendran N. In vitro and in vivo studies of alkali- and heat-treated Ti-6Al-7Nb and Ti-5Al-2Nb-1Ta alloys for orthopedic implants. J Biomed Mater Res A. 2009;90(2):380–386. PubMed

Nebe B, Finke B, Lüthen F, et al. Improved initial osteoblast functions on amino-functionalized titanium surfaces. Biomol Eng. 2007;24(5):447–454. PubMed

Balakrishnan M, Narayanan R. Synthesis of anodic titania nanotubes in Na2SO4/NaF electrolyte: a comparison between anodization time and specimens with biomaterial based approaches. Thin Solid Films. 2013;540:23–30.

Gregory CA, Gunn WG, Peister A, Prockop DJ. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem. 2004;329(1):77–84. PubMed

Li Y, Ding D, Ning C, et al. Thermal stability and in vitro bioactivity of Ti-Al-V-O nanostructures fabricated on Ti6Al4V alloy. Nanotechnology. 2009;20(6):065708. PubMed

Macak JM, Tsuchiya H, Taveira L, Ghicov A, Schmuki P. Self-organized nanotubular oxide layers on Ti-6A1-7Nb and Ti-6A1-4V formed by anodization in NH4F solutions. J Biomed Mater Res A. 2005;75A(4):928–933. PubMed

Oh S, Brammer KS, Li YS, et al. Stem cell fate dictated solely by altered nanotube dimension. Proc Natl Acad Sci U S A. 2009;106(7):2130–2135. PubMed PMC

Sjöström T, Lalev G, Mansell JP, Su B. Initial attachment and spreading of MG63 cells on nanopatterned titanium surfaces via through-mask anodization. Appl Surf Sci. 2011;257(10):4552–4558.

Brammer KS, Oh S, Cobb CJ, Bjursten LM, van der Heyde H, Jin S. Improved bone-forming functionality on diameter-controlled TiO2 nanotube surface. Acta Biomater. 2009;5(8):3215–3223. PubMed

Brammer KS, Oh S, Frandsen CJ, Varghese S, Jin S. Nanotube surface triggers increased chondrocyte extracellular matrix production. Mater Sci Eng C Mater Biol Appl. 2010;30(4):518–525.

Park J, Bauer S, Schlegel KA, Neukam FW, von der Mark K, Schmuki P. TiO2 nanotube surfaces: 15 nm – an optimal length scale of surface topography for cell adhesion and differentiation. Small. 2009;5(6):666–671. PubMed

Matejka R. ALICE: fluorescent image analyser (version 1.0) [software] [Accessed August 24, 2015]. Available from: http://alice.fbmi.cvut.cz.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265–275. PubMed

National Institute of Standards and Technology NIST standard reference database online database list: Free standard reference databases by SRD number. 2015. [Accessed August 7, 2015]. Available from: http://www.nist.gov/srd/onlinelist.cfm.

Habazaki H, Fushimi K, Shimizu K, Skeldon P, Thompson GE. Fast migration of fluoride ions in growing anodic titanium oxide. Electrochem Commun. 2007;9(5):1222–1227.

Wang L, Zhao TT, Zhang Z, Li G. Fabrication of highly ordered TiO2 nanotube arrays via anodization of Ti-6Al-4V alloy sheet. J Nanosci Nanotechnol. 2010;10(12):8312–8321. PubMed

Moon SH, Lee SJ, Park IS, et al. Bioactivity of Ti-6Al-4V alloy implants treated with ibandronate after the formation of the nanotube TiO2 layer. J Biomed Mater Res B Appl Biomater. 2012;100(8):2053–2059. PubMed

Pérez-Jorge C, Conde A, Arenas MA, et al. In vitro assessment of Staphylococcus epidermidis and Staphylococcus aureus adhesion on TiO2 nanotubes on Ti-6Al-4V alloy. J Biomed Mater Res A. 2012;100(7):1696–1705. PubMed

