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Open AccessArticle

Surface-Dependent Osteoblasts Response to TiO2 Nanotubes of Different Crystallinity

Ural Federal University, Mira Str. 19, 620002 Yekaterinburg, Russia
Institute of High-Temperature Electrochemistry of the Ural Branch of the Russian Academy of Sciences, Akademicheskaya Str. 20, 620990 Yekaterinburg, Russia
M.N. Mikheev Institute of Metal Physics of the Ural Branch of the Russian Academy of Sciences, Sofia Kovalevskaya Str. 18, 620219 Yekaterinburg, Russia
Leibniz Institute of Polymer Research Dresden, 01069 Dresden, Germany
Max Bergmann Center of Biomaterials Dresden, Hohe Str. 6, 01069 Dresden, Germany
Yekaterinburg Research Institute of Viral Infections, Rospotrebnadzor, Letnyaya Str. 23, 620030 Yekaterinburg, Russia
Institute of Chemical Technology and Engineering, Faculty of Chemical Technology, Poznan University of Technology, Berdychowo 4, 60965 Poznan, Poland
Authors to whom correspondence should be addressed.
Nanomaterials 2020, 10(2), 320;
Received: 16 December 2019 / Revised: 15 January 2020 / Accepted: 9 February 2020 / Published: 13 February 2020
(This article belongs to the Special Issue Nano-hybrids: Synthesis, Characterization and Applications)
One of the major challenges of implantology is to design nanoscale modifications of titanium implant surfaces inducing osseointegration. The aim of this study was to investigate the behavior of rat osteoblasts cultured on anodized TiO2 nanotubes of different crystallinity (amorphous and anatase phase) up to 24 days. TiO2 nanotubes were fabricated on VT1–0 titanium foil via a two-step anodization at 20 V using NH4F as an electrolyte. Anatase-phase samples were prepared by heat treatment at 500 °C for 1 h. VT1–0 samples with flat surfaces were used as controls. Primary rat osteoblasts were seeded over experimental surfaces for several incubation times. Scanning electron microscopy (SEM) was used to analyze tested surfaces and cell morphology. Cell adhesion and proliferation were investigated by cell counting. Osteogenic differentiation of cells was evaluated by qPCR of runt-related transcription factor 2 (RUNX2), osteopontin (OPN), integrin binding sialoprotein (IBSP), alkaline phosphatase (ALP) and osteocalcin (OCN). Cell adhesion and proliferation, cell morphology and the expression of osteogenic markers were affected by TiO2 nanotube layered substrates of amorphous and anatase crystallinity. In comparison with flat titanium, along with increased cell adhesion and cell growth a large portion of osteoblasts grown on the both nanostructured surfaces exhibited an osteocyte-like morphology as early as 48 h of culture. Moreover, the expression of all tested osteogenic markers in cells cultured on amorphous and anatase TiO2 nanotubes was upregulated at least at one of the analyzed time points. To summarize, we demonstrated that amorphous and anodized TiO2 layered substrates are highly biocompatible with rat osteoblasts and that the surface modification with about 1500 nm length nanotubes of 35 ± 4 (amorphous phase) and 41 ± 8 nm (anatase phase) in diameter is sufficient to induce their osteogenic differentiation. Such results are significant to the engineering of coating strategies for orthopedic implants aimed to establish a more efficient bone to implant contact and enhance bone repair.
Keywords: TiO2 nanotubes; anodization; amorphous; anatase; osseointegration; gene expression; implants TiO2 nanotubes; anodization; amorphous; anatase; osseointegration; gene expression; implants
MDPI and ACS Style

Khrunyk, Y.Y.; Belikov, S.V.; Tsurkan, M.V.; Vyalykh, I.V.; Markaryan, A.Y.; Karabanalov, M.S.; Popov, A.A.; Wysokowski, M. Surface-Dependent Osteoblasts Response to TiO2 Nanotubes of Different Crystallinity. Nanomaterials 2020, 10, 320.

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