Abstract
Tendon and ligament injuries are significant contributors to musculoskeletal injuries. Unfortunately, traditional methods of repair are not uniformly successful and can require revision surgery. Our research is focused on identifying appropriate animal injury models and using tissue-engineered constructs (TECs) from bone-marrow-derived mesenchymal stem cells and collagen scaffolds. Critical to this effort has been the development of functional tissue engineering (FTE). We first determine the in vivo mechanical environment acting on the tissue and then precondition the TECs in culture with aspects of these mechanical signals to improve repair outcome significantly. We describe here a detailed protocol for conducting several complete iterations around our FTE 'road map.' The in vitro portion, from bone marrow harvest to TEC collection, takes 54 d. The in vivo portion, from TEC implantation to limb harvest, takes 84 d. One complete loop around the tissue engineering road map, as presented here, takes 138 d to complete.
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Acknowledgements
This research was supported by NIH Grants AR46574, AR56943 and EB002361. We acknowledge the assistance of Gino Bradica, Shun Yoshida, Tatiana Mavridis, Eric Schantz, Rita Angel, Angela M Sklenka, Heather Powell, Hani Awad, Matthew Dressler and Abhishek Jain.
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All the authors contributed extensively to the protocol described in this paper. D.L.B. and J.T.S. designed experiments, developed and refined the protocol, supervised data analysis and wrote the paper. C.G. gave technical support for cell culture, developed and refined the protocol and wrote the paper. K.R.C.K. designed and performed experiments, refined the protocol, analyzed data and wrote the paper. G.P.B. and M.T.G. designed experiments, performed surgical procedures and edited the paper. N.A.D. designed experiments, refined the protocol and wrote the paper. N.J.-M. and V.S.N. designed and performed experiments, developed and refined the protocol, analyzed data and edited the paper. All the authors discussed the results and implications of the findings.
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Butler, D., Gooch, C., Kinneberg, K. et al. The use of mesenchymal stem cells in collagen-based scaffolds for tissue-engineered repair of tendons. Nat Protoc 5, 849–863 (2010). https://doi.org/10.1038/nprot.2010.14
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DOI: https://doi.org/10.1038/nprot.2010.14
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