Mesenchymal stem cells from patients to assay bone graft substitutes

Authors

  • M. Manfrini,

    1. Section of Cell Biology and Molecular Genetics, Department of Morphology, Surgery and Experimental Medicine, School of Medicine and Surgery, University of Ferrara, Ferrara, Italy
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  • C. Di Bona,

    1. Section of Cell Biology and Molecular Genetics, Department of Morphology, Surgery and Experimental Medicine, School of Medicine and Surgery, University of Ferrara, Ferrara, Italy
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  • A. Canella,

    1. Department of Biochemistry and Molecular Biology, University of Ferrara, Ferrara, Italy
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  • E. Lucarelli,

    1. Laboratory of Bone Regeneration, Istituti Ortopedici Rizzoli, Bologna, Italy
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  • A. Pellati,

    1. Section of Histology and Embryology, Department of Morphology, Surgery and Experimental Medicine, School of Medicine and Surgery, University of Ferrara, Ferrara, Italy
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  • A. D'Agostino,

    1. Department of Odontostomatology and Maxillo-Facial Surgery, School of Medicine and School of Dentistry, University of Verona, Verona, Italy
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  • G. Barbanti-Bròdano,

    1. Spine Surgery, Istituto Ortopedico “Rizzoli,” Bologna, Italy
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  • M. Tognon

    Corresponding author
    1. Section of Cell Biology and Molecular Genetics, Department of Morphology, Surgery and Experimental Medicine, School of Medicine and Surgery, University of Ferrara, Ferrara, Italy
    • Section of Cell Biology and Molecular Genetics, Department of Morphology, Surgery and Experimental Medicne, School of Medicine and Surgery, University of Ferrara, V. Fossato di Mortara 64/b, Ferrara 44121, Italy.
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  • Authors declare no conflict of interest.

Abstract

Bio-engineered scaffolds used in orthopedic clinical applications induce different tissue responses after implantation. In this study, non-stoichiometric Mg2+ ions and stoichiometric apatites, which are used in orthopedic surgery as bone substitutes, have been assayed in vitro with human adult mesenchymal stem cells (hMSC) to evaluate cytocompatibility and osteoconductivity. hMSCs from the bone marrow aspirates of orthopedic patients were isolated and analyzed by flow cytometry for the surface markers Stro1, CD29, CD44, CD71, CD73, CD90, CD105 (positive) and CD45, CD235 (negative). The hMSC were analyzed for self-renewal capacity and for differentiation potential. The hMSC, which were grown on different biomaterials, were analyzed for (i) cytotoxicity by AlamarBlue metabolic assay, (ii) osteoconductivity by ELISA for activated focal adhesion kinase, (iii) cytoskeleton organization by fluorescence microscopy, and (iv) cell morphology which was investigated by scan electron microscopy (SEM). Results indicate that isolated cell populations agree with minimal criteria for defining hMSC cultures. Non-stoichiometric Mg2+ and stoichiometric apatites, in granular form, represent a more favorable environment for mesenchymal stem cell adhesion and growth compared to the non-stoichiometric Mg2+ apatite, in nano-structured paste form. This study indicates that different forms of biomaterials modulate osteoconductivity and cellular growth by differential activation focal adhesion kinase. J. Cell. Physiol. 228: 1229–1237, 2013. © 2012 Wiley Periodicals, Inc.

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