| Autor: |
Muenzebrock KA; Department of Orthopedics, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands., Ho FYW; Department of Orthopedics, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands., Pontes AP; 20Med Therapeutics BV, 2333 BD Leiden, The Netherlands., Jorquera-Cordero C; Department of Chemistry and Pharmaceutical Sciences, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands., Utomo L; Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CS Utrecht, The Netherlands., Garcia JP; Department of Orthopedics, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands., Willems PC; Department of Orthopedic Surgery, Maastricht University Medical Center, 6229 HX Maastricht, The Netherlands., Welting TJM; Department of Orthopedic Surgery, Maastricht University Medical Center, 6229 HX Maastricht, The Netherlands., Rip J; 20Med Therapeutics BV, 2333 BD Leiden, The Netherlands., Creemers LB; Department of Orthopedics, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands. |
| Abstrakt: |
Chronic lower back pain caused by intervertebral disc degeneration and osteoarthritis (OA) are highly prevalent chronic diseases. Although pain management and surgery can alleviate symptoms, no disease-modifying treatments are available. mRNA delivery could halt inflammation and degeneration and induce regeneration by overexpressing anti-inflammatory cytokines or growth factors involved in cartilage regeneration. Here, we investigated poly(amidoamine)-based polymeric nanoparticles to deliver mRNA to human joint and intervertebral disc cells. Human OA chondrocytes, human nucleus pulposus (NP) cells, human annulus fibrosus (AF) cells, fibroblast-like synoviocytes (FLS) and M1-like macrophages were cultured and transfected with uncoated or PGA-PEG-coated nanoparticles loaded with EGFP-encoding mRNA. Cell viability and transfection efficiency were analyzed for all cell types. Nanoparticle internalization was investigated in FLS and M1-like macrophages. No significant decrease in cell viability was observed in most conditions. Only macrophages showed a dose-dependent reduction of viability. Transfection with either nanoparticle version resulted in EGFP expression in NP cells, AF cells, OA chondrocytes and FLS. Macrophages showed internalization of nanoparticles by particle-cell co-localization, but no detectable expression of EGFP. Taken together, our data show that poly (amidoamine)-based nanoparticles can be used for mRNA delivery into cells of the human joint and intervertebral disc, indicating its potential future use as an mRNA delivery system in OA and IVDD, except for macrophages. |
