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A Comparative Study of the Bone Regenerative Effect of Chemically Modified RNA Encoding BMP-2 or BMP-9

  • Research Article
  • Theme: Recent Advances in Musculoskeletal Tissue Engineering
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Abstract

Employing cost-effective biomaterials to deliver chemically modified ribonucleic acid (cmRNA) in a controlled manner addresses the high cost, safety concerns, and lower transfection efficiency that exist with protein and gene therapeutic approaches. By eliminating the need for nuclear entry, cmRNA therapeutics can potentially overcome the lower transfection efficiencies associated with non-viral gene delivery systems. Here, we investigated the osteogenic potential of cmRNA-encoding BMP-9, in comparison to cmRNA-encoding BMP-2. Polyethylenimine (PEI) was used as a vector to increase in vitro transfection efficacy. Complexes of PEI-cmRNA (encoding BMP-2 or BMP-9) were fabricated at an amine (N) to phosphate (P) ratio of 10 and characterized for transfection efficacy in vitro using human bone marrow stromal cells (BMSCs). The osteogenic potential of BMSCs treated with these complexes was determined by evaluating the expression of bone-specific genes as well as through the detection of bone matrix deposition. It was found that alkaline phosphatase (ALP) expression 3 days post transfection in the group treated with BMP-9-cmRNA was significantly higher than that in the group that received BMP-2-cmRNA treatment. Alizarin red staining and atomic absorption spectroscopy demonstrated enhanced osteogenic differentiation as evidenced by increased bone matrix production by the BMSCs treated with BMP-9-cmRNA when compared to cells treated with BMP-2-cmRNA. In vivo studies showed increased bone formation in calvarial defects treated with the BMP-9-cmRNA and BMP-2-cmRNA collagen scaffolds when compared to empty defects. The connectivity density of the regenerated bone was higher (2-fold-higher) in the group that received BMP-9-cmRNA compared to BMP-2-cmRNA. Together, these findings suggest that cmRNA-activated matrix encoding osteogenic molecules can provide a powerful strategy for bone regeneration with significant clinical translational potential.

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Acknowledgements

This study was supported by an NIH R21 grant (1R21DE024206-01A1), the University of Iowa Start-up Grant, the ITI Foundation for the Promotion of Implantology, Switzerland (ITI Research Grant No. 855 2012), the Osteology Foundation Grant (12–054), the Sunstar—American Academy of Periodontology Foundation Research Fellowship, and the Lyle and Sharon Bighley Professorship. The authors would like to thank Ryan D. Ross and Rick Sumner for their assistance in obtaining MicroCT images. Rush University Medical Center MicroCT/Histology Core resources were used. We also would like to thank Chantal Allamargot for technical support with histological image acquisition. Imaging equipment at the University of Iowa Core Microscopy Research Facility was used.

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Authors and Affiliations

  1. Division of Pharmaceutics and Translational Therapeutics, University of Iowa College of Pharmacy, Iowa City, Iowa, USA

    Behnoush Khorsand & Aliasger K. Salem

  2. Department of Periodontics, University of Iowa College of Dentistry, Iowa City, Iowa, USA

    Satheesh Elangovan & Aliasger K. Salem

  3. Department of Prosthodontics, University of Iowa College of Dentistry, Iowa City, Iowa, USA

    Liu Hong

  4. Department of Pediatrics (Section I), Translational Genomics and Gene Therapy, University of Tübingen, Wilhelmstr. 27, 72074, Tübingen, Germany

    Alexander Dewerth & Michael S. D. Kormann

Authors
  1. Behnoush Khorsand
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  2. Satheesh Elangovan
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  3. Liu Hong
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  4. Alexander Dewerth
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  5. Michael S. D. Kormann
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  6. Aliasger K. Salem
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Corresponding authors

Correspondence to Satheesh Elangovan or Aliasger K. Salem.

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Guest Editor: Aliasger K. Salem

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Khorsand, B., Elangovan, S., Hong, L. et al. A Comparative Study of the Bone Regenerative Effect of Chemically Modified RNA Encoding BMP-2 or BMP-9. AAPS J 19, 438–446 (2017). https://doi.org/10.1208/s12248-016-0034-8

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  • DOI: https://doi.org/10.1208/s12248-016-0034-8

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