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The impact of the RGD peptide on osteoblast adhesion and spreading on zinc-substituted hydroxyapatite surface

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Abstract

The incorporation of zinc into the hydroxyapatite structure (ZnHA) has been proposed to stimulate osteoblast proliferation and differentiation. Another approach to improve cell adhesion and hydroxyapatite (HA) performance is coating HA with adhesive proteins or peptides such as RGD (arginine–glycine–aspartic acid). The present study investigated the adhesion of murine osteoblastic cells to non-sintered zinc-substituted HA disks before and after the adsorption of RGD. The incorporation of zinc into the HA structure simultaneously changed the topography of disk’s surface on the nanoscale and the disk’s surface chemistry. Fluorescence microscopy analyses using RGD conjugated to a fluorescein derivative demonstrated that ZnHA adsorbed higher amounts of RGD than non-substituted HA. Zinc incorporation into HA promoted cell adhesion and spreading, but no differences in the cell density, adhesion and spreading were detected when RGD was adsorbed onto ZnHA. The pre-treatment of disks with fetal bovine serum (FBS) greatly increased the cell density and cell surface area for all RGD-free groups, overcoming the positive contribution of zinc to cell adhesion. The presence of RGD on the ZnHA surface impaired the effects of FBS pre-treatment possibly due to competition between FBS proteins and RGD for surface binding sites.

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Acknowledgments

The authors would like to thank CNPq, CAPES and FAPERJ for financial support. IME and LABNANO/CBPF for support with the SEM analysis; Jessica Dornelas, Suzana Anjos and Luciana Consentino for support with the in vitro tests and Priscila Lotsch for the mycoplasma verification assay. We also acknowledge the Brazilian Nanotechnology National Laboratory at Brazilian Center for Research in Energy and Materials for the electron microscopy facilities.

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Authors and Affiliations

  1. Brazilian Center for Physics Research, Xavier Sigaud 150, Urca, RJ, 22290-180, Brazil

    Elena Mavropoulos, Moema Hausen, Andrea M. Costa, Alexandre Mello & Alexandre M. Rossi

  2. Department of Cellular and Molecular Biology, Institute of Biology, UFF, Niterói, RJ, 24050-120, Brazil

    Gutemberg Alves

  3. Brazilian Nanotechnology National Laboratory (LNNano), Campinas, SP, 13083-970, Brazil

    C. A. Ospina

  4. National Institute of Metrology, N.S. das Graças 50, Duque de Caxias, RJ, CEP 25250-020, Brazil

    José M. Granjeiro

  5. Federal University of Alfenas, Exact Sciences Institute (ICEx) MG-Brasil, Alfenas, Brazil

    M. Mir

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  1. Elena Mavropoulos
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Correspondence to Elena Mavropoulos.

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Mavropoulos, E., Hausen, M., Costa, A.M. et al. The impact of the RGD peptide on osteoblast adhesion and spreading on zinc-substituted hydroxyapatite surface. J Mater Sci: Mater Med 24, 1271–1283 (2013). https://doi.org/10.1007/s10856-013-4851-3

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