OBJECTIVE: Cell therapies have emerged as a promising approach in medicine. The basis of each therapy is the injection of 1-100×10(6) cells with regenerative potential into some part of the body. Mesenchymal stromal cells (MSCs) are the most used cell type in the cell therapy nowadays, but no gold standard for the labeling of the MSCs for magnetic resonance imaging (MRI) is available yet. This work evaluates our newly synthesized uncoated superparamagnetic maghemite nanoparticles (surface-active maghemite nanoparticles - SAMNs) as an MRI contrast intracellular probe usable in a clinical 1.5 T MRI system. METHODS: MSCs from rat and human donors were isolated, and then incubated at different concentrations (10-200 μg/mL) of SAMN maghemite nanoparticles for 48 hours. Viability, proliferation, and nanoparticle uptake efficiency were tested (using fluorescence microscopy, xCELLigence analysis, atomic absorption spectroscopy, and advanced microscopy techniques). Migration capacity, cluster of differentiation markers, effect of nanoparticles on long-term viability, contrast properties in MRI, and cocultivation of labeled cells with myocytes were also studied. RESULTS: SAMNs do not affect MSC viability if the concentration does not exceed 100 μg ferumoxide/mL, and this concentration does not alter their cell phenotype and long-term proliferation profile. After 48 hours of incubation, MSCs labeled with SAMNs show more than double the amount of iron per cell compared to Resovist-labeled cells, which correlates well with the better contrast properties of the SAMN cell sample in T2-weighted MRI. SAMN-labeled MSCs display strong adherence and excellent elasticity in a beating myocyte culture for a minimum of 7 days. CONCLUSION: Detailed in vitro tests and phantom tests on ex vivo tissue show that the new SAMNs are efficient MRI contrast agent probes with exclusive intracellular uptake and high biological safety.

Department of Comparative Biomedicine and Food Science University of Padua Padova Italy

Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

Department of Medical Chemistry and Biochemistry Faculty of Medicine Palacky University Olomouc Czech Republic

Department of Pharmacology Masaryk University Brno Czech Republic

Regional Centre of Advanced Technologies and Materials Department of Physical Chemistry and Analytical Chemistry Faculty of Science Palacky University Olomouc Czech Republic

Regional Centre of Advanced Technologies and Materials Department of Physical Chemistry and Analytical Chemistry Faculty of Science Palacky University Olomouc Czech Republic ; Department of Comparative Biomedicine and Food Science University of Padua Padova Italy

Zobrazit více v PubMed

Martin-Rendon E, Brunskill SJ, Hyde CJ, Stanworth SJ, Mathur A, Watt SM. Autologous bone marrow stem cells to treat acute myocardial infarction: a systematic review. Eur Heart J. 2008;29:1807–1818. PubMed

Wakitani S, Mitsuoka T, Nakamura N, Toritsuka Y, Nakamura Y, Horibe S. Autologous bone marrow stromal cell transplantation for repair of full-thickness articular cartilage defects in human patellae: two case reports. Cell Transplant. 2004;13:595–600. PubMed

Kebriaei P, Robinson S. Treatment of graft-versus-host-disease with mesenchymal stromal cells. Cytotherapy. 2011;13:262–268. PubMed

Wang Z, Ruan J, Cui DX. Advances and prospect of nanotechnology in stem cells. Nanoscale Res Lett. 2009;4:593–605. PubMed PMC

Corot C, Robert P, Idée JM, Port M. Recent advances in iron oxide nanocrystal technology for medical imaging. Adv Drug Deliv Rev. 2006;58:1471–1504. PubMed

Kraitchman DL, Heldman AW, Atalar E, et al. In vivo magnetic resonance imaging of mesenchymal stem cells in myocardial infarction. Circulation. 2003;107:2290–2293. PubMed

Berman SMC, Walczak P, Bulte JWM. Tracking stem cells using magnetic nanoparticles. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2011;3:343–355. PubMed PMC

Bulte JWM, Kraitchman DL. Iron oxide MR contrast agents for molecular and cellular imaging. NMR Biomed. 2004;17:484–499. PubMed

Pankhurst QA, Connolly J, Jones SK, et al. Applications of magnetic nanoparticles in biomedicine. J Phys D Appl Phys. 2003;36:R167–R181.

Wang YXJ, Hussain SM, Krestin GP. Superparamagnetic iron oxide contrast agents: physicochemical characteristics and applications in MR imaging. Eur Radiol. 2001;11:2319–2331. PubMed

Weissleder R, Moore A, Mahmood U, et al. In vivo magnetic resonance imaging of transgene expression. Nat Med. 2000;6:351–355. PubMed

Berry CC, Curtis ASG. Functionalisation of magnetic nanoparticles for applications in biomedicine. J Phys D Appl Phys. 2003;36:198–206.

Alexiou C, Jurgons R, Seliger G, Iro H. Medical applications of magnetic nanoparticles. J Nanosci Nanotechnol. 2006;6:2762–2768. PubMed

Laurent S, Forge D, Port M, et al. Magnetic iron oxide nanoparticles: synthesis, stabilization, vectorization, physicochemical characterization, and biological applications. Chem Rev. 2008;108:2064–2110. PubMed

Thanh NTK. Magnetic Nanoparticles from Fabrication to Clinical Applications. Boca Raton: CRC Press, Taylor & Francis; 2012.

