Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts

International Journal of Nanomedicine, Jan 2015

Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts Antonella Rocca,1,2 Attilio Marino,1,2 Veronica Rocca,3 Stefania Moscato,4 Giuseppe de Vito,5,6 Vincenzo Piazza,5 Barbara Mazzolai,1 Virgilio Mattoli,1 Thu Jennifer Ngo-Anh,7 Gianni Ciofani1 1Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, Pontedera, Italy, 2Scuola Superiore Sant’Anna, The BioRobotics Institute, Pontedera, Italy, 3Università di Pisa, Dipartimento di Ingegneria dell’Informazione, Pisa, Italy, 4Università di Pisa, Dipartimento di Medicina Clinica e Sperimentale, Pisa, Italy, 5Istituto Italiano di Tecnologia, Center for Nanotechnology Innovation @NEST, Pisa, Italy, 6Scuola Normale Superiore, NEST, Pisa, Italy, 7Directorate of Human Spaceflight and Operations, European Space Agency, Noordwijk, the Netherlands Background: Enhancement of the osteogenic potential of mesenchymal stem cells (MSCs) is highly desirable in the field of bone regeneration. This paper proposes a new approach for the improvement of osteogenesis combining hypergravity with osteoinductive nanoparticles (NPs).Materials and methods: In this study, we aimed to investigate the combined effects of hypergravity and barium titanate NPs (BTNPs) on the osteogenic differentiation of rat MSCs, and the hypergravity effects on NP internalization. To obtain the hypergravity condition, we used a large-diameter centrifuge in the presence of a BTNP-doped culture medium. We analyzed cell morphology and NP internalization with immunofluorescent staining and coherent anti-Stokes Raman scattering, respectively. Moreover, cell differentiation was evaluated both at the gene level with quantitative real-time reverse-transcription polymerase chain reaction and at the protein level with Western blotting.Results: Following a 20 g treatment, we found alterations in cytoskeleton conformation, cellular shape and morphology, as well as a significant increment of expression of osteoblastic markers both at the gene and protein levels, jointly pointing to a substantial increment of NP uptake. Taken together, our findings suggest a synergistic effect of hypergravity and BTNPs in the enhancement of the osteogenic differentiation of MSCs.Conclusion: The obtained results could become useful in the design of new approaches in bone-tissue engineering, as well as for in vitro drug-delivery strategies where an increment of nanocarrier internalization could result in a higher drug uptake by cell and/or tissue constructs. Keywords: mesenchymal stem cells, hypergravity, barium titanate nanoparticles, osteogenesis

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Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts

International Journal of Nanomedicine Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts attilio Marino 1 2 3 8 Vincenzo Piazza 3 6 8 Barbara Mazzolai 2 3 8 Virgilio Mattoli 2 3 8 Thu Jennifer Ngo-anh 3 4 8 0 Università di Pisa , Dipartimento di Ingegneria dell'Informazione, Pisa , Italy 1 s cuola s uperiore s ant'a nna, The Bior obotics Institute , Pontedera , Italy 2 Istituto Italiano di Tecnologia, center for Micro-Bio robotics @ sssa , Pontedera , Italy 3 giuseppe de Vito 4 Directorate of h uman s paceflight and Operations , european s pace agency, Noordwijk , the Netherlands 5 s cuola Normale superiore, N esT , Pisa , Italy 6 Istituto Italiano di Tecnologia, c enter for Nanotechnology Innovation @NesT , Pisa , Italy 7 Università di Pisa, Dipartimento di Medicina clinica e sperimentale , Pisa , Italy 8 Pump @ 806 nm Stokes @ 1 , 000 nm to 1,100 nm 8 1 0 2 - l u J - 2 1 n o 7 0 2 . 6 4 . 9 5 . 7 3 y b / m o c . s s e r p e v PowerdbyTCPDF(ww.tcpdf.org) O r I g I N a l r e s e a r c h Veronica rocca 3 stefania Moscato 4 gianni ciofani 1 Background: Enhancement of the osteogenic potential of mesenchymal stem cells (MSCs) is highly desirable in the field of bone regeneration. This paper proposes a new approach for the improvement of osteogenesis combining hypergravity with osteoinductive nanoparticles Materials and methods: In this study, we aimed to investigate the combined effects of hypergravity and barium titanate NPs (BTNPs) on the osteogenic differentiation of rat MSCs, and the hypergravity effects on NP internalization. To obtain the hypergravity condition, we used a large-diameter centrifuge in the presence of a BTNP-doped culture medium. We analyzed cell morphology and NP internalization with immunofluorescent staining and coherent anti-Stokes Raman scattering, respectively. Moreover, cell differentiation was evaluated both at the gene level with quantitative real-time reverse-transcription polymerase chain reaction and at the protein level with Western blotting. Results: Following a 20 g treatment, we found alterations in cytoskeleton conformation, cellular shape and morphology, as well as a significant increment of expression of osteoblastic markers both at the gene and protein levels, jointly pointing to a substantial increment of NP uptake. Taken together, our findings suggest a synergistic effect of hypergravity and BTNPs in the enhancement of the osteogenic differentiation of MSCs. Conclusion: The obtained results could become useful in the design of new approaches in bone-tissue engineering, as well as for in vitro drug-delivery strategies where an increment of nanocarrier internalization could result in a higher drug uptake by cell and/or tissue constructs. mesenchymal stem cells; hypergravity; barium titanate nanoparticles; osteogenesis - open access to scientific and medical research Introduction Physical stimuli can significantly alter cellular behavior, giving rise to biochemical signals involved in molecular response.1 This process is called mechanotransduction, and the responsible structures sensitive to mechanical forces are most probably cytoskeleton elements.2 A number of studies have demonstrated that cells are sensitive to several kinds of physical cues (shear stress, topography, mechanical deformation, etc), influencing cell migration,3 differentiation,4 and proliferation.5 Among these stimuli, gravity is required for the correct development of land-based organisms, and in particular for the skeleton and for the muscle and nervous systems.6 An increasing amount of research is focused on the effects of gravity alterations on the physiological processes, but also on the possibility to exploit this stimulus as a potential therapeutic cue.7–9 As an example, improved regeneration of infarcted myocardium has been achieved after injection of stem cells differentiated following a 2 g hypergravity treatment, which enhanced the activities of cardiac marker MEF-2 by promoting the nuclear export of histone deacetylase 5.10 Chang et al investigated altered gravity effects on human lung adenocarcinoma, demonstrating the ability of simulated microgravity to decrease the metastatic potential of this tumor cell line.11 Other researchers used microgravity stimulation as an approach for the development of a large amount of β-cell spheroids, which once transplanted in mice are able to improve the symptoms of diabetes.12 Among different tissues, bone is particularly affected by altered gravity conditions: evidence regarding bone regeneration suggests that hypergravity exposure – conversely to microgravity, which negatively affects osteogenesis – may enhance the osteogenic potential of osteoblast precursors.13 The ability of mesenchymal stem cells (MSCs) to differentiate into osteoblasts is well known, but the osteogenic potential of MSCs decreases with the prolonged culture duration necessary to obtain an appropriat (...truncated)


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Antonella Rocca, Attilio Marino, Veronica Rocca, Stefania Moscato, Giuseppe de Vito, Vincenzo Piazza, Barbara Mazzolai, Virgilio Mattoli, Thu Jennifer Ngo-Anh, Gianni Ciofani. Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts, International Journal of Nanomedicine, 2015, pp. 433-445, DOI: 10.2147/IJN.S76329