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      Nanometer Scale Titanium Surface Texturing Are Detected by Signaling Pathways Involving Transient FAK and Src Activations

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          Abstract

          Background

          It is known that physico/chemical alterations on biomaterial surfaces have the capability to modulate cellular behavior, affecting early tissue repair. Such surface modifications are aimed to improve early healing response and, clinically, offer the possibility to shorten the time from implant placement to functional loading. Since FAK and Src are intracellular proteins able to predict the quality of osteoblast adhesion, this study evaluated the osteoblast behavior in response to nanometer scale titanium surface texturing by monitoring FAK and Src phosphorylations.

          Methodology

          Four engineered titanium surfaces were used for the study: machined (M), dual acid-etched (DAA), resorbable media microblasted and acid-etched (MBAA), and acid-etch microblasted (AAMB). Surfaces were characterized by scanning electron microscopy, interferometry, atomic force microscopy, x-ray photoelectron spectroscopy and energy dispersive X-ray spectroscopy. Thereafter, those 4 samples were used to evaluate their cytotoxicity and interference on FAK and Src phosphorylations. Both Src and FAK were investigated by using specific antibody against specific phosphorylation sites.

          Principal Findings

          The results showed that both FAK and Src activations were differently modulated as a function of titanium surfaces physico/chemical configuration and protein adsorption.

          Conclusions

          It can be suggested that signaling pathways involving both FAK and Src could provide biomarkers to predict osteoblast adhesion onto different surfaces.

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          Most cited references 36

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          Effects of titanium surface topography on bone integration: a systematic review.

          To analyse possible effects of titanium surface topography on bone integration. Our analyses were centred on a PubMed search that identified 1184 publications of assumed relevance; of those, 1064 had to be disregarded because they did not accurately present in vivo data on bone response to surface topography. The remaining 120 papers were read and analysed, after removal of an additional 20 papers that mainly dealt with CaP-coated and Zr implants; 100 papers remained and formed the basis for this paper. The bone response to differently configurated surfaces was mainly evaluated by histomorphometry (bone-to-implant contact), removal torque and pushout/pullout tests. A huge number of the experimental investigations have demonstrated that the bone response was influenced by the implant surface topography; smooth (S(a) 1-2 microm) surfaces showed stronger bone responses than rough (S(a)>2 microm) in some studies. One limitation was that it was difficult to compare many studies because of the varying quality of surface evaluations; a surface termed 'rough' in one study was not uncommonly referred to as 'smooth' in another; many investigators falsely assumed that surface preparation per se identified the roughness of the implant; and many other studies used only qualitative techniques such as SEM. Furthermore, filtering techniques differed or only height parameters (S(a), R(a)) were reported. * Surface topography influences bone response at the micrometre level. * Some indications exist that surface topography influences bone response at the nanometre level. * The majority of published papers present an inadequate surface characterization. * Measurement and evaluation techniques need to be standardized. * Not only height descriptive parameters but also spatial and hybrid ones should be used.
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            Mediation of biomaterial-cell interactions by adsorbed proteins: a review.

            An appropriate cellular response to implanted surfaces is essential for tissue regeneration and integration. It is well described that implanted materials are immediately coated with proteins from blood and interstitial fluids, and it is through this adsorbed layer that cells sense foreign surfaces. Hence, it is the adsorbed proteins, rather than the surface itself, to which cells initially respond. Diverse studies using a range of materials have demonstrated the pivotal role of extracellular adhesion proteins--fibronectin and vitronectin in particular--in cell adhesion, morphology, and migration. These events underlie the subsequent responses required for tissue repair, with the nature of cell surface interactions contributing to survival, growth, and differentiation. The pattern in which adhesion proteins and other bioactive molecules adsorb thus elicits cellular reactions specific to the underlying physicochemical properties of the material. Accordingly, in vitro studies generally demonstrate favorable cell responses to charged, hydrophilic surfaces, corresponding to superior adsorption and bioactivity of adhesion proteins. This review illustrates the mediation of cell responses to biomaterials by adsorbed proteins, in the context of osteoblasts and selected materials used in orthopedic implants and bone tissue engineering. It is recognized, however, that the periimplant environment in vivo will differ substantially from the cell-biomaterial interface in vitro. Hence, one of the key issues yet to be resolved is that of the interface composition actually encountered by osteoblasts within the sequence of inflammation and bone regeneration.
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              On implant surfaces: a review of current knowledge and opinions.

