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Biomimetics and Stem Cells pp 149–160Cite as

The Delivery and Evaluation of RNAi Therapeutics for Heterotopic Ossification Pathologies

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1202))

Abstract

RNA interference (RNAi) is a powerful tool being used to develop therapies for pathologies caused by gene overexpression. Heterotopic ossification pathologies such as trauma-induced heterotopic ossification and fibrodysplasia ossificans progressiva may be treatable with an RNAi approach. However, there is a lack of consensus in literature regarding the delivery conditions and evaluation of RNAi therapeutics in these disease models. Here, we describe in vitro protocols for the delivery of polymer-based RNAi therapeutics as well as a streamlined strategy for the assessment of osteoblast lineage progression due to dysregulated bone morphogenetic protein signaling. This strategy focuses on the quantification of early-stage osteoblast transcription factors RUNX2 and OSX, followed by the measurement of alkaline phosphatase activity and late-stage matrix deposition.

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Acknowledgement

This work was supported by Department of Defense grant W81XWH-11-2-0073, through the Defense Medical Research and Development Program.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA

    Arun R. Shrivats

  2. Departments of Biomedical Engineering and Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA

    Jeffrey O. Hollinger

Authors
  1. Arun R. Shrivats
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  2. Jeffrey O. Hollinger
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Corresponding author

Correspondence to Jeffrey O. Hollinger .

Editor information

Editors and Affiliations

  1. Department of Biomedical Engineering and Department of Medicine, Columbia University, New York, New York, USA

    Gordana Vunjak-Novakovic

  2. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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© 2013 Springer Science+Business Media New York

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Shrivats, A.R., Hollinger, J.O. (2013). The Delivery and Evaluation of RNAi Therapeutics for Heterotopic Ossification Pathologies. In: Vunjak-Novakovic, G., Turksen, K. (eds) Biomimetics and Stem Cells. Methods in Molecular Biology, vol 1202. Humana Press, New York, NY. https://doi.org/10.1007/7651_2013_34

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  • DOI: https://doi.org/10.1007/7651_2013_34

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1331-2

  • Online ISBN: 978-1-4939-1332-9

  • eBook Packages: Springer Protocols

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