Magneto-mechanical Stimulation of Bone Marrow Mesenchymal Stromal Cells for Chondrogenic Differentiation Studies

Document Type : Research Paper


1 Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran

2 National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

3 Advanced Micro and Nano Devices Lab, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran


Mechanical interaction of cells and their surroundings are prominent in mechanically active tissues such as cartilage. Chondrocytes regulate their growth, matrix synthesis, metabolism, and differentiation in response to mechanical loadings. Cells sense and respond to applied physical forces through mechanosensors such as integrin receptors. Herein, we examine the role of mechanical stimulation of integrins in regards to their mechanotransduction ability to promote chondrogenesis. For this purpose, magnetic nanoparticles were chemically bonded to cell membrane mechanoreceptors and stimulated. Histological results showed the endocytosis of nanoparticles over the experimental period, pointing out the inefficient mechanical stimulation of the mechanoreceptors. Moreover, gene expression analysis only showed significant upregulation for SOX9, whereas type II collagen and aggrecan gene expression were not significantly different from the control group. Our results suggest that magneto-mechanical stimulation studies using magnetic nanoparticles should not only focus on the mechanical aspects, but also the interaction of magnetic nanoparticles with intracellular machinery should be investigated as well.


Main Subjects

[1]          A. J. Sophia Fox, A. Bedi, and S. A. Rodeo, “The basic science of articular cartilage: structure, composition, and function,” Sports health, vol. 1, no. 6, pp. 461–468, 2009.
[2]          D. Chen, J. Shen, W. Zhao, T. Wang, L. Han, J. L. Hamilton, and H.-J. Im, “Osteoarthritis: toward a comprehensive understanding of pathological mechanism,” Bone research, vol. 5, p. 16044, 2017.
[3]          P. D. Tatman, W. Gerull, S. Sweeney-Easter, J. I. Davis, A. O. Gee, and D.-H. Kim, “Multiscale biofabrication of articular cartilage: bioinspired and biomimetic approaches,” Tissue Engineering Part B: Reviews, vol. 21, no. 6, pp. 543–559, 2015.
[4]          E. B. Hunziker, “Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects,” Osteoarthritis and cartilage, vol. 10, no. 6, pp. 432–463, 2002.
[5]          H. K. Heywood, G. Nalesso, D. A. Lee, and F. Dell’Accio, “Culture expansion in low-glucose conditions preserves chondrocyte differentiation and enhances their subsequent capacity to form cartilage tissue in three-dimensional culture,” BioResearch open access, vol. 3, no. 1, pp. 9–18, 2014.
[6]          A. I. Caplan, “Mesenchymal stem cells,” Journal of orthopaedic research, vol. 9, no. 5, pp. 641–650, 1991.
[7]          H. Koga, L. Engebretsen, J. E. Brinchmann, T. Muneta, and I. Sekiya, “Mesenchymal stem cell-based therapy for cartilage repair: a review,” Knee Surgery, Sports Traumatology, Arthroscopy, vol. 17, no. 11, pp. 1289–1297, 2009.
[8]          A. O. Oseni, C. Crowley, M. Z. Boland, P. E. Butler, and A. M. Seifalian, “Cartilage tissue engineering: the application of nanomaterials and stem cell technology,” in Tissue engineering for tissue and organ regeneration, InTech, 2011.
[9]          N. Fahy, M. Alini, and M. J. Stoddart, “Mechanical stimulation of mesenchymal stem cells: Implications for cartilage tissue engineering,” Journal of Orthopaedic Research®, vol. 36, no. 1, pp. 52–63, 2018.
[10]        L. A. McMahon, F. J. O’Brien, and P. J. Prendergast, “Biomechanics and mechanobiology in osteochondral tissues,” 2008.
[11]        C. Vinatier, D. Mrugala, C. Jorgensen, J. Guicheux, and D. Noël, “Cartilage engineering: a crucial combination of cells, biomaterials and biofactors,” Trends in biotechnology, vol. 27, no. 5, pp. 307–314, 2009.
[12]        G. Musumeci, “The effect of mechanical loading on articular cartilage.” Multidisciplinary Digital Publishing Institute, 2016.
[13]        J. Sanchez-Adams, H. A. Leddy, A. L. McNulty, C. J. O’Conor, and F. Guilak, “The mechanobiology of articular cartilage: bearing the burden of osteoarthritis,” Current rheumatology reports, vol. 16, no. 10, pp. 451, 2014.
[14]        S. J. Millward-Sadler and D. M. Salter, “Integrin-dependent signal cascades in chondrocyte mechanotransduction,” Annals of biomedical engineering, vol. 32, no. 3, pp. 435–446, 2004.
[15]        A. N. Gasparski and K. A. Beningo, “Mechanoreception at the cell membrane: more than the integrins,” Archives of biochemistry and biophysics, vol. 586, pp. 20–26, 2015.
[16]        R. F. Loeser, “Integrins and chondrocyte–matrix interactions in articular cartilage,” Matrix Biology, vol. 39, pp. 11–16, 2014.
[17]        P. Roca-Cusachs, T. Iskratsch, and M. P. Sheetz, “Finding the weakest link–exploring integrin-mediated mechanical molecular pathways,” J Cell Sci, vol. 125, no. 13, pp. 3025–3038, 2012.
[18]        J. Cores, T. G. Caranasos, and K. Cheng, “Magnetically Targeted Stem Cell Delivery for Regenerative Medicine,” Journal of functional biomaterials, vol. 6, no. 3, pp. 526–546, 2015.
[19]        Y. Gao, J. Lim, S.-H. Teoh, and C. Xu, “Emerging translational research on magnetic nanoparticles for regenerative medicine,” Chemical Society reviews, vol. 44, no. 17, pp. 6306–6329, 2015.
[20]        H.-C. Kim, E. Kim, S. W. Jeong, T.-L. Ha, S.-I. Park, S. G. Lee, S. J. Lee, and S. W. Lee, “Magnetic nanoparticle-conjugated polymeric micelles for combined hyperthermia and chemotherapy,” Nanoscale, vol. 7, no. 39, pp. 16470–16480, 2015.
[21]        M. H. Cho, E. J. Lee, M. Son, J.-H. Lee, D. Yoo, J. Kim, S. W. Park, J.-S. Shin, and J. Cheon, “A magnetic switch for the control of cell death signalling in in vitro and in vivo systems,” Nature materials, vol. 11, no. 12, p. 1038, 2012.
[22]        J. W. M. Bulte, T. Douglas, B. Witwer, S.-C. Zhang, E. Strable, B. K. Lewis, H. Zywicke, B. Miller, P. van Gelderen, and B. M. Moskowitz, “Magnetodendrimers allow endosomal magnetic labeling and in vivo tracking of stem cells,” Nature biotechnology, vol. 19, no. 12, p. 1141, 2001.
[23]        J. M. Kanczler, H. S. Sura, J. Magnay, D. Green, R. O. C. Oreffo, J. P. Dobson, and A. J. El Haj, “Controlled differentiation of human bone marrow stromal cells using magnetic nanoparticle technology,” Tissue Engineering Part A, vol. 16, no. 10, pp. 3241–3250, 2010.
[24]        C. Monzel, C. Vicario, J. Piehler, M. Coppey, and M. Dahan, “Magnetic control of cellular processes using biofunctional nanoparticles,” Chemical science, vol. 8, no. 11, pp. 7330–7338, 2017.
[25]        J. R. Henstock, M. Rotherham, H. Rashidi, K. M. Shakesheff, and A. J. El Haj, “Remotely Activated Mechanotransduction via Magnetic Nanoparticles Promotes Mineralization Synergistically With Bone Morphogenetic Protein 2: Applications for Injectable Cell Therapy,” Stem cells translational medicine, p. sctm-2014, 2014.
[26]        A. D. Dikina, B. P. Lai, M. Cao, M. Zborowski, and E. Alsberg, “Magnetic field application or mechanical stimulation via magnetic microparticles does not enhance chondrogenesis in mesenchymal stem cell sheets,” Biomaterials science, vol. 5, no. 7, pp. 1241–1245, 2017.
