Magnetic resonance imaging tracking of human adipose derived stromal cells within three-dimensional scaffolds for bone tissue engineering.

For bone tissue engineering, human Adipose Derived Stem Cells (hADSCs) are proposed to be associated with a scaffold for promoting bone regeneration. After implantation, cellularised scaffolds require a non-invasive method for monitoring their fate in vivo. The purpose of this study was to use Magnetic Resonance Imaging (MRI)-based tracking of these cells, labelled with magnetic agents for in vivo longitudinal assessment. hADSCs were isolated from adipose tissue and labelled with USPIO-rhodamine (Ultrasmall SuperParamagnetic Iron Oxide). USPIO internalisation, absence of toxicity towards hADSCs, and osteogenic differentiation of the labelled cells were evaluated in standard culture conditions. Labelled cells were then seeded within a 3D porous polysaccharide-based scaffold and imaged in vitro using fluorescence microscopy and MRI. Cellularised scaffolds were implanted subcutaneously in nude mice and MRI analyses were performed from 1 to 28 d after implantation. In vitro, no effect of USPIO labelling on cell viability and osteogenic differentiation was found. USPIO were efficiently internalised by hADSCs and generated a high T2* contrast. In vivo MRI revealed that hADSCs remain detectable until 28 d after implantation and could migrate from the scaffold and colonise the area around it. These data suggested that this scaffold might behave as a cell carrier capable of both holding a cell fraction and delivering cells to the site of implantation. In addition, the present findings evidenced that MRI is a reliable technique to validate cell-seeding procedures in 3D porous scaffolds, and to assess the fate of hADSCs transplanted in vivo.

[1]  Jean-Christophe Ginefri,et al.  High-resolution 1.5-Tesla magnetic resonance imaging for tissue-engineered constructs: a noninvasive tool to assess three-dimensional scaffold architecture and cell seeding. , 2010, Tissue engineering. Part C, Methods.

[2]  Jeff W M Bulte,et al.  Clinically applicable labeling of mammalian and stem cells by combining superparamagnetic iron oxides and transfection agents. , 2003, Radiology.

[3]  S. Primack,et al.  High-resolution CT: normal anatomy, techniques, and pitfalls. , 2001, Radiologic clinics of North America.

[4]  G. Hossein-Zadeh,et al.  Quantitative evaluation of optimal imaging parameters for single-cell detection in MRI using simulation. , 2010, Magnetic resonance imaging.

[5]  Nicolas Grenier,et al.  In vivo MR imaging of intravascularly injected magnetically labeled mesenchymal stem cells in rat kidney and liver. , 2004, Radiology.

[6]  K. Shakesheff,et al.  Engineering embryonic stem-cell aggregation allows an enhanced osteogenic differentiation in vitro. , 2010, Tissue engineering. Part C, Methods.

[7]  Etienne Duguet,et al.  A method for synthesis and functionalization of ultrasmall superparamagnetic covalent carriers based on maghemite and dextran , 2005 .

[8]  G. Vunjak‐Novakovic,et al.  Tissue engineered bone grafts: biological requirements, tissue culture and clinical relevance. , 2008, Current stem cell research & therapy.

[9]  Miya Ishihara,et al.  Osteogenic Potential of Human Adipose Tissue-Derived Stromal Cells as an Alternative Stem Cell Source , 2004, Cells Tissues Organs.

[10]  Heather Kalish,et al.  Efficient magnetic cell labeling with protamine sulfate complexed to ferumoxides for cellular MRI. , 2004, Blood.

[11]  J. Rubin,et al.  The Osteogenic Potential of Adipose-Derived Stem Cells for the Repair of Rabbit Calvarial Defects , 2006, Annals of plastic surgery.

[12]  B. Tomanek,et al.  Adipose-derived stem cells are an effective cell candidate for treatment of heart failure: an MR imaging study of rat hearts. , 2009, American journal of physiology. Heart and circulatory physiology.

[13]  F. Chaubet,et al.  Fabrication of porous polysaccharide-based scaffolds using a combined freeze-drying/cross-linking process. , 2010, Acta biomaterialia.

[14]  L. Politi MR-based imaging of neural stem cells , 2007, Neuroradiology.

[15]  H. de Boer The history of bone grafts. , 1988, Clinical orthopaedics and related research.

[16]  Fabien Hyafil,et al.  Ferumoxtran-10–Enhanced MRI of the Hypercholesterolemic Rabbit Aorta: Relationship Between Signal Loss and Macrophage Infiltration , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[17]  A. Boccaccini,et al.  Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. , 2006, Biomaterials.

[18]  P. Wielopolski,et al.  Effects of iron oxide incorporation for long term cell tracking on MSC differentiation in vitro and in vivo. , 2008, Biochemical and biophysical research communications.

[19]  Bindiya Patel,et al.  Adipose-derived stem cells: isolation, expansion and differentiation. , 2008, Methods.

[20]  D. Kaplan,et al.  Porosity of 3D biomaterial scaffolds and osteogenesis. , 2005, Biomaterials.

[21]  N. Selvamurugan,et al.  Biocomposites containing natural polymers and hydroxyapatite for bone tissue engineering. , 2010, International journal of biological macromolecules.

