Osteogenic Potential Reduction in Mesenchymal Stem Cells under Prolonged Simulated Microgravity
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Osteogenic Potential Reduction in Mesenchymal Stem Cells under Prolonged Simulated Microgravity

A.Y. Ratushnyy*, O.V. Grigorieva, L.B. Buravkova

Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, 123007 Russia

E-mail: *Ratushkin@mail.ru

Received January 30, 2017

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Abstract

Increasing the duration of orbital space flights up to 6–12 months and planning interplanetary missions actualizes the need for a better understanding of the mechanisms of osteopenia caused by microgravity. Investigation of mesenchymal stem cells (MSCs) that support the tissue homeostasis under microgravity conditions allows a deeper insight into the processes underlying bone loss. The purpose of this study was to investigate the osteogenic potential of MSCs under prolonged simulated microgravity by clinorotation. Using the method of mineralized matrix detection, it has been found that MSCs osteogenic potential decreased after long-term clinorotation. The investigation of major osteogenic gene expression has showed decreased trans­criptional activity in RUNX2, ALPL-1, Col-1, but increased expression of PPARγ. One of the reasons for the decreased osteogenic potential of MSCs may be an increased level of reactive oxygen species (ROS) after 30 days of clinorotation. ROS may affect cellular signaling cascades, such as Wnt, Hedgehog and FOXO pathways, thereby leading to a shift of the differentiation potential to adipogenesis.

Keywords: mesenchymal stem cells (MSCs), simulated microgravity, osteogenic differentiation, reactive oxygen species (ROS)

Acknowledgments. The study was supported by the project no. NSh-7479.2016.4.

Figure Captions

Fig. 1. Osteogenic differentiation, alizarin red staining of mineralized matrix, light microscopy. Magnification: 100×.

Fig. 2. Quantitative evaluation of the mineralization level of extracellular MSC matrix by measuring the optical density of cohered color material (alizarin red) dissolved in DMSO.

Fig. 3. Differential expression of differentiation genes. Data are presented as mean values ? standard deviations. Asterisk indicates significant differences (p ≤ 0.05) from control.

Fig. 4. Fluorescence intensity of H2DCFDA showing the level of reactive oxygen species in cells.

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For citation: Ratushnyy A.Y., Grigorieva O.V., Buravkova L.B. Osteogenic potential reduction in mesenchymal stem cells under prolonged simulated microgravity. Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki, 2017, vol. 159, no. 2, pp. 206–216. (In Russian)


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