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The role of well-defined nanotopography of titanium implants on osseointegration: cellular and molecular events in vivo

Authors Karazisis D, Ballo A, Petronis S, Agheli H, Emanuelsson L, Thomsen P, Omar O

Received 25 November 2015

Accepted for publication 22 January 2016

Published 1 April 2016 Volume 2016:11 Pages 1367—1382

DOI https://doi.org/10.2147/IJN.S101294

Checked for plagiarism Yes

Review by Single-blind

Peer reviewers approved by Dr Govarthanan Muthusamy

Peer reviewer comments 3

Editor who approved publication: Dr Thomas J Webster


Dimitrios Karazisis,1–3 Ahmed M Ballo,1,2,4 Sarunas Petronis,2,5 Hossein Agheli,1,2 Lena Emanuelsson,1,2 Peter Thomsen,1,2 Omar Omar1,2

1Department of Biomaterials, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; 2BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden; 3Department of Oral and Maxillofacial Surgery, Sahlgrenska Academy, University of Gothenburg, Sweden; 4Department of Oral Health Sciences, Faculty of Dentistry, University of British Columbia, Vancouver, BC, Canada; 5Department of Chemistry, Materials and Surfaces, SP Technical Research Institute of Sweden, Borås, Sweden

Purpose: Mechanisms governing the cellular interactions with well-defined nanotopography are not well described in vivo. This is partly due to the difficulty in isolating a particular effect of nanotopography from other surface properties. This study employed colloidal lithography for nanofabrication on titanium implants in combination with an in vivo sampling procedure and different analytical techniques. The aim was to elucidate the effect of well-defined nanotopography on the molecular, cellular, and structural events of osseointegration.
Materials and methods: Titanium implants were nanopatterned (Nano) with semispherical protrusions using colloidal lithography. Implants, with and without nanotopography, were implanted in rat tibia and retrieved after 3, 6, and 28 days. Retrieved implants were evaluated using quantitative polymerase chain reaction, histology, immunohistochemistry, and energy dispersive X-ray spectroscopy (EDS).
Results: Surface characterization showed that the nanotopography was well defined in terms of shape (semispherical), size (79±6 nm), and distribution (31±2 particles/µm2). EDS showed similar levels of titanium, oxygen, and carbon for test and control implants, confirming similar chemistry. The molecular analysis of the retrieved implants revealed that the expression levels of the inflammatory cytokine, TNF-α, and the osteoclastic marker, CatK, were reduced in cells adherent to the Nano implants. This was consistent with the observation of less CD163-positive macrophages in the tissue surrounding the Nano implant. Furthermore, periostin immunostaining was frequently detected around the Nano implant, indicating higher osteogenic activity. This was supported by the EDS analysis of the retrieved implants showing higher content of calcium and phosphate on the Nano implants.
Conclusion: The results show that Nano implants elicit less periimplant macrophage infiltration and downregulate the early expression of inflammatory (TNF-α) and osteoclastic (CatK) genes. Immunostaining and elemental analyses show higher osteogenic activity at the Nano implant. It is concluded that an implant with the present range of well-defined nanocues attenuates the inflammatory response while enhancing mineralization during osseointegration.

Keywords: nanofabrication, gene expression, immunohistochemistry, energy dispersive X-ray spectroscopy, inflammatory cytokines, bone formation

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