Due to the bio-inert nature of titanium (Ti) and subsequent accompanying chronic inflammatory response, an implant's stability and function can be significantly affected, which is why various surface modifications have been employed, including the deposition of titanium oxide (TiO2) nanotubes (TNTs) onto the native surface through the anodic oxidation method. While the influence of nanotube diameter on cell behaviour and osteogenesis is very well documented, information regarding the effects of nanotube lateral spacing on the in vivo new bone formation process is insufficient and hard to find. Considering this, the present study's aim was to evaluate the mechanical properties and the osteogenic ability of two types of TNTs-based pins with different lateral spacing, e.g., 25 nm (TNTs) and 92 nm (spTNTs). The mechanical properties of the TNT-coated implants were characterised from a morphological point of view (tube diameter, spacing, and tube length) using scanning electron microscopy (SEM). In addition, the chemical composition of the implants was evaluated using X-ray photoelectron spectroscopy, while surface roughness and topography were characterised using atomic force microscopy (AFM). Finally, the implants' hardness and elastic modulus were investigated using nanoindentation measurements. The in vivo new bone formation was histologically evaluated (haematoxylin and eosin-HE staining) at 6 and 30 days post-implantation in a rat model. Mechanical characterisation revealed that the two morphologies presented a similar chemical composition and mechanical strength, but, in terms of surface roughness, the spTNTs exhibited a higher average roughness. The microscopic examination at 1 month post-implantation revealed that spTNTs pins (57.21 ± 34.93) were capable of promoting early new bone tissue formation to a greater extent than the TNTs-coated implants (24.37 ± 6.5), with a difference in the average thickness of the newly formed bone tissue of ~32.84 μm, thus highlighting the importance of this parameter when designing future dental/orthopaedic implants.

Department of Biochemistry and Molecular Biology Faculty of Biology University of Bucharest 91 95 Spl Independentei 050657 Bucharest Romania

Department of Materials Science and Engineering Chair of General Materials Properties Friedrich Alexander University of Erlangen Nürnberg Martensstraße 5 91058 Erlangen Germany

Department of Materials Science and Engineering WW4 LKO Friedrich Alexander University of Erlangen Nürnberg Martensstrasse 7 91058 Erlangen Germany

Faculty of Veterinary Medicine University of Agronomic Sciences and Veterinary Medicine 105 Spl Independentei 050097 Bucharest Romania

Regional Centre of Advanced Technologies and Materials Šlechtitelů 27 78371 Olomouc Czech Republic

Research Institute of the University of Bucharest University of Bucharest 050657 Bucharest Romania

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