              The aims of the present review are (1) to identify essential surface parameters; (2) to present an overview of surface characteristics at the micrometer and nanometer levels of resolution relevant for the four most popular oral implant systems; (3) to discuss potential advantages of nanoroughness, hydrophilicity, and biochemical bonding; and (4) to suggest a hypothetical common mechanism behind strong bone responses to novel implant surfaces from different commercial companies. Oral implants from four major companies varied in average surface roughness (Sa) from 0.3 to 1.78 microm and in the developed surface area ratio (Sdr) from 24% to 143%, with the smoothest implants originating from Biomet 3i and the roughest from Institut Straumann. The original Branemark turned, machined surface had an Sa of 0.9 microm and an Sdr of 34%, making it clearly rougher than the smoothest implants examined. When evaluated for nanometer roughness, there was a substantial variation in Sa in the different implants from the four major companies. Novel implants from Biomet 3i, AstraTech, and Straumann differed from their respective predecessors in microroughness, physicochemical properties, and nano_roughness. When examined with scanning electron microscopy at high magnification, it was noted that these novel implant surfaces all had particular nanoroughness structures that were not present in their respective predecessors; this finding was suggested as a possible common mechanism behind the demonstrated stronger bone responses to these implants compared to adequate controls.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                7 July 2014
                : 9
                : 7
                Affiliations
                [1 ]Departmento de Química e Bioquímica, Instituto de Biociências, Universidade Estadual Paulista - UNESP, Botucatu, São Paulo, Brazil
                [2 ]Faculdade de Odontologia de Bauru, Universidade de São Paulo, Bauru, São Paulo, Brazil
                [3 ]Department of Prosthodontics, Faculty of Odontology, Malmö University, Malmö, Sweden
                [4 ]Department of Chemical and Biological Engineering, Applied Surface Chemistry, Chalmers University of Technology, Gothenburg, Sweden
                [5 ]Department of Cell and Molecular Biology, Institute of Biology, Universidade Federal Fluminense, Niteroi, Brazil
                [6 ]Excellion Biomedical Services, Petrópolis, Rio de Janeiro, Brazil
                [7 ]National Institute of Metrology, Quality and Technology - INMETRO, Xerém, Rio de Janeiro, Brazil
                [8 ]Department of Biomaterials and Biomimetics/Director for Research Department of Periodontology and Implant Dentistry, New York University College of Dentistry, New York, New York, United States of America
                Université de Technologie de Compiègne, France
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: WFZ EAB JMG ERT. Performed the experiments: WFZ EAB RJ MH MA GA PJB. Analyzed the data: WFZ EAB GA PJB RJ MH MA ERT. Contributed reagents/materials/analysis tools: WFZ PGC JMG. Wrote the paper: WFZ ERT PGC GA.

                Article
                PONE-D-13-29671
                10.1371/journal.pone.0095662
                4085036
                24999733

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Pages: 11
                Funding
                This study was funded by Fapesp, CNPq and Faperj. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Physical Sciences
                Chemistry
                Chemical Properties
                Physicochemical Properties
                Physical Chemistry
                Materials Science
                Material Properties
                Materials Characterization
                Materials Design
                Metallurgy
                Biology and Life Sciences
                Biotechnology
                Biomaterials
                Cell Biology
                Cell Processes
                Cell Growth
                Cell Adhesion
                Molecular Cell Biology
                Genetics
                Gene Expression

                Uncategorized

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