Sinigaglia G, Magro M, Miotto G, et al. Catalytically active bovine serum amine oxidase bound to fluorescent and magnetically drivable nanoparticles. Int J Nanomedicine. 2012;7:2249–2259. PubMed PMC

Venerando R, Miotto G, Magro M, et al. Magnetic nanoparticles with covalently bound self-assembled protein corona for advanced biomedical applications. J Phys Chem C. 2013;117:20320–20331.

Magro M, Sinigaglia G, Nodari L, et al. Charge binding of rhodamine derivative to OH− stabilized nanomaghemite: universal nanocarrier for construction of magnetofluorescent biosensors. Acta Biomater. 2012;8:2068–2076. PubMed

Gallo MP, Ramella R, Alloatti G, et al. Limited plasticity of mesenchymal stem cells cocultured with adult cardiomyocytes. J Cell Biochem. 2007;100:86–99. PubMed

Chlopcikova S, Psotova J, Miketova P. Neonatal rat cardiomyocytes – a model for the study of morphological, biochemical and electrophysiological characteristics of the heart. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2001;145:49–55. PubMed

Tucek J, Zboril R, Petridis D. Maghemite nanoparticles by view of Mossbauer spectroscopy. J Nanosci Nanotechnol. 2006;6:926–947. PubMed

Shapiro EM, Skrtic S, Sharer K, Hill JM, Dunbar CE, Koretsky AP. MRI detection of single particles for cellular imaging. Proc Natl Acad Sci U S A. 2004;101:10901–10906. PubMed PMC

Smirnov P, Gazeau F, Beloeil JC, Doan BT, Wilhelm C, Gillet B. Single-cell detection by gradient echo 9.4 T MRI: a parametric study. Contrast Media Mol Imaging. 2006;1:165–174. PubMed

Kostura L, Kraitchman DL, Mackay AM, Pittenger MF, Bulte JW. Feridex labeling of mesenchymal stem cells inhibits chondrogenesis but not adipogenesis or osteogenesis. NMR Biomed. 2004;17:513–517. PubMed

Chang YK, Liu YP, Ho JH, Hsu SC, Lee OK. Aminesurface-modified superparamagnetic iron oxide nanoparticles interfere with differentiation of human mesenchymal stem cells. J Orthop Res. 2003;30:1499–1506. PubMed

Magro M, Faralli A, Baratella D, et al. Avidin functionalized maghemite nanoparticles and their application for recombinant human biotinyl-SERCA purification. Langmuir. 2012;28:15392–15401. PubMed

da Rocha EL, Caramoni GF, Rambo CR. Nanoparticle translocation through a lipid bilayer tuned by surface chemistry. Phys Chem Chem Phys. 2013;15:2282–2290. PubMed

Stark WJ. Nanoparticles in biological systems. Angew Chem Int Ed Engl. 2011;50:1242–1258. PubMed

Singh N, Jenkins G, Asadi R. Potential toxicity of supermagnetic iron oxide. Nano Rev. 2010;1:538–553. PubMed PMC

Haruhs-Frenkel O, Debotton N, Benita S, Altschuler Y. Targeting of nanoparticles to the clathrin-mediated endocytic pathway. Biochem Biophys Res Commun. 2007;353:26–32. PubMed

Kralj S, Rojnik M. Effect of surface charge on the cellular uptake of fluorescent magnetic nanoparticles. J Nanopart Res. 2012;14(10):1–14. PubMed

Jo JI, Aoki I, Tabata Y. Design of iron oxide nanoparticles with different sizes and surface charges for simple and efficient labeling of mesenchymal stem cells. J Control Release. 2010;142(3):465–473. PubMed

Bhattarai SR, Kim SY, Jang K. Laboratory formulated magnetic nanoparticles for enhancement of viral gene expression in suspension cell line. J Virol Methods. 2008;147(2):213–218. PubMed

Pradhan P, Giri J, Banerjee R. Cellular interactions of lauric acid and dextran-coated magnetite nanoparticles. J Magn Magn Mater. 2007;311(1):282–287.

Luciani N, Gazeau F, Wilhelm C. Reactivity of the monocyte/macrophage system to superparamagnetic anionic nanoparticles. J Mater Chem. 2009;19(35):6373–6380.

Wilhelm C, Gazeau F, Roger J, et al. Interaction of anionic superparamagnetic nanoparticles with cells: kinetic analysis of membrane adsorption and subsequent internalization. Langmuir. 2002;18:8148–8155.

Golovko DM, Henning T, Bauer JS, et al. Accelerated stem cell labeling with ferucarbotran and protamine. Eur Radiol. 2010;20:640–648. PubMed PMC

The Effect of Rhodamine-Derived Superparamagnetic Maghemite Nanoparticles on the Motility of Human Mesenchymal Stem Cells and Mouse Embryonic Fibroblast Cells

The surface reactivity of iron oxide nanoparticles as a potential hazard for aquatic environments: A study on Daphnia magna adults and embryos

Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells