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Non-viral gene delivery of interleukin-1 receptor antagonist using collagen-hydroxyapatite scaffold protects rat BM-MSCs From IL-1β-mediated inhibition of osteogenesis.

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posted on 19.01.2021, 17:47 by William A Lackington, Maria A Gomez-Sierra, Arlyng Gonzalez Vazquez, Fergal O'Brien, Martin J Stoddart, Keith Thompson
Although most bone fractures typically heal without complications, a small proportion of patients (≤10%) experience delayed healing or potential progression to non-union. Interleukin-1 (IL-1β) plays a crucial role in fracture healing as an early driver of inflammation. However, the effects of IL-1β can impede the healing process if they persist long after the establishment of a fracture hematoma, making it a promising target for novel therapies. Accordingly, the overall objective of this study was to develop a novel gene-based therapy that mitigates the negative effects of IL-1β-driven inflammation while providing a structural template for new bone formation. A collagen-hydroxyapatite scaffold (CHA) was used as a platform for the delivery of nanoparticles composed of pDNA, encoding for IL-1 receptor antagonist (IL-1Ra), complexed to the robust non-viral gene delivery vector, polyethyleneimine (PEI). Utilizing pDNA encoding for Gaussia luciferase and GFP as reporter genes, we found that PEI-pDNA nanoparticles induced a transient gene expression profile in rat bone marrow-derived mesenchymal stromal cells (BM-MSCs), with a transfection efficiency of 14.8 ± 1.8% in 2D. BM-MSC viability was significantly affected by PEI-pDNA nanoparticles as evaluated using CellTiter Blue; however, after 10 days in culture this effect was negligible. Transfection with PEI-pIL-1Ra nanoparticles led to functional IL-1Ra production, capable of antagonizing IL-1β-induced expression of secreted embryonic alkaline phosphatase from HEK-Blue-IL-1β reporter cells. Sustained treatment with IL-1β (0.1, 1, and 10 ng/ml) had a dose-dependent negative effect on BM-MSC osteogenesis, both in terms of gene expression (Alpl and Ibsp) and calcium deposition. BM-MSCs transfected with PEI-IL-1Ra nanoparticles were found to be capable of overcoming the inhibitory effects of sustained IL-1β (1 ng/ml) treatments on in vitro osteogenesis. Ultimately, IL-1Ra gene-activated CHA scaffolds supported mineralization of BM-MSCs under chronic inflammatory conditions in vitro, demonstrating potential for future therapeutic applications in vivo.

Funding

AO Foundation

European Research Council under the European Community’s Horizon 2020 Research and Innovation Programme under ERC grant agreement no. 788753 (ReCaP)

History

Comments

The original article is available at https://www.frontiersin.org/

Published Citation

Lackington WA, Gomez-Sierra MA, González-Vázquez A, O'Brien FJ, Stoddart MJ, Thompson K. Non-viral gene delivery of interleukin-1 receptor antagonist using collagen-hydroxyapatite scaffold protects rat BM-MSCs from IL-1β-mediated inhibition of osteogenesis. Frontiers in Bioengineering and Biotechnology. 2020;8:582012.

Publication Date

6 October 2020

PubMed ID

33123517

Department/Unit

  • Amber (Advanced Material & Bioengineering Research) Centre
  • Anatomy and Regenerative Medicine
  • RCSI Tissue Engineering Group (TERG)

Research Area

  • Biomaterials and Regenerative Medicine

Publisher

Frontiers Media SA

Version

  • Published Version (Version of Record)

Licence

Exports