Nanopatterning was performed using colloidal lithography as described previously. Both types of implant were coated with a thin Ti film and heat-treated to provide identical surface chemistries. Implant surface modifications were made to produce two subsets of implants, a control implant with a machined surface (M implant) and a test implant with a topographically nanopatterned machined surface (MN implant). The dimensions were 2 mm in diameter and 5 mm in length. Screw-shaped implants made of grade II titanium (Ti) (Elos Medtech Pinol A/S, Denmark) were used. (30,31) Other additive techniques, such as colloidal lithography, (32,33) nanoimprinting, and replica molding, (22) provide possibilities to fabricate controlled nanopatterns even on complex three-dimensional (3D) implants. (28,29) In this context, the authors have emphasized the advantage of electrochemical anodization techniques for the fabrication of controlled nanopatterns while preserving microscale features of the underlying implant. Additive nanofabrication methods, on the other hand, are commonly based on low-voltage anodization process leading to partially ordered nanotube formation. (23,24) Lithographic techniques, using electron beam, (25) X-ray, (26) or laser (27) as the source of radiation, result in ordered nanopatterns, commonly applied on smooth and flat composite materials and wafers for in vitro studies. Acid etching is the most common subtractive method used for creating random nanotopographies on implants. The methods applied to produce nanopatterns can be categorized as either subtractive or additive. Strategies to enable the elucidation of such correlations on the nanoscale level include nanoprocessing techniques, allowing the experiments to be reproducible and consistent. (21) Generally, the resulting range of topographic features on multiple scales hinders the detailed understanding of the role of specific length scale surface properties in cellular, structural, biomechanical, and clinical outcomes. (20) The majority of these modifications entail subtractive techniques as blasting alone, blasting and etching or anodization in combinations. (17,19) Surface modifications have been an essential method by which material properties are optimized for oral implants. (15в€’18) The latter processes occur in a narrow interface zone between the surface of the material and tissue. This concept includes not only structural adaptation and/or bonding of organic and inorganic components but also the preceding cellular and molecular processes.

The science underpinning the long-term survival and success rates in the range of 80вЂ“99% for oral implants, (1в€’5) hip and knee arthroplasties, (6,7) amputation prostheses, (8в€’10) and bone-anchored hearing devices (11в€’14) goes back to the concept of osseointegration.

The implantation of materials in bone has revolutionized their application in orthopedic and cranio-maxillo-facial contexts. Controlled nanotopography, in the form of hemispherical 60 nm protrusions, promotes gene expressions related to early osteogenic differentiation and osteoblastic activity in implant-adherent cells in the human jaw bone. Correlation analysis reveals that older patient age is associated with increased expression of proinflammatory cytokines (TNF-О± and MCP-1) on the machined implants and decreased expression of pro-osteogenic factor (BMP-2) on the nanopatterned implants. No significant differences are observed in the expression of the analyzed inflammatory and remodeling genes. Compared to those adherent to the machined (M) implants, the cells adherent to the nanopatterned (MN) implants demonstrate significant upregulation (1.8- to 2-fold) of bone-related genes (RUNX2, ALP, and OC).

In the second-stage surgery, after 6вЂ“8 weeks, the mini-implants are retrieved by unscrewing, and the implant-adherent cells are subjected to gene expression analysis using quantitative polymerase chain reaction (qPCR).

When clinical implants are installed, additional two mini-implants, one with a machined surface (M) and one with a machined surface superimposed with a hemispherical nanopattern (MN), are installed in the posterior maxilla. Herein, the molecular response to an intentional, controlled nanotopography pattern superimposed on screw-shaped titanium implants is investigated in human bone. Implant surface modification by nanopatterning is an interesting route for enhancing osseointegration in humans.

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