In Vitro Evaluation of a Nanoparticle-Based mRNA Delivery System for Cells in the Joint
Lisa Sturm,
Bettina Schwemberger,
Ursula Menzel,
Sonja Häckel,
Christoph E. Albers,
Christian Plank,
Jaap Rip,
Mauro Alini,
Andreas Traweger,
Sibylle Grad,
Valentina Basoli
Affiliations
Lisa Sturm
Institute of Tendon and Bone Regeneration, Spinal Cord Injury & Tissue Regeneration Center Salzburg, Paracelsus Medical University, 5020 Salzburg, Austria
Bettina Schwemberger
Institute of Tendon and Bone Regeneration, Spinal Cord Injury & Tissue Regeneration Center Salzburg, Paracelsus Medical University, 5020 Salzburg, Austria
Ursula Menzel
AO Research Institute Davos, 7270 Davos Platz, Switzerland
Sonja Häckel
Department of Orthopaedic Surgery and Traumatology, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
Christoph E. Albers
Department of Orthopaedic Surgery and Traumatology, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
Christian Plank
ETHRIS GmbH, 82152 Planegg, Germany
Jaap Rip
20Med Therapeutics B.V., Galileiweg 8, 2333BD Leiden, The Netherlands
Mauro Alini
AO Research Institute Davos, 7270 Davos Platz, Switzerland
Andreas Traweger
Institute of Tendon and Bone Regeneration, Spinal Cord Injury & Tissue Regeneration Center Salzburg, Paracelsus Medical University, 5020 Salzburg, Austria
Sibylle Grad
AO Research Institute Davos, 7270 Davos Platz, Switzerland
Valentina Basoli
AO Research Institute Davos, 7270 Davos Platz, Switzerland
Biodegradable and bioresponsive polymer-based nanoparticles (NPs) can be used for oligonucleotide delivery, making them a promising candidate for mRNA-based therapeutics. In this study, we evaluated and optimized the efficiency of a cationic, hyperbranched poly(amidoamine)s-based nanoparticle system to deliver tdTomato mRNA to primary human bone marrow stromal cells (hBMSC), human synovial derived stem cells (hSDSC), bovine chondrocytes (bCH), and rat tendon derived stem/progenitor cells (rTDSPC). Transfection efficiencies varied among the cell types tested (bCH 28.4% ± 22.87, rTDSPC 18.13% ± 12.07, hBMSC 18.23% ± 14.80, hSDSC 26.63% ± 8.81) and while an increase of NPs with a constant amount of mRNA generally improved the transfection efficiency, an increase of the mRNA loading ratio (2:50, 4:50, or 6:50 w/w mRNA:NPs) had no impact. However, metabolic activity of bCHs and rTDSPCs was significantly reduced when using higher amounts of NPs, indicating a dose-dependent cytotoxic response. Finally, we demonstrate the feasibility of transfecting extracellular matrix-rich 3D cell culture constructs using the nanoparticle system, making it a promising transfection strategy for musculoskeletal tissues that exhibit a complex, dense extracellular matrix.