[22]  Jean-Marie Devoisselle,et al.  Magnetic nanoparticles and their applications in medicine. , 2006, Nanomedicine.

[23]  K. Kusumoto,et al.  Bone tissue engineering using human adipose-derived stem cells and honeycomb collagen scaffold. , 2008, Journal of biomedical materials research. Part A.

[24]  J. Franconi,et al.  4D retrospective black blood trueFISP imaging of mouse heart , 2009, Magnetic resonance in medicine.

[25]  Ackerman Ga Substituted naphthol AS phosphate derivatives for the localization of leukocyte alkaline phosphatase activity. , 1962 .

[26]  R. Bizios,et al.  Engineering bone: challenges and obstacles , 2005, Journal of cellular and molecular medicine.

[27]  Matthew D. Kwan,et al.  Cell-based therapies for skeletal regenerative medicine. , 2008, Human molecular genetics.

[28]  H. D. Boer THE HISTORY OF BONE GRAFTS , 1988 .

[29]  Isabelle Raynal,et al.  Macrophage Endocytosis of Superparamagnetic Iron Oxide Nanoparticles: Mechanisms and Comparison of Ferumoxides and Ferumoxtran-10 , 2004, Investigative radiology.

[30]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[31]  Byung-Soo Kim,et al.  In vivo bone formation following transplantation of human adipose-derived stromal cells that are not differentiated osteogenically. , 2008, Tissue engineering. Part A.

[32]  G. Vunjak‐Novakovic,et al.  Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. , 2010, Tissue engineering. Part A.

[33]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[34]  A. Arbab,et al.  Labeling of cells with ferumoxides–protamine sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells , 2005, NMR in biomedicine.

[35]  H. D. de Vries,et al.  Comparison of SPIO and USPIO for in vitro labeling of human monocytes: MR detection and cell function. , 2007, Radiology.

[36]  C. Stehling,et al.  Comparative study of imaging at 3.0 T versus 1.5 T of the knee , 2009, Skeletal Radiology.

[37]  Jeff W M Bulte,et al.  In Vivo Magnetic Resonance Tracking of Magnetically Labeled Cells after Transplantation , 2002, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[38]  L. Pénicaud,et al.  Adipose tissues display differential phagocytic and microbicidal activities depending on their localization , 2001, International Journal of Obesity.

[39]  J. Gimble,et al.  Adipose-derived stem cells for regenerative medicine. , 2007, Circulation research.

[40]  Craig H Meyer,et al.  Technology Insight: in vivo cell tracking by use of MRI , 2006, Nature Clinical Practice Cardiovascular Medicine.

[41]  C. Dani,et al.  A role for preadipocytes as macrophage‐like cells , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[42]  Jeff W M Bulte,et al.  Feridex labeling of mesenchymal stem cells inhibits chondrogenesis but not adipogenesis or osteogenesis , 2004, NMR in biomedicine.

[43]  E. Gontier,et al.  Study of the MR relaxation of microglia cells labeled with Gd-DTPA-bearing nanoparticles. , 2009, Contrast media & molecular imaging.

[44]  Reine Bareille,et al.  Human endothelial progenitor cell attachment to polysaccharide-based hydrogels: A pre-requisite for vascular tissue engineering , 2007, Journal of materials science. Materials in medicine.

[45]  K. Brown,et al.  Bone and cartilage transplantation in orthopaedic surgery. A review. , 1982, The Journal of bone and joint surgery. American volume.

[46]  Wee Keong Nah,et al.  The osteogenic differentiation of adipose tissue-derived precursor cells in a 3D scaffold/matrix environment. , 2008, Current drug discovery technologies.

[47]  J. Franconi,et al.  Nanoparticle phagocytosis and cellular stress: involvement in cellular imaging and in gene therapy against glioma , 2010, NMR in biomedicine.

[48]  D G Nishimura,et al.  Linear combination steady‐state free precession MRI , 2000, Magnetic resonance in medicine.

[49]  Chris Heyn,et al.  In vivo magnetic resonance imaging of single cells in mouse brain with optical validation , 2006, Magnetic resonance in medicine.

[50]  E. Hsu,et al.  Superparamagnetic iron oxide labeling and transplantation of adipose-derived stem cells in middle cerebral artery occlusion-injured mice. , 2007, AJR. American journal of roentgenology.

[51]  J. Deux,et al.  Iron oxide nanoparticle-labeled rat smooth muscle cells: cardiac MR imaging for cell graft monitoring and quantitation. , 2005, Radiology.

[52]  D. Letourneur,et al.  Pullulan-based hydrogel for smooth muscle cell culture. , 2007, Journal of biomedical materials research. Part A.

[53]  M. Hedrick,et al.  Fat tissue: an underappreciated source of stem cells for biotechnology. , 2006, Trends in biotechnology.

[54]  U Himmelreich,et al.  Efficient stem cell labeling for MRI studies. , 2008, Contrast media & molecular imaging.

[55]  Ardeshir Bayat,et al.  Adult stem cells in tissue engineering , 2009, Expert review of medical devices.