Neacsu P, Mazare A, Cimpean A, et al. Reduced inflammatory activity of RAW 264.7 macrophages on titania nanotube modified Ti surface. Int J Biochem Cell Biol. 2014;55:187–195. PubMed

Sista S, Nouri A, Li Y, Wen C, Hodgson PD, Pande G. Cell biological responses of osteoblasts on anodized nanotubular surface of a titanium-zirconium alloy. J Biomed Mater Res A. 2013;101(12):3416–3430. PubMed

Ross AP, Webster TJ. Anodizing color coded anodized Ti6Al4V medical devices for increasing bone cell functions. Int J Nanomedicine. 2013;8:109–117. PubMed PMC

Park JW, Kim HK, Kim YJ, Jang JH, Song H, Hanawa T. Osteoblast response and osseointegration of a Ti-6Al-4V alloy implant incorporating strontium. Acta Biomater. 2010;6(7):2843–2851. PubMed

Nguyen TD, Moon SH, Oh TJ, Park IS, Lee MH, Bae TS. The effect of APH treatment on surface bonding and osseointegration of Ti-6Al-7Nb implants: an in vitro and in vivo study. J Biomed Mater Res B Appl Biomater. 2014;103(3):641–648. PubMed

Weiner S, Wagner HD. The material bone: structure-mechanical function relations. Annu Rev Mater Sci. 1998;28:271–298.

Kane R, Ma PX. Mimicking the nanostructure of bone matrix to regenerate bone. Mater Today (Kidlington) 2013;16(11):418–423. PubMed PMC

Minagar S, Li Y, Berndt CC, Wen C. The influence of titania-zirconia-zirconium titanate nanotube characteristics on osteoblasts cell adhesion. Acta Biomater. 2015;12:281–289. PubMed

Lan MY, Liu CP, Huang HH, Lee SW. Both enhanced biocompatibility and antimicrobial activity in Ag-decorated TiO2 nanotubes. PLoS One. 2013;8(10):e75364. PubMed PMC

Czekanska EM, Stoddart MJ, Richards RG, Hayes JS. In search of an osteoblast cell model for in vitro research. Eur Cell Mater. 2012;24:1–17. PubMed

Pautke C, Schieker M, Tischer T, et al. Characterization of osteosarcoma cell lines MG-63, Saos-2 cells and U-2 OS in comparison to human osteoblasts. Anticancer Res. 2004;24(6):3743–3748. PubMed

Rodan SB, Imai Y, Thiede MA, et al. Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Res. 1987;47(18):4961–4966. PubMed

Vandrovcova M, Jirka I, Novotna K, et al. Interaction of human osteoblast-like cells and MG-63 cells with thermally oxidized surfaces of a titanium-niobium alloy. Plos One. 2014;9(6):e100475. PubMed PMC

Ramaglia L, Postiglione L, Di Spigna G, Capece G, Salzano S, Rossi G. Sandblasted-acid-etched titanium surface influences in vitro the biological behavior of SaOS-2 human osteoblast-like cells. Dent Mater J. 2011;30(2):183–192. PubMed

Minagar S, Li Y, Berndt CC, Wen C. The influence of titania-zirconia-zirconium titanate nanotube characteristics on osteoblast cell adhesion. Acta Biomater. 2015;12:281–289. PubMed

Hempel U, Hefti T, Kalbacova M, Wolf-Brandstetter C, Dieter P, Schlottig F. Response of osteoblast-like SAOS-2 cells to zirconia ceramics with different surface topographies. Clin Oral Implants Res. 2010;21(2):174–181. PubMed

Solař P, Kylián O, Marek A, et al. Particles induced surface nanoroughness of titanium surface and its influence on adhesion of osteoblast-like MG-63 cells. Appl Surf Sci. 2015;324:99–105.

