Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium

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Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium Raheleh Miralami1, Laura Koepsell1, Thyagaseely Premaraj2, Bongok Kim3, Geoffrey M. Thiele4, J. Graham Sharp5, Kevin L. Garvin1, Fereydoon Namavar*1 1 Department of Orthopaedic Surgery and Rehabilitation, UNMC, Omaha, NE 2 Department of Growth & Development -, University of Nebraska-Lincoln, Lincoln, NE 3 Department of Adult Restorative Dentistry, University of Nebraska - Lincoln, Lincoln, NE 4 Department of Internal Medicine – Rheumatology, UNMC, Omaha, NE 5 Department of Genetics, Cell Biology & Anatomy, UNMC, Omaha, NE *Corresponding Author [email protected] ABSTRACT Titanium (Ti) is the material of choice for orthopaedic applications because it is biocompatible and encourages osteoblast ingrowth. It was shown that the biocompatibility of Ti metal is due to the presence of a thin native sub-stoichiometric titanium oxide layer which enhances the adsorption of mediating proteins on the surface [1]. The present studies were devised to evaluate the adhesion, survival, and growth of cells on the surface of new engineered nano-crystal films of titanium and titanium oxides and compare them with orthopaedic-grade titanium with microcrystals. The engineered nano-crystal films with hydrophilic properties are produced by employing an ion beam assisted deposition (IBAD) technique. IBAD combines physical vapor deposition with concurrent ion beam bombardment in a high vacuum environment to produce films (with 3 to 70 nm grain size) with superior properties. These films are “stitched” to the artificial orthopaedic implant materials with characteristics that affect the wettability and mechanical properties of the coatings. To characterize the biocompatibility of these nano-engineered surfaces, we have studied osteoblast function including cell adhesion, growth, and differentiation on different nanostructured samples. Cell responses to surfaces were examined using SAOS-2 osteoblast-like cells. We also studied a correlation between the surface nanostructures and the cell growth by characterizing the SAOS-2 cells with immunofluorescence and measuring the amount alizarin red concentration produced after 7 and 14 days. The number of adherent cells was determined by means of nuclei quantification on the nanocrystalline Ti, TiO2, and microcrystalline Ti and analysis was performed with Image J. Our experimental results indicated that nanocrystalline TiO2 is superior to both nano and microcrystalline Ti in supporting growth, adhesion, and proliferation. Improving the quality of surface oxide, i.e. fabricating stoichiometric oxides as well as nanoengineering the surface topology, is crucial for increasing the biocompatibility of Ti implant materials. INTRODUCTION Osseointegration of orthopaedic implants is dependent upon different parameters of the implant surface such as surface properties and physicochemical properties including surface energy, charge, wettability, chemistry, and topography. Surface topography can impact cellular beh