Modulation of osteoarthritis-related microRNAs using locked nucleic acid-antisense oligonucleotides boosts chondrogenic lineage commitment of mesenchymal progenitors
BACKGROUND Osteoarthritis (OA) is marked by progressive cartilage degeneration and limited intrinsic repair. Mesenchymal stem cells (MSCs) show promise for cartilage regeneration, yet unstable chondrogenesis and a tendency towards hypertrophy constrain their efficacy. Dysregulated osteoarthritic mic...
| Autores: | , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2026 |
| País: | España |
| Institución: | Revista Iberoamericana de Educación Musical (RIEM) |
| Repositorio: | UCrea Repositorio Abierto de la Universidad de Cantabria |
| Idioma: | inglés |
| OAI Identifier: | oai:dnet:ucreareposit::7f4563aca3aa4f8bf5ae10cc95ce7bc4 |
| Acceso en línea: | https://hdl.handle.net/10902/40175 |
| Access Level: | acceso abierto |
| Palabra clave: | Osteoarthritis Mesenchymal stem cells Chondrogenesis MicroRNA Locked nucleic acid-antisense oligonucleotides Cartilage regeneration Micromass culture |
| Sumario: | BACKGROUND Osteoarthritis (OA) is marked by progressive cartilage degeneration and limited intrinsic repair. Mesenchymal stem cells (MSCs) show promise for cartilage regeneration, yet unstable chondrogenesis and a tendency towards hypertrophy constrain their efficacy. Dysregulated osteoarthritic microRNAs (miRNAs), including miR-16-5p, miR-30b-5p, miR-146a-5p, and miR-193b-3p, negatively influence chondrogenic programmes and matrix homeostasis. We hypothesized that ex vivo priming of MSCs with locked nucleic acid-antisense oligonucleotides (LNA-ASOs) targeting these anti-chondrogenic miRNAs would stabilize early chondrogenesis and enhance matrix formation. AIM To determine whether transient ex vivo LNA-ASO silencing of OA-related miRNAs could improve the chondrogenesis of MSCs. METHODS Rat bone-marrow MSCs were transfected with LNA-ASOs against miR-16-5p, miR-30b-5p, miR-146a-5p, and miR-193b-3p (single or combined) and induced into high-density micromass cultures. On day 11, quantitative real-time polymerase chain reaction assessed Sox9, Acan, Runx2, Mef2C. On day 21, histology (Safranin O, Alcian Blue/Fast Red, Masson?s trichrome, Picrosirius Red under polarised light) and DMMB (sulphated glycosaminoglycans normalised to dsDNA) quantified cartilage-like matrix. Transfection efficiency was measured by flow cytometry/confocal microscopy. Statistics compared each condition with a power inhibitor control. RESULTS All LNA-ASOs achieved transfection efficiencies greater than 85%. The combination miR-30b-5p + miR-193b-3p produced the most favourable molecular profile at day 11, with higher Sox9 and Acan expression and lower Runx2 compared with the power inhibitor control, outperforming single-inhibitor conditions. miR-30b-5p alone and miR-30b-5p + miR-146a-5p also improved chondrogenic markers, whereas the miR-16-5p/miR-30b-5p combination showed a pattern prone to hypertrophy. At day 21, inhibition of miR-30b-5p/miR-193b-3p yielded a stronger Safranin O and Alcian Blue staining and more abundant, better-organized collagen on Masson?s trichrome and Picrosirius Red. DMMB analysis (sulphated glycosaminoglycans normalised to dsDNA) was highest with miR-30b-5p + miR-193b-3p, consistent with the histology results. CONCLUSION Brief ex vivo priming of MSCs with LNA-ASOs targeting anti-chondrogenic miRNAs promoted a hyaline-like programme and strengthened matrix deposition. Notably, the miR-30b-5p/miR-193b-3p pairing showed the most favourable profile, indicating promise for MSC-based strategies in cartilage regeneration. |
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