Vandrovcova M, Hanus J, Drabik M, et al. Effect of different surface nanoroughness of titanium dioxide films on the growth of human osteoblast-like MG63 cells. J Biomed Mater Res A. 2012;100(4):1016–1032. PubMed

Park J, Bauer S, von der Mark K, Schmuki P. Nanosize and vitality: TiO2 nanotube diameter directs cell fate. Nano Lett. 2007;7(6):1686–1691. PubMed

Yu WQ, Jiang XQ, Zhang FQ, Xu L. The effect of anatase TiO2 nanotube layers on MC3T3-E1 preosteoblast adhesion, proliferation, and differentiation. J Biomed Mater Res A. 2010;94(4):1012–1022. PubMed

Kulkarni M, Mazare A, Gongadze E, et al. Titanium nanostructures for biomedical applications. Nanotechnology. 2015;26(6):062002. PubMed

Bauer S, Park J, Faltenbacher J, Berger S, von der Mark K, Schmuki P. Size selective behavior of mesenchymal stem cells on ZrO2 and TiO2 nanotube arrays. Integr Biol (Camb) 2009;1(8–9):525–532. PubMed

Anselme K, Ploux L, Ponche A. Cell/material interfaces: influence of surface chemistry and surface topography on cell adhesion. J Adhes Sci Technol. 2010;24(5):831–852.

Gongadze E, Kabaso D, Bauer S, et al. Adhesion of osteoblasts to a nanorough titanium implant surface. Int J Nanomedicine. 2011;6:1801–1816. PubMed PMC

Webster TJ, Ergun C, Doremus RH, Siegel RW, Bizios R. Specific proteins mediate enhanced osteoblast adhesion on nanophase ceramics. J Biomed Mater Res. 2000;51(3):475–483. PubMed

Gongadze E, Kabaso D, Bauer S, Park J, Schmuki P, Iglič A. Adhesion of osteoblasts to a vertically aligned TiO2 nanotube surface. Mini Rev Med Chem. 2013;13(2):194–200. PubMed

Peng Z, Ni J, Zheng K, et al. Dual effects and mechanism of TiO2 nanotube arrays in reducing bacterial colonization and enhancing C3H10T1/2 cell adhesion. Int J Nanomedicine. 2013;8:3093–3105. PubMed PMC

Wang X, Yan C, Ye K, He Y, Li Z, Ding J. Effect of RGD nanospacing on differentiation of stem cells. Biomaterials. 2013;34(12):2865–2874. PubMed

Lv L, Liu Y, Zhang P, et al. The nanoscale geometry of TiO2 nanotubes influences the osteogenic differentiation of human adipose-derived stem cells by modulating H3K4 trimethylation. Biomaterials. 2015;39:193–205. PubMed

Hu Y, Cai K, Luo Z, et al. TiO2 nanotubes as drug nanoreservoirs for the regulation of mobility and differentiation of mesenchymal stem cells. Acta Biomater. 2012;8(1):439–448. PubMed

Son WW, Zhu X, Shin HI, Ong JL, Kim KH. In vivo histological response to anodized and anodized/hydrothermally treated titanium implants. J Biomed Mater Res B Appl Biomater. 2003;66(2):520–525. PubMed

Enhancement of biocompatibility of anodic nanotube structures on biomedical Ti-6Al-4V alloy via ultrathin TiO2 coatings

A Ru/RuO2-Doped TiO2 Nanotubes as pH Sensors for Biomedical Applications: The Effect of the Amount and Oxidation of Deposited Ru on the Electrochemical Response

Mesenchymal stem cell interaction with Ti6Al4V alloy pre-exposed to simulated body fluid

Different diameters of titanium dioxide nanotubes modulate Saos-2 osteoblast-like cell adhesion and osteogenic differentiation and nanomechanical properties of the surface

Adhesion and differentiation of Saos-2 osteoblast-like cells on chromium-doped diamond-like carbon coatings

Silver Nanocoating Technology in the Prevention of Prosthetic Joint Infection