Fontiveros-Palomino M, Álvarez-Iglesias I, González-González A, Alfonso-Fernández A, Pérez-Campo FM. Modulation of osteoarthritis-related microRNAs using locked nucleic acid-antisense oligonucleotides boosts chondrogenic lineage commitment of mesenchymal progenitors. World J Stem Cells 2026; 18(4): 117652 [DOI: 10.4252/wjsc.v18.i4.117652]
Corresponding Author of This Article
Flor María Pérez-Campo, PhD, Assistant Professor, Senior Researcher, Department of Molecular Biology, Instituto de Investigación Sanitaria Marqués de Valdecilla, Faculty of Medicine, University of Cantabria, Avda Cardenal Herrera Oria S/N, Santander 39011, Cantabria, Spain. f.perezcampo@unican.es
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Apr 26, 2026 (publication date) through Apr 22, 2026
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World Journal of Stem Cells
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Fontiveros-Palomino M, Álvarez-Iglesias I, González-González A, Alfonso-Fernández A, Pérez-Campo FM. Modulation of osteoarthritis-related microRNAs using locked nucleic acid-antisense oligonucleotides boosts chondrogenic lineage commitment of mesenchymal progenitors. World J Stem Cells 2026; 18(4): 117652 [DOI: 10.4252/wjsc.v18.i4.117652]
World J Stem Cells. Apr 26, 2026; 18(4): 117652 Published online Apr 26, 2026. doi: 10.4252/wjsc.v18.i4.117652
Modulation of osteoarthritis-related microRNAs using locked nucleic acid-antisense oligonucleotides boosts chondrogenic lineage commitment of mesenchymal progenitors
Marina Fontiveros-Palomino, Itziar Álvarez-Iglesias, Alberto González-González, Ana Alfonso-Fernández, Flor María Pérez-Campo
Marina Fontiveros-Palomino, Department of Molecular Biology, Universidad de Cantabria-IDIVAL, Santander 39011, Cantabria, Spain
Itziar Álvarez-Iglesias, Alberto González-González, Flor María Pérez-Campo, Department of Molecular Biology, Instituto de Investigación Sanitaria Marqués de Valdecilla (IDIVAL), Faculty of Medicine, University of Cantabria, Santander 39011, Cantabria, Spain
Ana Alfonso-Fernández, Department of Traumatology, Hospital Universitario Marqués de Valdecilla, Santander 39008, Cantabria, Spain
Author contributions: Fontiveros-Palomino M, Álvarez-Iglesias I, González-González A, and Pérez-Campo FM contributed to manuscript preparation; Alfonso-Fernández A and Pérez-Campo FM contributed to financial support and study conceptualization; Fontiveros-Palomino M and González-González A contributed to molecular laboratory work and histology analysis; Álvarez-Iglesias I contributed to laboratory work.
Supported by a NEXT-VAL grant from the Instituto de Investigación Sanitaria Marqués de Valdecilla-IDIVAL, No. NVAL23/06; and a grant from the Instituto de Investigación Marqués de Valdecilla-IDIVAL, No. PREVAL 20/01 (to González-González A).
Institutional animal care and use committee statement: This study was approved by the Consejería de Agricultura y Ganadería de Cantabria (Spain) (Reference PI-08-21). All animal procedures were conducted in accordance with the relevant national and European regulations.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: The data supporting the findings of this study are available from the corresponding author upon reasonable request.
Corresponding author: Flor María Pérez-Campo, PhD, Assistant Professor, Senior Researcher, Department of Molecular Biology, Instituto de Investigación Sanitaria Marqués de Valdecilla, Faculty of Medicine, University of Cantabria, Avda Cardenal Herrera Oria S/N, Santander 39011, Cantabria, Spain. f.perezcampo@unican.es
Received: December 15, 2025 Revised: February 10, 2026 Accepted: March 16, 2026 Published online: April 26, 2026 Processing time: 128 Days and 23.9 Hours
Abstract
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.
Core Tip: Our study demonstrates that a brief ex vivo priming of mesenchymal stem cells (MSCs) with locked nucleic acid-antisense oligonucleotides targeting osteoarthritis-related microRNAs improved early chondrogenic programming. In rat MSC micromasses, the miR-30b-5p/miR-193b-3p combination produced higher Sox9 and Acan, lower Runx2, and stronger histological evidence of cartilage-like matrix than single inhibitors and controls. This transient, non-integrative strategy provides molecular fine-tuning of MSCs and has the potential to enhance the consistency and durability of MSC-based therapies for cartilage repair.
