title: Mesenchymal stem cells from patients to assay bone graft substitutes.
abstract: Bio-engineered scaffolds used in orthopedic clinical applications induce different tissue responses after implantation. In this study, non-stoichiometric Mg(2+) 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 Mg(2+) and stoichiometric apatites, in granular form, represent a more favorable environment for mesenchymal stem cell adhesion and growth compared to the non-stoichiometric Mg(2+) 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.
authors.affiliation: Section of Cell Biology and Molecular Genetics, Department of Morphology, Surgery and Experimental Medicine, School of Medicine and Surgery, University of Ferrara, Ferrara, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Manfrini
authors.forename: C
authors.initials: C
authors.lastname: Di Bona
authors.forename: A
authors.initials: A
authors.lastname: Canella
authors.forename: E
authors.initials: E
authors.lastname: Lucarelli
authors.forename: A
authors.initials: A
authors.lastname: Pellati
authors.forename: A
authors.initials: A
authors.lastname: D'Agostino
authors.forename: G
authors.initials: G
authors.lastname: Barbanti-Bròdano
authors.forename: M
authors.initials: M
authors.lastname: Tognon
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism* / Q000637:transplantation / Q000648:ultrastructure; D001672:Biocompatible Materials; D015415:Biomarkers / Q000378:metabolism; D001861:Bone Regeneration; D018786:Bone Substitutes; D016025:Bone Transplantation / Q000379:methods; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D017886:Durapatite / Q000378:metabolism; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D006801:Humans; D008274:Magnesium / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000378:metabolism* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D053758:Nanoparticles; D010766:Phosphorylation; D011208:Powders; D013997:Time Factors; D054457:Tissue Scaffolds; D014443:Tyrosine
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D018786:Bone Substitutes; D011208:Powders; D014443:Tyrosine; D017886:Durapatite; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D008274:Magnesium