[27]        S. H. Cartmell, J. Dobson, S. B. Verschueren, and A. J. El Haj, “Development of magnetic particle techniques for long-term culture of bone cells with intermittent mechanical activation,” IEEE Transactions on NanoBioscience, vol. 99, no. 2, pp. 92–97, 2002.
[28]        S. Hughes, J. Dobson, and A. J. El Haj, “Magnetic targeting of mechanosensors in bone cells for tissue engineering applications,” Journal of biomechanics, vol. 40, pp. S96–S104, 2007.
[29]        B. Son, H. D. Kim, M. Kim, J. A. Kim, J. Lee, H. Shin, N. S. Hwang, and T. H. Park, “Physical Stimuli‐Induced Chondrogenic Differentiation of Mesenchymal Stem Cells Using Magnetic Nanoparticles,” Advanced healthcare materials, vol. 4, no. 9, pp. 1339–1347, 2015.
[30]        L. Li, K. Y. Mak, J. Shi, C. H. Leung, C. M. Wong, C. W. Leung, C. S. K. Mak, K. Y. Chan, N. M. M. Chan, and E. X. Wu, “Sterilization on dextran-coated iron oxide nanoparticles: Effects of autoclaving, filtration, UV irradiation, and ethanol treatment,” Microelectronic engineering, vol. 111, pp. 310–313, 2013.
[31]        B. Johnstone, T. M. Hering, A. I. Caplan, V. M. Goldberg, and J. U. Yoo, “< i> In Vitro Chondrogenesis of Bone Marrow-Derived Mesenchymal Progenitor Cells,” Experimental cell research, vol. 238, no. 1, pp. 265–272, 1998.
[32]        W. Wang, B. Li, J. Yang, L. Xin, Y. Li, H. Yin, Y. Qi, Y. Jiang, H. Ouyang, and C. Gao, “The restoration of full-thickness cartilage defects with BMSCs and TGF-beta 1 loaded PLGA/fibrin gel constructs,” Biomaterials, vol. 31, no. 34, pp. 8964–8973, 2010.
[33]        T. D. Schmittgen and K. J. Livak, “Analyzing real-time PCR data by the comparative C T method,” Nature protocols, vol. 3, no. 6, p. 1101, 2008.
[34]        G. Yuan, Y. Yuan, K. Xu, and Q. Luo, “Biocompatible PEGylated Fe3O4 nanoparticles as photothermal agents for near-infrared light modulated cancer therapy,” International journal of molecular sciences, vol. 15, no. 10, pp. 18776–18788, 2014.
[35]        M. Jackson, P. H. Watson, W. C. Halliday, and H. H. Mantsch, “Beware of connective tissue proteins: assignment and implications of collagen absorptions in infrared spectra of human tissues,” Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, vol. 1270, no. 1, pp. 1–6, 1995.
[36]        J. Lima, A. I. Gonçalves, M. T. Rodrigues, R. L. Reis, and M. E. Gomes, “The effect of magnetic stimulation on the osteogenic and chondrogenic differentiation of human stem cells derived from the adipose tissue (hASCs),” Journal of Magnetism and Magnetic Materials, vol. 393, pp. 526–536, 2015.
[37]        J. L. Alonso and W. H. Goldmann, “Cellular mechanotransduction,” transport, vol. 1, p. 7, 2016.
[38]        B. C. Low, C. Q. Pan, G. V Shivashankar, A. Bershadsky, M. Sudol, and M. Sheetz, “YAP/TAZ as mechanosensors and mechanotransducers in regulating organ size and tumor growth,” FEBS letters, vol. 588, no. 16, pp. 2663–2670, 2014.
[39]        M. Glogauer, P. Arora, G. Yao, I. Sokholov, J. Ferrier, and C. A. McCulloch, “Calcium ions and tyrosine phosphorylation interact coordinately with actin to regulate cytoprotective responses to stretching,” Journal of Cell Science, vol. 110, no. 1, pp. 11–21, 1997.
[40]        M. Glogauer, J. Ferrier, and C. A. McCulloch, “Magnetic fields applied to collagen-coated ferric oxide beads induce stretch-activated Ca2+ flux in fibroblasts,” American Journal of Physiology-Cell Physiology, vol. 269, no. 5, pp. C1093–C1104, 1995.
