Effect of nanoscale topography on fibronectin adsorption,focal adhesion size and matrix organisation |
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| Affiliation: | 1. Center for Biomaterials and Tissue Engineering, Universidad Politécnica de Valencia, 46022 Valencia, Spain;2. INEB – Instituto de Engenharia Biomédica, Div. Biomateriais, Porto, Portugal;3. ISEP – Instituto Superior de Engenharia do Porto, Dep. Engenharia Química, Porto, Portugal;4. Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Valencia, Spain;5. Regenerative Medicine Unit, Centro de Investigación Príncipe Felipe, Autopista del Saler 16, 46013 Valencia, Spain;1. Department of Biomedical Engineering, University of Minnesota – Twin Cities, Minneapolis, MN 55455, United States;2. Division of Cardiology, Department of Medicine, University of Minnesota – Twin Cities, Minneapolis, MN 55455, United States;1. Politehnica University Timişoara, Faculty of Industrial Chemistry and Environmental Engineering, 6 Pîrvan Blv., RO-300223, Timişoara, Romania;2. Politehnica University Timişoara, Research Institute for Renewable Energy, P-ta Victoriei No.2, 300006, Timişoara, Romania;3. “Vasile Goldis” Western University of Arad, The Institute of Life Sciences, No. 86, Liviu Rebreanu Street, 310414 Arad, Romania;1. Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA;2. Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA;3. Biophysics Graduate Group, Stanford University, Stanford, CA, 94305, USA;4. Department of Pediatrics, Stanford University, Stanford, CA, 94305, USA;5. Department of Materials Science and Engineering, Stanford University, Stanford, CA, 94305, USA;1. Department of Biomedical Engineering, National University of Singapore, Singapore;2. Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), Singapore;3. Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore;4. Duke-NUS Graduate Medical School, Singapore;5. Singapore National Eye Centre, Singapore;6. Department of Surgery, National University of Singapore, Singapore;7. Mechanobiology Institute, Singapore, National University of Singapore, Singapore;1. 3B''s Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal;2. ICVS/3B’s–PT Government Associate Laboratory, Braga/Guimarães, Portugal |
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| Abstract: | Phase separation of PLLA/PS (50/50, w/w) solutions during a spin-casting process gives rise to well-defined nanotopographies of 14, 29 and 45 nm deep pits depending on the concentration of the solution. Their influence on the biological activity of fibronectin (FN) was investigated. FN adsorption was quantified by radiolabelling the protein. The amount of adsorbed FN was higher on the 14 nm deep pit nanotopography than on the other two surfaces. FN distribution between valleys and peaks was investigated by AFM combined with image analysis. FN tends to adsorb preferentially on the valleys of the nanotopography only for the 14 nm system and when adsorbed from solutions of concentration lower than 10 μg/ml. Higher concentration of the FN solution leads to evenly distribution of the protein throughout the surface; moreover, there is no difference in the distribution of the protein between valleys and peaks for the other two systems (29 and 45 nm) irrespective of the concentration of the FN solution. The biological activity of the adsorbed protein layer was assessed by investigating MC3T3 osteoblast-like cells adhesion, FN reorganisation and late matrix formation on the different substrates. Even if initial cell adhesion is excellent for every substrate, the size of the focal adhesion plaques increases as the size of the pits in the nanotopography does. This is correlated to FN reorganisation, which only takes places on the 29 and 45 nm deep pits surfaces, where enhanced late matrix production was also found. |
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