Wnt-modified materials mediate asymmetric stem cell division to direct human osteogenic tissue formation for bone repair

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Abstract

The maintenance of human skeletal stem cells (hSSCs) and their progeny in bone defects is a major challenge. Here, we report on a transplantable bandage containing a three-dimensional Wnt-induced osteogenic tissue model (WIOTM). This bandage facilitates the long-term viability of hSSCs (8 weeks) and their progeny, and enables bone repair in an in vivo mouse model of critical-sized calvarial defects. The newly forming bone is structurally comparable to mature cortical bone and consists of human and murine cells. Furthermore, we show that the mechanism of WIOTM formation is governed by Wnt-mediated asymmetric cell division of hSSCs. Covalently immobilizing Wnts onto synthetic materials can polarize single dividing hSSCs, orient the spindle and simultaneously generate a Wnt-proximal hSSC and a differentiation-prone Wnt-distal cell. Our results provide insight into the regulation of human osteogenesis and represent a promising approach to deliver human osteogenic constructs that can survive in vivo and contribute to bone repair.

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Fig. 1: Localized Wnt3a orients division in hSSCs and induces asymmetric distribution of Wnt/β-catenin pathway components.
Fig. 2: Localized Wnt3a induces asymmetric distribution of cell fate markers in dividing cells and the WIOTM.
Fig. 3: In vitro functional characterization of the Wnt3a-bandage.
Fig. 4: In vivo functional evaluation of Wnt3a- and WIOTM-bandages in the repair of critical-sized bone defects.
Fig. 5: In vivo characterization of bone formation driven by Wnt3a- and WIOTM-bandages.
Fig. 6: In vivo characterization of connective-like tissue of bone defects treated with the bandages.

Data availability

All numerical data included in the production of figures, plots and statistical analysis can be found in the figures, main text and Source Data. Additional statistical analysis values (F values, t values, degrees of freedom and P values) are presented in the Source Data documents. Source data are provided with this paper.

Change history

  • 16 October 2020

    In the version of this Article originally published, the links to the Supplementary Information and Source Data files were jumbled; these have now been corrected.

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Acknowledgements

We thank A. Streit, E. Gentleman, A. Grants and H. Drukmann for discussions about the project. We thank P. Sharpe, T. Desai, N. Ali and M. Tewary for critical reading of the manuscript. We thank C. Healy for his help with µCT imaging. We acknowledge financial support from the Department of Health through the National Institute for Health Research (NIHR) Comprehensive Biomedical Research Centre award to the Guy’s & St Thomas’ National Health Service Foundation Trust in partnership with King’s College London and the King’s College Hospital NHS Foundation Trust. K.J.L. and D.H.D. received funding from CAPES, Dental Institute Seed Funding and the BBSRC. This project was supported mainly by a Sir Henry Dale Fellowship (102513/Z/13/Z, S.J.H.). Additional financial support from the UK Regenerative Medicine Platform (MR/R015635/1, S.J.H.) and a London Advanced Therapy Award (S.J.H.) is also acknowledged.

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Contributions

S.J.H. conceived the project and designed the experiments. J.R., Y.O., S.S.N., S.J. and S.J.H. performed and analysed the in vitro experiments. Y.O. and D.H.D. performed the in vivo experiments. Y.O. and J.R. analysed the µCT data. J.R., S.S.N. and S.J. performed the histological analysis. J.R. and S.J.H. interpreted the data. A.J.E.H. and K.J.L. commented on the in vivo experiments. J.R., S.J. and S.J.H. prepared the figures. S.J.H. wrote the manuscript. All authors commented on the manuscript. S.J.H. led the project and provided financial support for the project.

Corresponding author

Correspondence to Shukry J. Habib.

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Competing interests

King’s College London has filed a patent application (P6125GB00) on Tissue Regeneration Patch that includes the Wnt-bandage and WIOTM-bandage.

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Okuchi, Y., Reeves, J., Ng, S.S. et al. Wnt-modified materials mediate asymmetric stem cell division to direct human osteogenic tissue formation for bone repair. Nat. Mater. (2020). https://doi.org/10.1038/s41563-020-0786-5

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