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Journal of orthopaedic research : official publication of the Orthopaedic Research Society2019; 37(6); 1310-1317; doi: 10.1002/jor.24209

Genetic modification of scAAV-equine-BMP-2 transduced bone-marrow-derived mesenchymal stem cells before and after cryopreservation: An “off-the-shelf” option for fracture repair.

Abstract: Optimizing the environment of complex bone healing and improving treatment of catastrophic bone fractures and segmental bone defects remains an unmet clinical need both human and equine veterinary medical orthopaedics. The objective of this study was to determine whether scAAV-equine-BMP-2 transduced cells would induce osteogenesis in equine bone marrow derived mesenchymal stem cells (BMDMSCs) in vitro, and if these cells could be cryopreserved in an effort to osteogenically prime them as an "off-the-shelf" gene therapeutic approach for fracture repair. Our study found that transgene expression is altered by cell expansion, as would be expected by a transduction resulting in episomal transgene expression, and that osteoinductive levels could still be achieved 5 days after recovery, and protein expression would continue up to 14 days after transduction. This is the first evidence that cryopreservation of genetically modified BMDMSCs would not alter the osteoinductive potential or clinical use of allogeneic donor cells in cases of equine fracture repair. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1310-1317, 2019.
© 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
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Publication Date: 2019-02-21 PubMed ID: 30578639PubMed Central: PMC8366205DOI: 10.1002/jor.24209Google Scholar: Lookup
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  • N.I.H.
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research studied the potential of genetically-modified, bone marrow derived mesenchymal stem cells for use in bone fracture repair for humans and horses, finding that cryopreservation did not affect the cells’ ability to promote bone growth.

Introduction to the Research Topic

  • The study sought to improve the treatment of complex bone fractures and segmental bone defects, which remain challenging for both human and equine orthopaedics.
  • The researchers tried to induce bone growth in horse bone marrow derived mesenchymal stem cells (BMDMSCs), a type of cells that can develop into bone, using genetically transduced cells with a gene called scAAV-equine-BMP-2.

Cryopreservation and Genetic Modification

  • This study also investigated whether these genetically modified cells could be effectively cryopreserved, i.e., frozen for storage and later use.
  • If successful, this could lead to a readily available “off-the-shelf” gene therapeutic approach for fracture repair.

Key Findings

  • The results demonstrated that the gene expression in the genetically modified BMDMSCs was altered when the cells were multiplied or expanded. This is typical of cells that have undergone gene transduction, a process that brings foreign DNA into cells.
  • However, the researchers discovered that even after cell expansion, bone growth-promoting (osteoinductive) levels could still be achieved in the cells 5 days after being thawed from cryopreservation, and protein expression continued for up to 14 days after gene transduction.

Significance of the Study

  • This is the first study to show that cryopreservation of genetically modified BMDMSCs does not have any significant impact on their osteoinductive potential or their ability to be used clinically in fracture repairs.
  • The successful preservation of these cells means that they could be made readily available as an “off-the-shelf” treatment option for fractures, potentially leading to more effective and quicker recovery for human and equine patients.

Cite This Article

APA
Ball AN, Phillips JN, McIlwraith CW, Kawcak CE, Samulski RJ, Goodrich LR. (2019). Genetic modification of scAAV-equine-BMP-2 transduced bone-marrow-derived mesenchymal stem cells before and after cryopreservation: An “off-the-shelf” option for fracture repair. J Orthop Res, 37(6), 1310-1317. https://doi.org/10.1002/jor.24209

Publication

Journal of orthopaedic research : official publication of the Orthopaedic Research Society
ISSN: 1554-527X
NlmUniqueID: 8404726
Country: United States
Language: English
Volume: 37
Issue: 6
Pages: 1310-1317

Researcher Affiliations

Ball, Alyssa N
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Phillips, Jennifer N
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
McIlwraith, C Wayne
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Kawcak, Christopher E
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Samulski, Richard J
  • Gene Therapy Center, University of North Carolina, Chapel Hill, NC 27599, USA.
Goodrich, Laurie R
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523, USA.

MeSH Terms

  • Animals
  • Bone Morphogenetic Protein 2 / genetics
  • Cryopreservation
  • Dependovirus / genetics
  • Fracture Healing
  • Genetic Therapy / methods
  • Horses
  • Transduction, Genetic

Grant Funding

  • T32 OD012201 / NIH HHS

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Citations

This article has been cited 8 times.
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