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Fabrication of functional fibronectin patterns by nanosecond excimer laser direct write for tissue engineering applications

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Abstract

Laser direct write techniques represent a prospective alternative for engineering a new generation of hybrid biomaterials via the creation of patterns consisting of biological proteins onto practically any type of substrate. In this paper we report on the characterization of fibronectin features obtained onto titanium substrates by UV nanosecond laser transfer. Fourier-transform infrared spectroscopy measurements evidenced no modification in the secondary structure of the post-transferred protein. The molecular weight of the transferred protein was identical to the initial fibronectin, no fragment bands being found in the transferred protein’s Western blot migration profile. The presence of the cell-binding domain sequence and the mannose groups within the transferred molecules was revealed by anti-fibronectin monoclonal antibody immunolabelling and FITC-Concanavalin-A staining, respectively. The in vitro tests performed with MC3T3-E1 osteoblast-like cells and Swiss-3T3 fibroblasts showed that the cells’ morphology and spreading were strongly influenced by the presence of the fibronectin spots.

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Acknowledgments

The authors acknowledge with thanks the support of this work by Brancusi Programs n°14775SL (2006–2008) and n°25392UD (2011–2012). Romanian authors also acknowledge the partially funding support of the PCCA 153/02.07.2012 national project. We also thank Pr. Carmen Ristoscu and Pr. Emmanuel Pauthe for their valuable help, Pr. Patrick DiMartino for providing the lectins, Dr. Charlotte Vendrely and Dr. Damien Seyer for the useful explanations concerning the ATR-FTIR investigations, Dr. George Stan for the XRR Ti film thickness measurements and Dr. Rumeiza Bascetin for the useful discussions on fibronectin.

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Authors and Affiliations

  1. Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, Université de Cergy-Pontoise, 2 Av. Adolphe Chauvin, 95302, Cergy Pontoise, France

    S. Grigorescu, M. Hindié, F. Carreiras & O. Gallet

  2. National Institute for Lasers, Plasma and Radiation Physics, Str. Atomistilor Nr. 409, PO Box MG-36, 077125, Magurele, Ilfov, Romania

    S. Grigorescu, E. Axente & I. N. Mihailescu

  3. Institut de Science des Matériaux de Mulhouse, Université de Haute-Alsace, 15 rue Jean Starcky, BP 2488, 68057, Mulhouse, France

    K. Anselme

  4. Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, F2-027 Avenida Carlos Chagas Filho, 373, Ilha do Fundão, Rio de Janeiro, RJ, 21941-902, Brazil

    J. Werckmann

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  1. S. Grigorescu
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  2. M. Hindié
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Grigorescu, S., Hindié, M., Axente, E. et al. Fabrication of functional fibronectin patterns by nanosecond excimer laser direct write for tissue engineering applications. J Mater Sci: Mater Med 24, 1809–1821 (2013). https://doi.org/10.1007/s10856-013-4927-0

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