[41]        N. J. Sniadecki, “Minireview: a tiny touch: activation of cell signaling pathways with magnetic nanoparticles,” Endocrinology, vol. 151, no. 2, pp. 451–457, 2010.
[42]        K. K. Sethi, V. Mudera, R. Sutterlin, W. Baschong, and R. A. Brown, “Contraction‐mediated pinocytosis of RGD‐peptide by dermal fibroblasts: Inhibition of matrix attachment blocks contraction and disrupts microfilament organisation,” Cytoskeleton, vol. 52, no. 4, pp. 231–241, 2002.
[43]        S. Seetharaman and S. Etienne‐Manneville, “Integrin diversity brings specificity in mechanotransduction,” Biology of the Cell, vol. 110, no. 3, pp. 49–64, 2018.
[44]        D. Fayol, N. Luciani, L. Lartigue, F. Gazeau, and C. Wilhelm, “Managing magnetic nanoparticle aggregation and cellular uptake: a precondition for efficient stem‐cell differentiation and MRI tracking,” Advanced healthcare materials, vol. 2, no. 2, pp. 313–325, 2013.
[45]        Y. Chang, Y. Liu, J. H. Ho, S. Hsu, and O. K. Lee, “Amine‐surface‐modified superparamagnetic iron oxide nanoparticles interfere with differentiation of human mesenchymal stem cells,” Journal of Orthopaedic Research, vol. 30, no. 9, pp. 1499–1506, 2012.
[46]        J.-Y. Su, S.-H. Chen, Y.-P. Chen, and W.-C. Chen, “Evaluation of Magnetic Nanoparticle-Labeled Chondrocytes Cultivated on a Type II Collagen–Chitosan/Poly (Lactic-co-Glycolic) Acid Biphasic Scaffold,” International journal of molecular sciences, vol. 18, no. 1, p. 87, 2017.
[47]        E. Lucchinetti, M. M. Bhargava, and P. A. Torzilli, “The effect of mechanical load on integrin subunits α5 and β1 in chondrocytes from mature and immature cartilage explants,” Cell and tissue research, vol. 315, no. 3, pp. 385–391, 2004.
[48]        T. Kurakawa, K. Kakutani, Y. Morita, Y. Kato, T. Yurube, H. Hirata, S. Miyazaki, Y. Terashima, K. Maeno, and T. Takada, “Functional impact of integrin α5β1 on the homeostasis of intervertebral discs: a study of mechanotransduction pathways using a novel dynamic loading organ culture system,” The Spine Journal, vol. 15, no. 3, pp. 417–426, 2015.
[49]        C. Huang, Y. Charles, K. L. Hagar, L. E. Frost, Y. Sun, and H. S. Cheung, “Effects of cyclic compressive loading on chondrogenesis of rabbit bone‐marrow derived mesenchymal stem cells,” Stem cells, vol. 22, no. 3, pp. 313–323, 2004.
[50]        I. Takahashi, G. H. Nuckolls, K. Takahashi, O. Tanaka, I. Semba, R. Dashner, L. Shum, and H. C. Slavkin, “Compressive force promotes sox9, type II collagen and aggrecan and inhibits IL-1beta expression resulting in chondrogenesis in mouse embryonic limb bud mesenchymal cells,” Journal of cell science, vol. 111, no. 14, pp. 2067–2076, 1998.
[51]        J. F. Schenck, “The role of magnetic susceptibility in magnetic resonance imaging: MRI magnetic compatibility of the first and second kinds,” Medical physics, vol. 23, no. 6, pp. 815–850, 1996.
[52]        H. D. Amin, M. A. Brady, J.-P. St-Pierre, M. M. Stevens, D. R. Overby, and C. R. Ethier, “Stimulation of Chondrogenic Differentiation of Adult Human Bone Marrow-Derived Stromal Cells by a Moderate-Strength Static Magnetic Field,” Tissue Engineering Part A, 2014.
[53]        K. Andreas, R. Georgieva, M. Ladwig, S. Mueller, M. Notter, M. Sittinger, and J. Ringe, “Highly efficient magnetic stem cell labeling with citrate-coated superparamagnetic iron oxide nanoparticles for MRI tracking,” Biomaterials, vol. 33, no. 18, pp. 4515–4525, 2012.
Volume 49, Issue 2
December 2018
Pages 386-394
  • Receive Date: 08 September 2018
  • Revise Date: 10 September 2018
  • Accept Date: 20 September 2018