Stimulation of 3D osteogenesis by mesenchymal stem cells using a nanovibrational bioreactor

Abstract

Bone grafts are one of the most commonly transplanted tissues. However, autologous grafts are in short supply, and can be associated with pain and donor-site morbidity. The creation of tissue-engineered bone grafts could help to fulfil clinical demand and provide a crucial resource for drug screening. Here, we show that vibrations of nanoscale amplitude provided by a newly developed bioreactor can differentiate a potential autologous cell source, mesenchymal stem cells (MSCs), into mineralized tissue in 3D. We demonstrate that nanoscale mechanotransduction can stimulate osteogenesis independently of other environmental factors, such as matrix rigidity. We show this by generating mineralized matrix from MSCs seeded in collagen gels with stiffness an order of magnitude below the stiffness of gels needed to induce bone formation in vitro. Our approach is scalable and can be compatible with 3D scaffolds.

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Fig. 1: Development of the nanovibrational bioreactor.
Fig. 2: Characterization of 3D collagen gel culture on the vibrational bioreactor.
Fig. 3: Confirmation of bone mineralization in 3D through nanovibrational stimulation.
Fig. 4: Testing for channel and phenotype sensitivity to cytoskeletal tension.
Fig. 5: Testing for TRPV involvement in nanovibration-stimulated osteogenic pathways.
Fig. 6: Testing for TRPV and Wnt involvement in nanovibration-stimulated osteogenic stimulation using metabolomics.
Fig. 7: Testing for TRPV involvement in nanovibration-stimulated osteogenesis.

Change history

  • 22 November 2017

    In the version of this Article originally published, in Fig. 4f, the asterisk was missing; in Fig. 6a–c, the labels ‘Wnt/β-catenin signalling’, ‘Wnt/Ca+ pathway’ and ‘ERK’ and their associated lines/arrows were missing; and in Fig. 6d and in the sentence beginning “In MSCs that were...”, ‘myosin’ and ‘nanostimulated’, respectively, were spelt incorrectly. These errors have now been corrected in all versions of the Article.

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Acknowledgements

This work was supported by grants to S.R. and M.J.D. from the BBSRC, BBSRC/SFI and EPSRC (BB/N012690/1, BB/P00220X/1, EP/N013905/1, EP/N012631/1 and EP/P001114/1), along with a Wolfson Merit Award from The Royal Society and a programme grant from Find a Better Way. M.J.P.B. is funded by SFI grant nos. 11/SIRG/B2135 and 13/RC/2073. P.G.C. was funded by an STFC/BBSRC fellowship. We thank J. Hough, H. Nikukar, I. Tifenbrun and K. Robertson for their discussion, C.-A. Smith for technical support, and E. Manson for help with metabolite analysis. We also thank C. Boyle, S. Robertson and P. Campsie for help with the construction of the bioreactor.

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A.S.G.C. was the inspiration behind this work, to whom it is dedicated. P.M.T., P.G.C., G.D.P., J.Y., V.J., W.O., C.G.-G., D.T. and C.V.G. performed the laboratory experiments. P.M.T., P.G.C., G.D.P., W.O., K.B., G.B., M.J.P.B., M.S.-S., S.R. and M.J.D. analysed the data. P.M.T., P.G.C., G.D.P., M.J.P.B., A.S.G.C., M.S.S., S.R. and M.J.D. devised experiments. S.R. and M.J.D. supervised the research. P.M.T., P.G.C., G.D.P., M.S.S., S.R. and M.J.D. wrote the manuscript. P.M.T., P.G.C., S.R. and M.J.D. revised the manuscript.

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Correspondence to Stuart Reid or Matthew J. Dalby.

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Tsimbouri, P.M., Childs, P.G., Pemberton, G.D. et al. Stimulation of 3D osteogenesis by mesenchymal stem cells using a nanovibrational bioreactor. Nat Biomed Eng 1, 758–770 (2017). https://doi.org/10.1038/s41551-017-0127-4

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